WO2015024992A2 - Treatment of asthma - Google Patents

Abstract

The application is related to compositions and methods for the treatment of asthma.

Description

TREATMENT OF ASTHMA

FIELD OF THE INVENTION

This invention relates to the treatment of asthma. BACKGROUND OF THE INVENTION

Asthma is a chronic inflammatory disorder of the airways. The chronic inflammation is associated with airway hyper-responsiveness leading to recurrent episodes of wheezing, coughing, breathlessness and chest tightness, which episodes and attacks occur particularly at night or in the early morning. Asthma can be classified by severity based on the level of symptoms, airflow limitation and lung function variability. However, asthma severity may change over time, and depends not only on the severity of the underlying disease but also its responsiveness to treatment. As a consequence, asthma severity is classified on the basis of the intensity of treatment required to achieve asthma control.

Therefore, asthma control refers to the control of the manifestations of the disease and achieving and maintaining asthma control is an important treatment goal in the management of asthma. An asthmatic patient, when uncontrolled has an elevated risk of experiencing exacerbations which can be fatal. In addition, the patient experiences severe limitations in daily life. According to the worldwide accepted guidelines of "GINA" (Global Initiative for Asthma; Global Strategy for Asthma Management and Prevention (2012 update); available at htt ://www. ginasthma.org) as well as to the guidelines issued by the NIH (National Asthma Education and Prevention Program Expert Panel Report 3: Guidelines for the Diagnosis and Management of Asthma, NIH Publ. No. 08-5846, October 2007), treatment should be adjusted in a continuous cycle governed, if necessary, based on the patients' asthma control status. If asthma is not controlled on the current treatment regimen, treatment should be reduced, i.e. the controller treatment should be intensified, until control is achieved. When control can be maintained for an extended period of time, treatment intensity can be stepped down. Controller treatments are medications taken regularly on a long-term basis. They include inhaled glucocorticosteroids (ICSs) and systemic glucocorticosteroids, leukotriene modifiers, long-acting inhaled p₂-agonists (LABAs) in combination with ICSs, sustained-release theophylline, cromones, and anti-lgE.

The assessment of asthma control includes control of the clinical manifestations such as symptoms, night waking, activity limitation as well as measurements of lung function and the like. The treatment decision as well as the decision to amend the treatment, i.e. to step up or to step down intensity, is thus based on a combination of objective and subjective parameters, including lung function, frequency of use of reliever medication and asthma control assessed by asthma control questionnaire or electronic diaries. The complexity of the disease bears significant risks of both over- or undertreating patients. Undertreating patients results in an elevated risk of exacerbations with risks to patients and significant costs to the healthcare system. Overtreating patients with corticosteroids bears substantial long-term health risks for patients, particularly for children. Specifically, ICSs do have upon prolonged usage significant side effects such as loss of appetite, change in mood, osteoporosis and the like.

Therefore, an asthma assessment based on a few or even a single objective parameter would improve treatment decisions, by helping reducing over, or under treatment of patients. In particular, it would allow to identify whether a patient sufficiently benefits from the current controller treatment or not, or whether said patient should be stepped up in treatment intensity by increasing dose of the used controller treatment, or adding new controller treatments on top of, or even instead the controller treatment in use.

To date no such single selection marker has been identified that would allow to identify the patients who may benefit from a controller treatment or should be stepped up or should receive an additional treatment from which they benefit.

Analysis of the counts of eosinophil cells in peripheral blood is occasionally performed as a pharmacodynamic marker to monitor treatment effect in asthmatic patients. Such an analysis is typically done by complete blood count (CBC) and is a routine and standardized test in medical institutions and clinical practices, readily available and inexpensive.

SUMMARY OF THE INVENTION

The present invention uses the counts of eosinophil cells in peripheral blood to identify patients who can benefit from a change in different controller treatment, by increasing or decreasing intensity of the controller treatment in use, or from adding a new controller therapy in addition to the one already in use.

So far peripheral blood eosinophil counts have only been used at high levels to identify a small subset of approx. 5% of all asthma patients with severe eosinophilic asthma and recurrent life-threatening asthma exacerbations who could benefit from treatment with monoclonal antibodies against IL-5 (Pavord, I. D., et al, Lancet 380:651-659 (2012)). Contrary to severe eosinophilic asthma, the current invention uses low levels of eosinophil counts in peripheral blood, that up to now have been considered of no clinical consequence. In a particular embodiment of the invention patients are selected for treatment with a novel controller treatment based on a toll-like receptor 9 agonist packaged into virus-like particles. This novel controller treatment has recently been described showing clinical efficacy in patients with mild to moderate persistent allergic asthma (Beeh, K.-M., et al, J Allergy Clin Immunol 131 :866-874 (2013)). The patients were controlled on ICSs alone at the beginning of the trial and were subjected to a defined steroid withdrawal period in the course of the study. The clinical efficacy was hereby determined by a combination of patient-reported outcome measures such as day- and nighttime asthma symptoms, salbutamol usage, and 7-item-Asthma Control Questionnaire scores as well as objective clinical outcome measures like FEVi, fraction of exhaled nitric oxide, and blood eosinophils.

Contrary to the teaching in the art, determination of eosinophilic cell counts in peripheral blood, a readily and objectively measurable parameter, can reliably indicate whether a patient on a controller treatment would benefit from said controller treatment, or would benefit from intensifying treatment by stepping up dosing intensity of said controller treatment, or from a different controller treatment, or adding an additional controller treatment on top of said controller treatment. Such novel controller treatment can be based on a toll-like receptor 9 agonist packaged into virus-like particles recently described by Beeh. We have found that such a selection can be made based on an unexpected low level threshold of eosinophilic cell count in peripheral blood, below the level of eosinophilic cell counts considered to be clinically relevant in current clinical practice.

Thus, in a first aspect, the invention provides a composition for use in a method for treating asthma of a patient in need thereof, wherein said method comprises (i) selecting a patient on controller treatment, wherein said patient is controlled by said patient's controller treatment, and wherein said patient has an eosinophilic cell count of greater than 0.1 cells/nl in peripheral blood; and (ii) administering an effective amount of said composition to said patient, wherein said composition is selected from (a) inhaled glucocorticosteriods (ICS); (b) long-acting β₂- agonists (LABA); (c) an immune -modulating substance; or (d) any mixtures of at least one of (a), (b) and/or (c); wherein said administration is effective to improve asthma control of said patient.

The selection of the patients based on their level of eosinophilic cell count in peripheral blood allows to readily identify which patients benefit from their current controller treatment and which one's do not. Patients on controller treatment with an eosinophilic cell count in peripheral blood of greater than 0.1 cells/nl, and preferably of greater than 0.1 cells/nl and equal to or less than 0.4 cells/nl in peripheral blood, and again further preferably of greater than 0.1 cells/nl and less than 0.3 cells /nl, benefit from their current treatment and are receptive to treatments on top of their current controller treatment.

A further aspect of the invention is a pharmaceutical composition for use in a method for treating asthma of a patient in need thereof, said pharmaceutical composition comprising, essentially consisting of, or consisting of: (a) a composition of the invention; and (b) a pharmaceutically acceptable carrier.

A further aspect of the invention is a method of treating asthma of a patient in need thereof, wherein said method comprises (i) selecting a patient on controller treatment, wherein said patient is controlled by said patient's controller treatment, and wherein said patient having an eosinophilic cell count of more than 0.1 cells/nl in peripheral blood; and (ii) administering an effective amount of said composition to said patient, wherein said composition comprises, preferably consists of, (a) an inhaled glucocorticosteriod; (b) a long-acting p₂-agonist;(c) an immune-modulating substance; or (d) any mixture of at least one of (a), (b) and/or (c) wherein said administration is effective to improve asthma control of said patient.

DETAILED DESCRIPTION OF THE INVENTION

Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this invention belongs.

"allergic asthma": As used herein, the term "treating allergic asthma of a patient" refers to treating a patient who has been diagnosed as having asthma and tests positive for at least one aeroallergen in a skin prick test (SPT) or radioallergosorbent test (RAST), preferably in a skin prick test. The SPT or RAST is performed according to standard procedures known by the skilled person. A positive SPT is defined as a mean wheal diameter for the aeroallergen >3 mm and for the negative control <2 mm. The RAST is considered positive according to the manufacturer's or local laboratory definition, typically and preferably at an aeroallergen-specific IgE level >35 kU/L (Siles, R. I. and Hsieh, F. H., Cleve Clin J Med. 78(9):585-592 (2011).

"asthma control": As used herein, the term "to improve asthma control" refers to the improvement of the patient's level of asthma control due to said administering said effective amount of said composition to said patient as compared to the patient's level of asthma control due to the patient's controller treatment alone. Preferably, the level of the patient's asthma control is determined by assessment of the patient's clinical manifestations of asthma or inflammatory or patho-physiological parameters comprising (i) assessment of the number and frequency of occurrence of patient's daytime symptoms such as breathlessness, wheezing, cough, or chest tightness; (ii) assessment of the number and frequency of patient's limitation of activities; (iii) assessment of the number and frequency of patient's nocturnal symptoms/awakening; (iv) assessment of the number and frequency of patient's need for reliever/rescue treatment; (v) determination of patient's lung function as expressed by the measurement of the peak expiratory flow (PEF); (vi) determination of patient's lung function as expressed by the measurement of the forced expiratory volume in 1 second (FEVi); (vii) determination of patient's airway responsiveness as expressed by the PC20 value upon airway challenges with inhaled metacholine or histamine; (viii) determination of patient's eosinophilic cell count in peripheral blood; (ix) assessment of the number and frequency of patient's exacerbations in the past year; (x) assessment by way of any asthma control questionnaires or diaries such as ACQ, ACQ-7, MiniAQLQ, ACT, C-ACT, ATAQ; and (xi) any patient's clinical manifestation of asthma or inflammatory or patho-physiological parameter or any other measure as referred to in the guidelines of GINA (Global Initiative for Asthma; Global Strategy for Asthma Management and Prevention (2012 update); available at http://www.ginasthma.org) or in the guidelines of NIH (National Asthma Education and Prevention Program Expert Panel Report 3: Guidelines for the Diagnosis and Management of Asthma, NIH Publ. No. 08-5846, October 2007); wherein said improvement of asthma control refers to the improvement of at least one of said patient's clinical manifestation of asthma or inflammatory or pathophysiological parameter or any other measure identified in any one of (i) to (xi) due to said administering of said effective amount of said composition to said patient as compared to said patient's clinical manifestation of asthma or inflammatory or patho-physiological parameter or any other measure due to the patient's controller treatment alone.

Several standardized measures for assessing patient's level of asthma control have been developed, which provide numerical values to distinguish different levels of control. Examples of validated instruments include the 7-item Asthma Control Questionnaire (ACQ-7) (Juniper, E. F., et al. Eur Respir J 14:32-38 (1999), the Mini Asthma Quality of Life Questionnaire (MiniAQLQ) (Wilson, S. R., et al, J Allergy Clin Immunol 129:S88-123 (2012), the Asthma Control Test (ACT) (Nathan, R.A., et al, J Allergy Clin Immunol 113:59-65 (2004), the Childhood Asthma Control Test (C-ACT) (Uu, A. H., et al, J Allergy Clin Immunol 119:817-25 (2007), the Asthma Therapy Assessment Questionnaire (ATAQ) (Vollmer, W. M., et al, Am J Respir Crit Care Med 160: 1647-52 (1999), the Asthma Control Scoring System (Boulet, L. P., et al, Chest 122:2217-2223 (2002).

Further preferably the term "to improve asthma control" refers to the improvement of the patient's level of asthma control due to said administering said effective amount of said composition to said patient as compared to the patient's level of asthma control due to the patient's controller treatment alone, wherein said level of the patient's asthma control is determined by assessment of the patient's ACQ-7, preferably wherein said level of the patient's asthma control is solely determined by assessment of the patient's ACQ-7. Again further preferably, the term "to improve asthma control" refers to the improvement of the patient's ACQ-7 score due to said administering said effective amount of said composition to said patient as compared to the patient's ACQ-7 score due to the patient's controller treatment alone, wherein said improvement of said patient's ACQ-7 score consists of at least 0.2, preferably consists of at least 0.3, more preferably consists of at least 0.4, and again further preferably consists of at least 0.5. Again still further preferably, the term "to improve asthma control" refers to the improvement of the patient's ACQ-7 score due to said administering said effective amount of said composition to said patient as compared to the patient's ACQ-7 score due to the patient's controller treatment alone, wherein said improvement of said patient's ACQ-7 score occurs after at least 12 weeks, preferably after at least 8 weeks, more preferably after at least 4 weeks after a first administration of said effective amount of said composition.

"chemoattractant receptor-homologous molecule expressed on Th2 cells antagonist

(CRTh2 antagonist)"; CRTh2 anatagonists are well known immune-modulating substances. The CRTh2 receptor is expressed on Th2-cells, basophils and eosinophils, and its ligand, prostaglandin D2 (PGD2), is released by mast cells. The PGD2/CRTh2 axis plays a key role in the migration and activation of inflammatory cells leading to many symptoms of asthma including coughing, difficulty breathing and exacerbations. Hence, prevention of CRTh2 signalling may allow enhancing disease control. CRTh2 antagonists include substances like ARRY005, Vidupiprant, ADC-3680, OC000459.

"chemokine antagonist"; Chemokine antagonists are well known immune-modulating substances. They block chemokine receptors or neutralize chemokines. Chemokine receptors include CCR1, CCR3 CCR4, and CCR5. Substances which may prevent chemokine signalling include Mogalizumab (CCR4 antagonist), GW-824575 (CCR3 antagonist), Bertilimumab (CCR3-antagonist), CT-2008 (CCR5 antagonist) and NI-0701 which targets CCL5, a chemokine from the C-C subfamily that binds to cell receptor CCR1, CCR3 and CCR5.

"controlled": As used herein, the term "controlled" refers to a patient having an ACQ-7 score of < 1.5. The 7-item Asthma Control Questionnaire ACQ-7 is a validated composite score based on the average of 7 items each scored 0-6, including patient's scores of 5 disease-related items (woken at night by asthma, awake in the morning with symptoms, limitation of daily activities, shortness of breath, wheezing) + 1 score for number of puffs of short acting β₂- agonists (SABA) + 1 score for FEVi predicted (Juniper, E. F., et al. Eur Respir J 14:32-38 (1999)). Improvement of said controlled patient may lead to a "well-controlled" patient defined as having an ACQ-7 score < 0.75 (Juniper, E. F., et al, Respir Med 100:616-621 (2006).

"controller treatment": As used herein, the term "controller treatment" refers to medications taken regularly, typically and preferably daily, by a patient, typically on a long-term basis, including inhaled glucocorticosteroids (ICSs) and and systemic glucocorticosteroids, leukotriene modifiers, long-acting inhaled p₂-agonists (LABAs) in combination with ICSs, sustained-release theophylline, cromones, and anti-lgE. As used herein, the term "controller treatment", typically and preferably, excludes reliever medication as defined herein. Thus, controller treatment and reliever medication, in the context of the present invention, are, typically and preferably, two different and distinguished treatments. The aforementioned controller treatments are known by the skilled in the art and are recommended and described by the worldwide accepted guidelines of GINA (Global Initiative for Asthma; Global Strategy for Asthma Management and Prevention (2012 update); available at http://www.ginasthma.org) or the guidelines of NIH (National Asthma Education and Prevention Program Expert Panel Report 3: Guidelines for the Diagnosis and Management of Asthma, NIH Publ. No. 08-5846, October 2007). Preferably, the term "controller treatment", as used herein refers to treatments by way of inhaled glucocorticosteroids (ICSs), leukotriene modifiers, or long-acting inhaled ₂-agonists (LABAs) in combination with ICSs or sustained-release theophylline, all treatments taken daily by the patient, typically on a long-term basis. Alternatively preferably, the term "controller treatment", as used herein refers to treatments by way of inhaled glucocorticosteroids (ICSs) or long-acting inhaled ₂-agonists (LABAs) in combination with ICSs, both treatments taken daily by the patient, typically on a long-term basis.

"eosinophilic cell count in peripheral blood": As used herein, the term "eosinophilic cell count in peripheral blood" refers to a value of blood eosinophils typically and preferably analyzed by automated CBC (complete blood count). CBC with automated analyzers is a well- validated and routine method for assessing the number of leukocytes in the blood and is well known to the skilled in the art. For this purpose, venous blood is drawn and put into a tube typically and preferably containing EDTA. The CBC is then determined using an automated analyzer, such as the Medonic M-Series, Beckman Coulter LH series, Roche Sysmex XE-2100, Siemens ADVIA 120 and 2120, Abbott CELL-DYN series, Mindray BC series or the like. The total number of white blood cells is multiplied by the percentage of eosinophils to provide said "eosinophilic cell count in peripheral blood". The value is typically provided by number of eosinophil cell counts/nl with one decimal. In a preferred manner of the present invention, said analysis of said eosinophilic cell count in peripheral blood is conducted by a state of the art cell counter, typically and preferably by the automated analyzer Roche Sysmex XE-2100 or Beckman Coulter L750, typically and preferably using a laser based flow cytometer as analytical module, wherein said number of eosinophil cell counts in peripheral blood, i.e. the total number of eosinophil cells in peripheral blood, is provided in counts/nl (10^~9 liter), wherein said number is rounded to the first figure after the decimal. Typically and preferably, the rounding is governed as follows: if the second figure after the decimal is a "0", "1", "2", "3" or "4", then it has to be down rounded, wherein if the second figure after the decimal is a "5", "6", "7", "8" or "9", then it has to be up rounded. For example, a value of 0.15 would be rounded up to 0.2, wherein a value of 0.24 would be rounded down to 0.2. Again in a more preferred manner, said analysis of said eosinophilic cell count in peripheral blood is conducted by the automated analyzer Roche Sysmex XE-2100, typically and preferably using a laser based flow cytometer as analytical module, wherein said number of eosinophil cell counts in peripheral blood, i.e. the total number of eosinophil cells in peripheral blood, is provided in counts/nl (10^~9 liter), wherein said number is rounded to the first figure after the decimal. Typically and preferably, the rounding is governed as follows: if the second figure after the decimal is a "0", "1", "2", "3" or "4", then it is down rounded, wherein if the second figure after the decimal is a "5", "6", "7", "8" or "9", then it is up rounded. For example, a value of 0.15 is rounded up to 0.2, wherein a value of 0.24 is rounded down to 0.2.

"GINA Step 2": GINA Steps are controller treatments recommended by the worldwide accepted guidelines "GINA" (Global Initiative for Asthma; Global Strategy for Asthma Management and Prevention (2012 update); available at http://www.ginasthma.org) as alternatives for controlling asthma. As used herein, the term "GINA Step 2" refers to a defined selection of controller treatments as described in Figure 4.3-2. on page 67 of said 2012 update, wherein said defined selection of controller treatments are termed therein "Controller options".

"GINA Step 3": GINA Steps are controller treatments recommended by the worldwide accepted guidelines "GINA" (Global Initiative for Asthma; Global Strategy for Asthma Management and Prevention (2012 update); available at http://www.ginastfa.ma.ofg) as alternatives for controlling asthma. As used herein, the term "GINA Step 3" refers to a defined selection of controller treatments as described in Figure 4.3-2. on page 67 of said 2012 update, wherein said defined selection of controller treatments are termed therein "Controller options".

"GINA Step 4": GINA Steps are controller treatments recommended by the worldwide accepted guidelines "GINA" (Global Initiative for Asthma; Global Strategy for Asthma Management and Prevention (2012 update); available at http://www.ginasthma.org) as alternatives for controlling asthma. As used herein, the term "GINA Step 4" refers to a defined selection of controller treatments as described in Figure 4.3-2. on page 67 of said 2012 update, wherein said defined selection of controller treatments are termed therein "Controller options". "immune-modulating substance": As used herein, the term "immune-modulating substance" refers to a substance which (a) is capable of inducing modulating and/or enhancing an effect on a cell of the immune system of a mammalian, preferably of a human, upon exposure of said cell or said animal, or preferably said human, to said immune-modulating substance, as compared to a suitable control. This observation can relate to any parameter known in the art to be indicative for a cell of the immune system of a mammalian, preferably of a human. As used herein, the term "immune -modulating substance" further refers to a substance which (b) is capable of neutralizing or binding a substance produced by a cell of the immune system of a mammalian, preferably of a human; provided however that said immune-modulating substance capable of neutralizing or binding a substance produced by a cell of the immune system of a mammalian, preferably of a human is, preferably, not a monoclonal antibody against IL-4, IL-5 and/or IL-13 and/or or against the alpha subunit of the interleukin-4 receptor.

Cells of the immune system include B-cells, T-cells, plasma cells, dendritic cells (e.g myeloid dendritic cells, plasmacytoid dendritic cells and Langerhans cells), macrophages, monocytes, eosinophils, neutrophils, basophils and mast cells. Substances produced by immune cells can be classified into secreted cytokines, chemokines and cell surface receptors.

Preferably, the term "immune-modulating substance", as used herein, refers to a substance which (a) is capable of inducing modulating and/or enhancing an effect on a cell of the immune system of a mammalian, preferably of a human, upon exposure of said cell or said animal, or preferably said human, to said immune -modulating substance, as compared to a suitable control.

"Immune-modulating nucleic acid (INA)": As used herein, the term "immune- modulating nucleic acid" refers to a nucleic acid capable of inducing, modulating and/or enhancing an immune response, wherein "said inducing, modulating and/or enhancing an immune response" refers to said INA being capable of inducing, modulating or enhancing the immune response of a cell or an animal, preferably a human, upon exposure of said cell or said animal, or preferably of said human, to said INA, as compared to a suitable control. This observation can relate to any parameter known in the art to be indicative for an immune response, preferably to the formation of cytokines or to cytotoxicity. For example, the formation of cytokines such as 11-12 can be assessed as described in Example 1 1 of WO2007/068747. The lytic activity of cytotoxic T cells can be, for example, measured, e.g. using a ⁵¹Cr release assay, with and without the INA. The amount of the substance at which the CTL lytic activity is enhanced as compared to the CTL lytic activity without the INA is said to be an amount sufficient to enhance the immune response.

INAs, as used herein, comprise ribonucleic acids and, in particular, desoxyribonucleic acids, wherein both, ribonucleic acids and desoxyribonucleic acids may be either double stranded or single stranded. Preferred INAs are desoxyribonucleic acids, wherein further preferably said desoxyribonucleic acids are single stranded. Preferably, INA contain at least one CpG motif comprising an unmethylated C. Very preferred INAs comprise at least one CpG motif, wherein said at least one CpG motif comprises or preferably consist of at least one, preferably two, CG dinucleotide, wherein the C is unmethylated. Preferably, but not necessarily, said CG dinucleotide is part of a palindromic sequence. The term "immune-modulating nucleic acid" as used herein also refers to nucleic acids that contain modified bases, preferably 4-bromo- cytosine. Specifically preferred in the context of the invention are INAs which are capable of stimulating IFN-alpha production in dendritic cells.

"oligonucleotide": As used herein, the term "oligonucleotide" refers to a nucleic acid sequence comprising 2 or more nucleotides, preferably at least about 6 nucleotides to about 100,000 nucleotides, more preferably about 6 to about 2000 nucleotides, and still more preferably about 6 to about 300 nucleotides, even more preferably about 20 to about 300 nucleotides, and even more preferably about 20 to about 100 nucleotides, and most preferably 20 to 40 nucleotides. Very preferably oligonucleotides comprise about 30 nucleotides, more preferably oligonucleotides comprise exactly 30 nucleotides, and most preferably oligonucleotides consist of exactly 30 nucleotides. The term oligonucleotide also refers to a nucleic acid comprising more than 100 to about 2000 nucleotides, preferably more than 100 to about 1000 nucleotides, and more preferably more than 100 to about 500 nucleotides.

Oligonucleotides are polyribonucleotides or polydeoxyribonucleotides and are preferably selected from (a) unmodified RNA or DNA, and (b) modified R A or DNA. The modification may comprise the backbone or nucleotide analogues. Oligonucleotides are preferably selected from (a) single- and double-stranded DNA, (b) DNA that is a mixture of single- and double- stranded regions, (c) single- and double-stranded RNA, (d) RNA that is mixture of single- and double-stranded regions, or (e) hybrid molecules comprising DNA and RNA that are single- stranded or, more preferably, double-stranded or a mixture of single- and double-stranded regions. In a further embodiment oligonucleotides are triple-stranded regions and higher-ordered structures comprising RNA or DNA or both RNA and DNA. In further embodiments, oligonucleotides are synthetic, genomic or recombinant. Double-stranded RNA, for example, poly (I:C) are also known and typically activate TLR3 (Alexopoulou et al., Nature 413:732-738 (2001). In one embodiment, said INA is a double stranded ribonucleic acid, wherein said double stranded ribonucleic acid is poly(LC) or a derivative thereof. In one embodiment oligonucleotide refers to (a) DNA or RNA containing at least one modified nucleotide or at least one nucleotide analogue, or (b) to DNA or RNA with backbones modified for stability or for other reasons. Preferred nucleotide modifications/analogs are selected from (a) peptide nucleic acid, (b) inosin, - l i ^¬ fe) tritylated bases, (d) phosphorothioates, (e) alkylphosphorothioates, (f) 5-nitroindole desoxyribofuranosyl, (g) 5-methyldesoxycytosine, or (h) 5,6-dihydro-5,6- dihydroxydesoxythymidine. Phosphothioated nucleotides are protected against degradation, in particular by nucleases, in a cell or an organism and are therefore preferred nucleotide modifications. Further preferred are chemically, enzymatically or metabolically modified forms of polynucleotides as typically found in nature, as well as the chemical forms of DNA and RNA characteristic of viruses and cells. Other nucleotide analogs or modifications will be evident to those skilled in the art. However, unmodified oligonucleotides consisting exclusively of phosphodiester bound nucleotides, typically are more active as INA than modified nucleotides and are therefore generally preferred in the context of the invention. Most preferred are oligonucleotides consisting exclusively of phosphodiester bound deoxinucleotides. Further preferred are oligonucleotides capable of stimulating IFN-alpha production in cells, preferably in dendritic cells. Very preferred oligonucleotides capable of stimulating IFN-alpha production in cells are selected from A-type CpGs and C-type CpGs, again further prerably A-type CpGs.

"CpG motif: As used herein, the term "CpG motif either refers to a CG dinucleotide, wherein the C is unmethylated or to a pattern of nucleotides that includes or consists of such an unmethylated central CpG, i.e. the unmethylated CpG dinucleotide, surrounded by at least one base, preferably one or two nucleotides, flanking (on the 3' and the 5' side of) the central CpG. Typically and preferably, the CpG motif as used herein, comprises or alternatively consists of the unmethylated CpG dinucleotide and two nucleotides on its 5' and 3' ends.

"CpG" / "unmethylated CpG-containing oligonucleotide": As used herein, the term "unmethylated CpG-containing oligonucleotide" or "CpG" refers to an oligonucleotide, preferably to an oligodeoxynucleotide, containing at least one CpG motif. Thus, a CpG contains at least one unmethylated cytosine, guanine dinucleotide. Preferred CpGs stimulate/activate, e.g. have a mitogenic effect on, or induce or increase cytokine expression by, a vertebrate bone marrow derived cell. For example, CpGs can be useful in activating B cells, NK cells and antigen-presenting cells, such as dendritic cells, monocytes and macrophages. Preferably, CpG relates to an oligodeoxynucleotide, preferably to a single stranded oligodeoxynucleotide, containing an unmethylated cytosine followed 3' by a guanosine, wherein said unmethylated cytosine and said guanosine are linked by a phosphate bond, wherein preferably said phosphate bound is a phosphodiester bound or a phosphothioate bound, and wherein further preferably said phosphate bond is a phosphodiester bound. CpGs can include nucleotide analogs such as analogs containing phosphorothioester bonds and can be double-stranded or single-stranded. Generally, double-stranded molecules are more stable in vivo, while single-stranded molecules typically have increased immune activity. Preferably, as used herein, a CpG is an oligonucleotide that is at least about ten nucleotides in length and comprises at least one CpG motif, wherein further preferably said CpG is 10 to 60, more preferably 15 to 50, still more preferably 20 to 40, still more preferably about 30, and most preferably exactly 30 nucleotides in length. A CpG may consist of methylated and/or unmethylated nucleotides, wherein said at least one CpG motif comprises at least one CG dinucleotide wherein the C is unmethylated. The CpG may also comprise methylated and unmethylated sequence stretches, wherein said at least one CpG motif comprises at least one CG dinucleotide wherein the C is unmethylated. Very preferred CpGs consist exclusively of unmethylated nucleotides. Very preferably, CpG relates to a single stranded oligodeoxynucleotide containing an unmethylated cytosine followed 3' by a guanosine, wherein said unmethylated cytosine and said guanosine are linked by a phosphodiester bound. Still more preferably, CpG relates to a single stranded oligodeoxynucleotide containing an unmethylated cytosine followed 3' by a guanosine, wherein said unmethylated cytosine and said guanosine are linked by a phosphodiester bound, and wherein said CpG consist exclusively of unmethylated nucleotides. Most preferably, CpG relates to a single stranded oligodeoxynucleotide of about 30 nucleotides in length, containing an unmethylated cytosine followed 3' by a guanosine, wherein said unmethylated cytosine and said guanosine are linked by a phosphodiester bound, and wherein said CpG consist exclusively of unmethylated nucleotides.

The CpGs can include nucleotide analogs such as analogs containing phosphorothioester bonds and can be double-stranded or single-stranded. Generally, phosphodiester CpGs are A- type CpGs as indicated below, while phosphothioester stabilized CpGs are B-type CpGs or C- type CpGs. Preferred CpG oligonucleotides in the context of the invention are A-type CpGs and C-type CpG, most preferred are A-type CpGs.

"A-type CpG": As used herein, the term "A-type CpG" refers to an oligodeoxynucleotide (ODN) comprising at least one CpG motif. A-type CpGs preferentially stimulate activation of T cells and the maturation of dendritic cells and are capable of stimulating IFN-alpha production. In A-type CpGs, the nucleotides of the at least one CpG motif are linked by at least one phosphodiester bond. A-type CpGs comprise at least one phosphodiester bond CpG motif which may be flanked at its 5' end and/or, preferably and, at its 3' end by phosphorothioate bound nucleotides. Preferably, the CpG motif, and hereby preferably the CG dinucleotide and its immediate flanking regions comprising at least one, preferably two nucleotides, are composed of phosphodiester nucleotides. Preferred A-type CpGs exclusively consist of phosphodiester (PO) bond nucleotides. Further preferred A-type CpGs do not comprise phosphothioate bounds. Typically and preferably, the term "A-type CpG" or "D-type CpG" as used within this specification, refers to an oligodesoxynucleotide (ODN) comprising at least one CpG motif and having poly G motifs at the 5' and/or 3' ends. Typically and preferably, the poly G motif comprises or alternatively consists of at least one, preferably at least three, at least 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 Gs (guanosines), most preferably by at least 10 Gs. In some embodiments, the 5 ' and/or 3 ' ends, typically and preferably at least one G of the poly G motifs at the 5' and/or 3' ends, preferably at least two, three or four, even more preferably all Gs of the poly G motif, are phoshorothioate modified. However, in a very preferred embodiment, all Gs of the poly G motif are linked by phosphodiester bonds. Preferably, the A-type CpG of the invention comprises or alternatively consists of a palindromic sequence. Typically and preferably, the CpG motif is part of said palindromic sequence. Typically and preferably, all nucleotides, but at least the CpG motif of the palindromic sequence, are composed of phosphodiester nucleotides. Typically and preferably, the palindromic sequence is SEQ ID NO: l . Very preferred A-type CpGs are 16 to 30 nucleotides in length, consist exclusively of phosphodiester bound nucleotides, comprise a palindromic sequence, preferably the palindromic sequence of SEQ ID NO: l, and are flanked at their 5' and at their 3' end by a poly G motif consisting of 3 to 10 Gs.

"B-type CpG": As used herein, the term "B-type CpG" relates to a CpG oligonucleotide which predominantly or preferably exclusively consists of modified nucleotides, preferably phosphorothioate modified nucleotides. B-type CpGs stimulate preferentially B-cell and to some extent NK-cell activation and cytokine production.

"C-type CpG": As used herein, the term "C-type CpG" relates to a CpG oligonucleotide which like a B-type oligonucleotide predominantly or preferably exclusively consists of modified nucleotides, preferably phosphorothioate modified nucleotides. Examples of C-type CpGs have been described in WO2005/042018A2 and in Vollmer et al. 2004, Eur. J. Immunol. 43:351-262 which are incorporated herein by reference. Specific reference is made to SEQ IDs NO: 1 to 69 of WO2005/042018A2. C-type CpGs combine effects of A-type and B-type CpGs and stimulate B-cell or NK-cell activation and IFN-alpha production, preferably IFN-alpha production in dendritic cells. C-type CpCs which are capable of stimulating IFN-alpha production, preferably in dendritic cells, are generally preferred in the context of the invention. In contrast to A-type CpGs, C-type CpGs do not comprise poly-G stretches.

"palindromic sequence": A palindromic sequence is a nucleotide sequence which, when existing in the form of a double stranded nucleic acid with regular base pairing (A/T; C/G), would consist of two single strands with identical sequence in 5 '-3' direction. An INA of the present invention preferably comprises a palindromic sequence, preferably a palindromic sequence consisting of at least 6, preferably of at least 7, 8, 9 or 10, most preferably of exactly 10 nucleotides, wherein most preferably said palindromic sequence preferably comprises at least one, preferably two, CpG motif. Palindromic sequences of INAs useful in the context of the present invention are, for example, described in Yamamoto et al. 1992, J. Immunol. 148(12):4072-4076 and Kuramoto et al. 1992, Jpn. J. Cancer Res. 83: 1128-1131.

"inhaled glucocorticosteriod (ICS)": Inhaled glucocorticosteriods (ICSs) as well as the different ways of treatment with ICSs are known to the skilled person in the art and are described within the worldwide accepted guidelines of GINA (Global Initiative for Asthma; Global Strategy for Asthma Management and Prevention (2012 update); available at htt ://www. inasthma.org) or the guidelines of NIH (National Asthma Education and Prevention Program Expert Panel Report 3 : Guidelines for the Diagnosis and Management of Asthma, NIH Publ. No. 08-5846, October 2007). Well-known and preferred ICSs are selected from among beclomethasone, budesonide, butixocortpropionate, ciclesonide, dexamethasone, flunisolide, fluticasone, mometasone, prednisolone, prednisone, rofleponide and triamcinolone, optionally in the form of the racemates, enantiomers or diastereomers thereof and optionally in the form of the salts and derivatives thereof, the solvates and/or hydrates thereof, said salts and derivatives of said ICS typically and preferably selected from among alkali metal salts, such as for example sodium or potassium salts, sulphobenzoates, phosphates, isonicotinates, acetates, propionates, dipropionates, dihydrogen phosphates, palmitates, pivalates or furoates. More preferably, said inhaled glucocorticosteriod is selected among from beclomethasone, beclomethasone dipropionate, budesonide, ciclesonide, flunisolide, fluticasone, fluticasone propionate, mometasone, mometasone furoate, triamcinolone and triamcinolone acetonide, optionally in the form of the racemates, enantiomers or diastereomers thereof and optionally in the form of the salts and derivatives thereof, the solvates and/or hydrates thereof.

"low dose inhaled glucocorticosteriods": As used herein, the term "low dose inhaled glucocorticosteriods" is known to the skilled in the art and refers to a daily dose of an ICS as suggested and described within the worldwide accepted guidelines of GINA (Global Initiative for Asthma; Global Strategy for Asthma Management and Prevention (2012 update); available at http ://www. ginasthma.org, page 3 1 ) or any other estimated equipotent daily dose of any other ICS. Preferably, the term "low dose inhaled glucocorticosteriods" refers to a daily dose of an ICS selected among from (i) 200-500 μg of beclomethasone dipropionate (BDP) as a suspension in chlorofluorocarbons (CFC); (ii) 100-250 μg of beclomethasone dipropionate (BDP) as a solution in hydro fluoroalkanes (HFA); (iii) 200-400 μg of budenoside; (iv) 80-160 μg of ciclesonide; (v) 500-1000 μg of flunisolide; (vi) 100-250 μg of fluticasone propionate; (vii) 200 μg of mometasone furoate; (viii) 400-1000 μg of triamcinolone acetonide; or (ix) any other daily dose of any other ICS being equipotent to any one of (i) to (viii). Further preferably, the term "low dose inhaled glucocorticosteriods" refers to a daily dose of an ICS selected among from (i) 200- 500 μg of beclomethasone dipropionate (BDP) as a suspension in chlorofluorocarbons (CFC); (ii) 100-250 μg of beclomethasone dipropionate (BDP) as a solution in hydrofluoroalkanes (HFA); (iii) 200-400 μ§ of budenoside; (iv) 80-160 μ§ of ciclesonide; (v) 500-1000 μ§ of flunisolide; (vi) 100-250 μg of fluticasone propionate; (vii) 200 μg of mometasone furoate; or (viii) 400-1000 μg of triamcinolone acetonide. Again further preferably, the term "low dose inhaled glucocorticosteriods" refers to a daily dose of (i) 200-500 μg of beclomethasone dipropionate (BDP) as a suspension in chlorofluorocarbons (CFC) or (ii) 100-250 μg of fluticasone propionate or (iii) an equivalent to (i) or (ii), wherein said equivalent corresponds to an equipotent dose of an ICS corresponding to (i) or (ii) upon efficacy literature known to the skilled person in the art. And still further preferably, the term "low dose inhaled glucocorticosteriods" refers to a daily dose of 200-500 μg of beclomethasone dipropionate (BDP) as a suspension in chlorofluorocarbons (CFC) or an equivalent thereof, wherein said equivalent corresponds to an equipotent dose of an ICS corresponding to said daily dose of 200- 500 μg of beclomethasone dipropionate (BDP) as a suspension in chlorofluorocarbons (CFC) upon efficacy literature known to the skilled person in the art. Alternatively still further preferably, the term "low dose inhaled glucocorticosteriods" refers to a daily dose of 100-250 μg of fluticasone propionate or an equivalent thereof, wherein said equivalent corresponds to an equipotent dose of an ICS corresponding to said daily dose of 100-250 μg of fluticasone propionate upon efficacy literature known to the skilled person in the art.

"medium dose inhaled glucocorticosteriods": As used herein, the term "medium dose inhaled glucocorticosteriod" is known to the skilled in the art and refers to a daily dose of an ICS as suggested and described within the worldwide accepted guidelines of GINA (Global Initiative for Asthma; Global Strategy for Asthma Management and Prevention (2012 update); available at htt ://www. inasthma.org, page 31) or any other estimated equipotent daily dose of any other ICS. Preferably, the term "medium dose inhaled glucocorticosteriods" refers to a daily dose of an ICS selected among from (i) >500- 1000 μg of beclomethasone dipropionate (BDP) as a suspension in chlorofluorocarbons (CFC); (ii) >250-500 μg of beclomethasone dipropionate (BDP) as a solution in hydrofluoroalkanes (HFA); (iii) >400-800 μg of budenoside; (iv) >160- 320 μg of ciclesonide; (v) > 1000-2000 μg of flunisolide; (vi) >250-500 μg of fluticasone propionate; (vii) >400 μg of mometasone furoate; (viii) > 1000-2000 μg of triamcinolone acetonide; or (ix) any other daily dose of any other ICS being equipotent to any one of (i) to (viii). Further preferably, the term "medium dose inhaled glucocorticosteriods" refers to a daily dose of an ICS selected among from (i) >500- 1000 μg of beclomethasone dipropionate (BDP) as a suspension in chlorofluorocarbons (CFC); (ii) >250-500 μg of beclomethasone dipropionate (BDP) as a solution in hydrofluoroalkanes (HFA); (iii) >400-800 μg of budenoside; (iv) >160- 320 μg of ciclesonide; (v) > 1000-2000 μg of flunisolide; (vi) >250-500 μg of fluticasone propionate; (vii) >400 μg of mometasone furoate; or (viii) > 1000-2000 μg of triamcinolone acetonide. Again further preferably, the term "medium dose inhaled glucocorticosteriods" refers to a daily dose of (i) >500-1000 μg of beclomethasone dipropionate (BDP) as a suspension in chlorofluorocarbons (CFC) or (ii) >250-500 μg of fluticasone propionate or (iii) an equipotent dose of an ICS corresponding to (i) or (ii) upon efficacy literature known to the skilled person in the art. And still further preferably, the term "low dose inhaled glucocorticosteriods" refers to a daily dose of >500-1000 μg of beclomethasone dipropionate (BDP) as a suspension in chlorofluorocarbons (CFC) or an equivalent thereof, wherein said equivalent corresponds to an equipotent dose of an ICS corresponding to said daily dose of >500-1000 μg of beclomethasone dipropionate (BDP) as a suspension in chlorofluorocarbons (CFC) upon efficacy literature known to the skilled person in the art. Alternatively still further preferably, the term "low dose inhaled glucocorticosteriods" refers to a daily dose of >250-500 μg of fluticasone propionate or an equivalent thereof, wherein said equivalent corresponds to an equipotent dose of an ICS corresponding to said daily dose of >250-500 μg of fluticasone propionate upon efficacy literature known to the skilled person in the art.

"high dose inhaled glucocorticosteriods": As used herein, the term "high dose inhaled glucocorticosteriods" is known to the skilled in the art and refers to a daily dose of an ICS as suggested and described within the worldwide accepted guidelines of GINA (Global Initiative for Asthma; Global Strategy for Asthma Management and Prevention (2012 update); available at htt ://www. inasthma.org, page 31) or any other estimated equipotent daily dose of any other ICS. Preferably, the term "high dose inhaled glucocorticosteriods" refers to a daily dose of an ICS selected among from (i) > 1000-2000 μg of beclomethasone dipropionate (BDP) as a suspension in chlorofluorocarbons (CFC); (ii) >500-1000 μg of beclomethasone dipropionate (BDP) as a solution in hydrofluoroalkanes (HFA); (iii) >800-1600 μg of budenoside; (iv) >320- 1280 μg of ciclesonide; (v) >2000 μg of flunisolide; (vi) >500-1000 μg of fluticasone propionate; (vii) >800 μg of mometasone furoate; (viii) >2000 μg of triamcinolone acetonide; or (ix) any other daily dose of any other ICS being equipotent to any one of (i) to (viii). Further preferably, the term "high dose inhaled glucocorticosteriods" refers to a daily dose of an ICS selected among from (i) > 1000-2000 μg of beclomethasone dipropionate (BDP) as a suspension in chlorofluorocarbons (CFC); (ii) >500-1000 μg of beclomethasone dipropionate (BDP) as a solution in hydrofluoroalkanes (HFA); (iii) >800-1600 μg of budenoside; (iv) >320-1280 μg of ciclesonide; (v) >2000 μg of flunisolide; (vi) >500-1000 μg of fluticasone propionate; (vii) >800 μg of mometasone furoate; or (viii) >2000 μg of triamcinolone acetonide. Again further preferably, the term "high dose inhaled glucocorticosteriods" refers to a daily dose of (i) >1000- 2000 μg of beclomethasone dipropionate (BDP) as a suspension in chlorofluorocarbons (CFC) or (ii) >500-1000 μg of fluticasone propionate or (iii) an equipotent dose of an ICS corresponding to (i) or (ii) upon efficacy literature known to the skilled person in the art. And still further preferably, the term "low dose inhaled glucocorticosteriods" refers to a daily dose of >1000- 2000 μg of beclomethasone dipropionate (BDP) as a suspension in chlorofluorocarbons (CFC) or an equivalent thereof, wherein said equivalent corresponds to an equipotent dose of an ICS corresponding to said daily dose of > 1000-2000 μg of beclomethasone dipropionate (BDP) as a suspension in chlorofluorocarbons (CFC) upon efficacy literature known to the skilled person in the art. Alternatively still further preferably, the term "low dose inhaled glucocorticosteriods" refers to a daily dose of >500-1000 μg of fluticasone propionate or an equivalent thereof, wherein said equivalent corresponds to an equipotent dose of an ICS corresponding to said daily dose of >500-1000 μg of fluticasone propionate upon efficacy literature known to the skilled person in the art.

"long-acting p₂-agonist (LABA)": Long-acting p₂-agonists (LABAs) as well as the different ways of treatment with LABAs, in particular in combination with ICSs, are known to the skilled person in the art and are described within the worldwide accepted guidelines of GINA (Global Initiative for Asthma; Global Strategy for Asthma Management and Prevention (2012 update); available at http://www.ginasthma.org) or the guidelines of NIH (National Asthma Education and Prevention Program Expert Panel Report 3: Guidelines for the Diagnosis and Management of Asthma, NIH Publ. No. 08-5846, October 2007). Well-known and preferred LABAs are selected from among albuterol, arformoterol, bambuterol, bitolterol, broxaterol, carbuterol, clenbuterol, fenoterol, formoterol, hexoprenaline, ibuterol, isoetharine, isoprenaline, levosalbutamol, mabuterol, meluadrine, metaproterenol, orciprenaline, pirbuterol, procaterol, reproterol, TD 3327, ritodrine, salmeterol, salmefamol, soterenot, sulphonterol, tiaramide, terbutaline and tolubuterol, optionally in the form of the racemates, the enantiomers, the diastereomers and optionally the pharmacologically acceptable acid addition salts and the hydrates thereof.

"patient": As used herein, the term "patient" refers preferably to a human with an age of older than 5 years, more preferably to a human equal to or older than 6 years, again more preferably to a human equal to or older than 12 years, again more preferably to a human equal to or older than 18 years, and preferably equal to or younger than 65 years, and again more preferably with an age of between 18 and 65 years. "QPGIO": As used herein, the term "QpGlO" (or interchangeably used with "QbGlO") refers to a virus-like particle of an R A-phage Q consisting of coat proteins with the amino acid sequence of SEQ ID NO: 7 into which an immune-modulating substance is packaged, wherein said immune -modulating substance is an unmethylated CpG-containing oligonucleotide consisting of the sequence GGGGGGGGGGGACGATCGTCGGGGGGGGGG (SEQ ID NO:6), and wherein said unmethylated CpG containing oligonucleotide exclusively consists of phosphodiester connected deoxynucleotides. Preferably, as used herein, the term "QPGIO" (or interchangeably used with "QbGlO") refers to a virus-like particle of an RNA-phage Q consisting of coat proteins with the amino acid sequence of SEQ ID NO: 7 into which an unmethylated CpG-containing oligonucleotide consisting of the sequence GGGGGGGGGGGACGATCGTCGGGGGGGGGG (SEQ ID NO:6) is packaged, and wherein said unmethylated CpG containing oligonucleotide exclusively consists of phosphodiester connected deoxynucleotides. A preferred method of preparing QPGIO is described in Examples 1 and 8 of WO 2007/144150 or in Example 10 of WO 2007/144150.

"reliever medication" or "rescue medication": The terms "reliever medication" or "rescue medication" as interchangeably used herein, should refer to medications used on an as-needed basis that act quickly and immediately to reverse bronchoconstriction and relieve its symptoms. They include rapid-acting inhaled p₂-agonists, inhaled anticholinergics, short-acting theophylline, and short-acting oral ₂-agonists (SABA). The terms "reliever medication" or "rescue medication" as interchangeably used herein, typically and preferably, exclude controller treatment as defined herein. Thus, controller treatment and reliever medication, in the context of the present invention, are, typically and preferably, two different and distinguished treatments,

"somatotaxin"; Somatotaxins are well known immune modulating substances. Somatotaxins are synthetic ligands that bind to the 'CTLP site' on sstr2. They target the CTLP pathway and therefore mimic the anti-inflammatory effects of natural CTLPs implicated in the physiological resolution of inflammation. One example of somatotaxin is FX-125-L.

"stimulating production of IFN-alpha": IFN-alpha production by a cell can be determined by various methods generally known in the art, preferably by a method selected from (a) ELISA, most preferably by ELISA essentially as described in Example 14 of WO2007/068747; (b) flow cytometry analysis using fluorochrom-conjugated antibodies, preferably as described in Example 14 of WO2007/068747; and (c) cytopathicity inhibition bioassays. A typical cytopathicity inhibition bioassay is based on bovine MDBK cells infected with vesicular stomatitis virus, as previously described in Pestka, S. (1986) "Interferon Standards and General Abbreviations", in Methods in Enzymology, Academic Press, New York 119, 14- 23. Preferably, In the context of the present invention, an INA is regarded as being "capable of stimulating IFN-alpha production", when the production of IFN-alpha by a cell as detected by any one of the above described methods, preferably by ELISA, most preferably as described in Example 14 of WO2007/068747, is significantly increased upon exposure of said cell to said substance as compared to a control cell, wherein typically and preferably, said IFN-alpha production is increased by a factor of at least about 2, more preferably by a factor of about 3.

"toll-like receptor 7 ligand (TLR7-ligand)": As used herein, the term "toll-like receptor 7 ligand (TLR7-ligand)" refers to a ligand which is capable of activating TLR7 (Beutler, B., Curr Opin Hematol 9: 2-10 (2002); Schwarz, K., et al, Eur J Immunol 33: 1465-1470 (2003). TLR7 ligands are known to the skilled person in the art and include single stranded RNA, for example bacterial RNA, as well as synthetic substances such as imidazoquinolines (Diebold, S., et al, Science 303: 1529-1531 (2004); Heil, F. H., et al, Science 303: 1526-1528 (2004)). Preferred TLR7-ligands are imidazoquinolines such as imiquimod as well as AZD8848 or GSK 2245035.

"toll-like receptor 9 ligand (TLR9-ligand)": As used herein, the term "toll-like receptor 9 ligand (TLR9-ligand)" refers to a ligand which is capable of activating TLR9 (Beutler, B., Curr Opin Hematol 9: 2-10 (2002); Schwarz, K., et al, Eur J Immunol 33: 1465-1470 (2003), Abreu, M.T., and Arditi, M. J., Pediatrics 421-429 (2004)). TLR9 ligands are known to the skilled person in the art and include bacterial DNA, in particular DNA containing CpG motifs (Schnare et al. Curr Biol 10: 1139-42 (2000); Hemmi H., et al, Nature 408: 740-745 (2000); Krieg, A. M., Nat Rev Drug Discov 5(6):471-484 (2006)).

It is within a routine practice to test whether a ligand is a TLR7-ligand or a TLR9-ligand in accordance with the present invention, and whether a TLR ligand activates said TLR7 or TLR9. A typical and preferred example for such testing is as follows: 3xl0⁶ HEK293 cells are electroporated at 200 volt and 960 μΡ with 1 μg of TLR expression plasmid and 20 ng NF-kB luciferase reporter-plasmid. The overall amount of plasmid DNA is held constant at 15 μg per electroporation by addition of the appropriate empty expression vector. Cells are seeded at 105 cells per well and after overnight culture stimulated with the ligand to be tested for a further 7 to 10 hours. Typical examples of concentration ranges for known TLR ligands are 25 μg/ml RNA40-42 complexed to DOTAP (facilitating the internalization of RNA inside the cell), 1 μΜ CpG-ODN 2006, 10 μMR-848, 50 μg/ml poly(LC) or 1 μg/ml Pam3Cys (Heil, F. H., et al, Science 303: 1526-1528 (2004)). Stimulated cells are lysed using reporter lysis buffer (Promega, Mannheim, Germany) and lysate is assayed for luciferase activity using a luminometer, typically and preferably the Berthold luminometer (Wildbad, Germany), according to the manufacturer's instruction. It is within the knowledge of the skilled person in the art to accordingly adapt the aforementioned experiment for the testing of any TLR ligand. A ligand is, then, considered to activate a TLR in accordance with this invention, when the induced luciferase activity is higher, preferably statistically significantly higher, than a threshold value determined from the activitiy of the negative control (identical experiment and identical experimental conditions without the addition of the ligand to be tested). Preferably, a threshold value within this context is defined by the mean of the luciferase activities of the negative control in six independent experiments plus three times the standard deviation of the luciferase activities from the six experiments. A ligand is, then typically and preferably, considered to "statistically significantly" activate a TLR when the luciferase activity of the ligand is higher than the threshold value determined as indicated above. Preferably, a ligand is considered to "statistically significantly" activate a TLR when the luciferase activity of the ligand is at least two times higher, preferably three times higher, even more preferably five times higher than the threshold value determined as indicated above. The aforementioned typical and preferred example for such testing is in particular preferred for testing whether a ligand is a TLR7-ligand in accordance with the present invention.

A further typical and preferred example for testing whether a ligand is a TLR7-ligand or a TLR9-ligand in accordance with the present invention is described in Example 11 of WO2007/068747 where activation of bone marrow derived dendritic cells (BMDCs) is assessed. The aforementioned typical and preferred example, where induction of activation of BMDCs is assessed is the preferred manner to test whether a ligand is a TLR9-ligand in accordance with the present invention.

"uncontrolled": As used herein, the term "uncontrolled" refers to a patient having an ACQ-7 score of > 1.5. The 7-item Asthma Control Questionnaire ACQ-7 is a validated composite score based on the average of 7 items each scored 0-6, including patient's scores of 5 disease-related items (woken at night by asthma, awake in the morning with symptoms, limitation of daily activities, shortness of breath, wheezing) + 1 score for number of puffs of short acting p₂-agonists (SABA) + 1 score for FEVi predicted (Juniper, E. F., et al. Eur Respir J 14:32-38 (1999)).

"virus particle": The term "virus particle" as used herein refers to the morphological form of a virus. In some virus types it comprises a genome surrounded by a protein capsid; others have additional structures (e.g., envelopes, tails, etc.).

"virus-like particle (VLP)", as used herein, refers to a non-replicative or non-infectious, preferably a non-replicative and non-infectious virus particle, or refers to a non-replicative or non-infectious, preferably a non-replicative and non-infectious structure resembling a virus particle, preferably a capsid of a virus. The term "non-replicative", as used herein, refers to being incapable of replicating the genome comprised by the VLP. The term "non-infectious", as used herein, refers to being incapable of entering the host cell. Preferably a virus-like particle in accordance with the invention is non-replicative and/or non-infectious since it lacks all or part of the viral genome or genome function. In one embodiment, a virus-like particle is a virus particle, in which the viral genome has been physically or chemically inactivated, removed by disassembly and reassembly, or by assembly of purified proteins into a VLP. Typically and more preferably a virus-like particle lacks all or part of the replicative and infectious components of the viral genome. A virus-like particle in accordance with the invention may contain nucleic acid distinct from their genome. A typical and preferred embodiment of a virus-like particle in accordance with the present invention is a viral capsid such as the viral capsid of the corresponding virus, bacteriophage, preferably R A bacteriophage. The terms "viral capsid" or "capsid", refer to a macromolecular assembly composed of viral protein subunits. Typically, there are 60, 120, 180, 240, 300, 360 and more than 360 viral protein subunits. Typically and preferably, the interactions of these subunits lead to the formation of viral capsid or viral-capsid like structure with an inherent repetitive organization, wherein said structure is, typically, spherical or tubular. For example, the capsids of RNA bacteriophages or HBcAgs have a spherical form of icosahedral symmetry. The term "capsid-like structure" as used herein, refers to a macromolecular assembly composed of viral protein subunits resembling the capsid morphology in the above defined sense but deviating from the typical symmetrical assembly while maintaining a sufficient degree of order and repetitiveness. The invention encompasses VLPs, preferably non-natural VLPs, comprising a icosahedral symmetry. One common feature of virus particle and virus-like particle is its highly ordered and repetitive arrangement of its subunits.

"virus-like particle of a RNA bacteriophage": As used herein, the term "virus-like particle of a RNA bacteriophage" refers to a virus-like particle comprising, or preferably consisting essentially of or consisting of coat proteins, mutants or fragments thereof, of a RNA bacteriophage. Preferably, the term "virus-like particle of a RNA bacteriophage" refers to a virus-like particle comprising, or preferably consisting essentially of or consisting of coat proteins of a RNA bacteriophage. In another preferred embodiment, the term "virus-like particle of a RNA bacteriophage" refers to a virus-like particle consisting of coat proteins of a RNA bacteriophage. In another preferred embodiment, the term "virus-like particle of a RNA bacteriophage" refers to a virus-like particle comprising coat proteins of a RNA bacteriophage. In addition, virus-like particle of a RNA bacteriophage resembling the structure of a RNA bacteriophage, being non replicative and/or non-infectious, and lacking at least the gene or genes encoding for the replication machinery of the RNA bacteriophage, and typically also lacking the gene or genes encoding the protein or proteins responsible for viral attachment to or entry into the host. This definition should, however, also encompass virus-like particles of RNA bacteriophages, in which the aforementioned gene or genes are still present but inactive, and, therefore, also leading to non-replicative and/or non-infectious virus-like particles of a RNA bacteriophage. Preferred VLPs derived from RNA bacteriophages exhibit icosahedral symmetry and consist of 180 subunits. Within this present disclosure the term "subunit" and "monomer" are interexchangeably and equivalently used within this context. Preferred methods to render a virus-like particle of a RNA bacteriophage non replicative and/or non-infectious is by physical, chemical inactivation, such as UV irradiation, formaldehyde treatment, typically and preferably by genetic manipulation. Alternatively, individual proteins may be isolated from whole virions and assembled into VLPs in vitro.

"virosome": As used herein, the term virosome relates to a reconstituted virus envelope, preferably to a reconstituted influenza virus envelope, more preferably to a reconstituted envelope of influenza A virus, most preferably to a reconstituted envelope of influenza A/Singapore virus. Virosomes are known in the art and comprise a lipid membrane, wherein said lipid membrane typically and preferably comprises a unilamellar lipid bilayer. In a preferred meaning the term virosome relates to a reconstituted influenza virus envelope, preferably to a reconstituted influenza A virus envelope, most preferably to a reconstituted influenza A/Singapore virus envelope, wherein said reconstituted influenza virus envelope, preferably said reconstituted influenza A virus envelope, most preferably said reconstituted influenza A/Singapore virus envelope comprises lipid membrane, wherein said lipid membrane comprises influenza glycoproteins, wherein preferably said influenza glycoproteins are selected from hemagglutinin HA and neuraminidase NA.

"coat protein": As used herein, the term "coat protein(s)" refers to the protein(s) of a bacteriophage or a RNA bacteriophage capable of being incorporated within the capsid assembly of the bacteriophage or the RNA bacteriophage. However, when referring to the specific gene product of the coat protein gene of RNA bacteriophages the term "CP" is used. For example, the specific gene product of the coat protein gene of RNA bacteriophage QP is referred to as "QP CP", whereas the "coat proteins" of bacteriophage QP comprise the "QP CP" as well as the Al protein. The capsid of Bacteriophage QP is composed mainly of the QP CP, with a low content of the Al protein. Likewise, the VLP QP coat protein contains mainly QP CP, with a low content of Al protein.

"recombinant VLP": The term "recombinant VLP", as used herein, refers to a VLP that is obtained by a process which comprises at least one step of recombinant DNA technology. The term "VLP recombinantly produced", as used herein, refers to a VLP that is obtained by a process which comprises at least one step of recombinant DNA technology. Thus, the terms "recombinant VLP" and "VLP recombinantly produced" are interchangeably used herein and should have the identical meaning. "packaged": The term "packaged" as used herein refers to the state of an immune- modulating substance comprising, preferably consisting of an (i) INA, (ii) a TLR7-ligand, (iii) a TLR9-ligand or any mixture of at least one of (i), (ii) and/(iii), preferably to the state of an unmethylated CpG-containing oligonucleotide, in relation to the particle, preferably to the VLP. The term "packaged" as used herein refers to covalent binding, preferably by chemically coupling, and non-covalent binding. More preferably, the term "packaged" refers to non- covalent binding, preferably to ionic interactions, hydrophobic interactions, or hydrogen bonds. Very preferably, the term "packaged" as used herein refers to the enclosement, or partial enclosement, of said immune-modulating substance comprising, preferably consisting of (i) said INA, (ii) said TLR7-ligand, (iii) said TLR9-ligand or said any mixture of at least one of (i), (ii) and/(iii), within the particle. For example, the unmethylated CpG-containing oligonucleotide can be enclosed by the VLP without the existence of an actual binding, neither covalently nor non- covalently, or with a non-covalent binding.

Typically and preferably, a particle packaged with said immune-modulating substance comprising, preferably consisting of (i) said INA, (ii) said TLR7-ligand, (iii) said TLR9-ligand or said any mixture of at least one of (i), (ii) and/(iii), protects said immune -modulating substance comprising, preferably consisting of (i) said INA, (ii) said TLR7-ligand, (iii) said TLR9-ligand or said any mixture of at least one of (i), (ii) and/(iii), from degradation, preferably from DNAse or RNAse hydrolysis. Therefore, in the preferred meaning, the term "packaged" indicates that said immune-modulating substance comprising, preferably consisting of (i) said INA, (ii) said TLR7-ligand, (iii) said TLR9-ligand or said any mixture of at least one of (i), (ii) and/(iii), preferably said unmethylated CpG-containing oligonucleotide, in a packaged state is not accessible to DNAse or RNAse hydrolysis. More preferably, the term "packaged" indicates that said immune-modulating substance comprising, preferably consisting of (i) said INA, (ii) said TLR7-ligand, (iii) said TLR9-ligand or said any mixture of at least one of (i), (ii) and/(iii), preferably said unmethylated CpG-containing oligonucleotide, is not accessible to DNAse hydrolysis, wherein further preferably the DNAse is DNAsel or Benzonase. Still more preferably, the term "packaged" indicates that the unmethylated CpG-containing oligonucleotide is not accessible to Benzonase hydrolysis.

The accessibility of said immune -modulating substance comprising, preferably consisting of (i) said INA, (ii) said TLR7-ligand, (iii) said TLR9-ligand or said any mixture of at least one of (i), (ii) and/(iii), preferably of said unmethyated CpG-containing oligonucleotide, for DNAse (e.g. DNasel or Benzonase) is preferably assayed as described in Examples 11-17 of WO2003/024481 (see p. I l l therein). In a preferred meaning, a VLP is regarded as being packaged with an unmethylated CpG-containing oligonucleotide, when after treatment with Benzonase (190 U Benzonase / mg capsid protein in a buffer comprising 2 mM MgCk, pH 7.2, 20-25 °C, 18 h) at least 90 %, preferably at least 95 %, most preferably at least 98 % of said unmethylated CpG-containing oligonucleotide can be recovered from said VLP (e.g. in an ethidiumbromide stained gel). It is apparent for the artisan that such assays require appropriate controls and may need to be adapted to the specific combination of VLP and unmethylated CpG- containing oligonucleotide. In a more preferred meaning, a VLP of an RNA bacteriophage is regarded as being packaged with an unmethylated CpG-containing oligonucleotide, when after treatment with Benzonase (190 U Benzonase / mg capsid protein in a buffer comprising 2 mM MgCk, pH 7.2, 20-25 °C, 18 h) at least 90 %, preferably at least 95 %, most preferably at least 98 % of said unmethylated CpG-containing oligonucleotide can be recovered from said VLP of an RNA bacteriophage. In a very preferred meaning, a VLP of a RNA bacteriophage is regarded as being packaged with G10 (SEQ ID NO: 6) oligonucleotide, when after treatment with Benzonase (190 U Benzonase / mg capsid protein in a buffer comprising 2 mM MgCk , pH 7.2, 20-25 °C, 18 h) at least 90 %, preferably at least 95 %, most preferably at least 98 % of said unmethylated CpG-containing oligonucleotide can be recovered from said VLP of an RNA bacteriophage. In more specific meaning, a VLP of a RNA bacteriophage QP, AP205, GA or fr is regarded as being packaged with G10 (SEQ ID NO: 6) oligonucleotide, when after treatment with Benzonase (190 U Benzonase / mg capsid protein in a buffer comprising 2 mM MgCk , pH 7.2, 20-25 °C, 18 h) at least 90 %, preferably at least 95 %, most preferably at least 98 % of said unmethylated CpG-containing oligonucleotide can be recovered from said VLP of an RNA bacteriophage. In a very specific meaning, a VLP of a RNA bacteriophage QP is regarded as being packaged with G10 (SEQ ID NO:6) oligonucleotide, when after treatment with Benzonase (190 U Benzonase / mg capsid protein in a buffer comprising 2 mM MgCk , pH 7.2, 20-25 °C, 18 h) at least 90 %, preferably at least 95 %, most preferably at least 98 % of said unmethylated CpG-containing oligonucleotide can be recovered from said VLP of RNA bacteriophage QP.

Alternatively, the packaging state of said immune -modulating substance comprising, preferably consisting of (i) said INA, (ii) said TLR7-ligand, (iii) said TLR9-ligand or said any mixture of at least one of (i), (ii) and/(iii), in a particle, preferably of an unmethylated CpG- containing oligonucleotide, which do not constitute a substrate for DNAse or RNAse hydrolysis, can be assessed by size exclusion chromatography or SDS-PAGE and subsequent spectroscopic analysis as described in Example 4 of WO2007/068747. A further possibility to verify the packaging state of a particle packaged with said immune -modulating substance comprising, preferably consisting of (i) said INA, (ii) said TLR7-ligand, (iii) said TLR9-ligand or said any mixture of at least one of (i), (ii) and/(iii), preferably of an unmethylated CpG-containing oligonucleotide such as G10 (SEQ ID NO:6) is dialysis or tangential flow filtration of the particle, for example under conditions as described in Example 1 WO2007/068747, wherein non-packaged nucleic acids are removed while packaged nucleic aids remain associated with said particle.

In the very preferred meaning, and wherein the particle is a virus particle or a virus-like particle of a bacteriophage, preferably of an R A-bacteriophage, further preferably of an RNA bacteriophage Q , and most preferably of a virus-like particle of a RNA-bacteriophage Q , and wherein said immune -modulating substance is an unmethylated CpG-containing oligonucleotide, preferably a A-type CpG, further preferably the SEQ ID NO:6, the term "packaged" indicates that the particle packaged with said unmethylated CpG-containing oligonucleotide, preferably said A-type CpG, further preferably said SEQ ID NO: 6 elutes at the same retention time as the virus-like particle of said bacteriophage, preferably of said RNA-bacteriophage, further preferably of said RNA bacteriophage Q obtained by recombinant expression of the coat protein in E.coli, preferably wherein said retention time is determined by size exclusion chromatography, preferably as described in Example 4 of WO2007/068747, and comprises said said unmethylated CpG-containing oligonucleotide, preferably said A-type CpG, further preferably said SEQ ID NO: 6 as determined preferably as described in Example 4 of WO2007/068747.

In preferred embodiments, said immune-modulating substance comprising, preferably consisting of, said unmethylated CpG-containing oligonucleotide, is packaged inside the particle, preferably VLP capsids, most preferably in a non-covalent manner. Protocols for the preparation of VLPs packaged with unmethylated CpG-containing oligonucleotide are provided in the prior art, e.g. in WO2003/024481 (see Examples 2, 3, 7, 8, 10, 11, 12, 13, 14, 15, 16, and 17 therein, in particular Examples 14-17 therein) and WO2004/000351A1. The disclosure of both publications is incorporated to this application by reference. Further Protocols for the preparation of VLPs packaged with unmethylated CpG-containing oligonucleotide are provided Examples 1, 3 5 and 6 of WO2007/068747.

"polypeptide": As used herein the term "polypeptide" refers to a polymer composed of amino acid residues, generally natural amino acid residues, linked together through peptide bonds. Although a polypeptide may not necessarily be limited in size, the term polypeptide is often used in conjunction with peptide of a size of about ten to about 50 amino acids.

"protein": As used herein, the term protein refers to a polypeptide of a size of above 20, more preferably of above 50 amino acid residues. Proteins generally have a defined three dimensional structure although they do not necessarily need to, and are often referred to as folded, in contrast to peptides and polypeptides which often do not possess a defined three- dimensional structure, but rather can adopt a large number of different conformations, and are referred to as unfolded.

"sequence identity": The amino acid sequence identity of polypeptides can be determined conventionally using known computer programs such as the Bestfit program. When using Bestfit or any other sequence alignment program, preferably using Bestfit, to determine whether a particular sequence is, for instance, 95% identical to a reference amino acid sequence, the parameters are set such that the percentage of identity is calculated over the full length of the reference amino acid sequence and that gaps in homology of up to 5 % of the total number of amino acid residues in the reference sequence are allowed. This aforementioned method in determining the percentage of identity between polypeptides is applicable to all proteins, polypeptides or a fragment thereof disclosed in this invention.

"Sequence homology": The homology of nucleotide sequences is preferably determined by the program blastn which is an implementation of the BLAST algorithm, most preferably using the default settings of the software.

"fragment of a protein", in particular fragment of a recombinant protein or recombinant coat protein, as used herein, is defined as a polypeptide, which is of at least 70%, preferably at least 80%, more preferably at least 90%, even more preferably at least 95% the length of the wild-type recombinant protein, or coat protein, respectively and which preferably retains the capability of forming VLP. Preferably the fragment is obtained by at least one internal deletion, at least one truncation or at least one combination thereof. Preferably, the fragment is obtained by (i) at least one, preferably exactly one, internal deletion, (ii) at least one, preferably exactly one, truncation, or (Hi) at least one, preferably exactly one, combination thereof. Further preferably, the fragment is obtained by at most 5, 4, 3 or 2 internal deletions, at most 2 truncations or exactly one combination thereof. Further preferably, the fragment is obtained by at most 5, 4, 3 or 2 internal deletions, wherein still further preferably each of said deletions comprises 1 to 5, preferably 1 to 4, more preferably 1 to 3, still more preferably 1 to 2, and most preferably exactly 1 amino acid. The term "fragment of a recombinant protein" or "fragment of a coat protein" shall further encompass polypeptide, which has at least 80%>, preferably 90%>, even more preferably 95% amino acid sequence identity with the "fragment of a recombinant protein" or "fragment of a coat protein", respectively, as defined above and which is preferably capable of assembling into a virus-like particle.

The term "mutant coat protein" refers to a polypeptide having an amino acid sequence derived from the wild type recombinant protein, or coat protein, respectively, wherein the amino acid sequence is at least 80%, preferably at least 85%, 90%, 95%, 97%, or 99% identical to the wild type sequence and preferably retains the ability to assemble into a VLP. "allergen": The term "allergen" refers to a substance causing allergy. Preferred allergens are allergens disclosed in Shough, H. et al, REMINGTON'S PHARMACEUTICAL SCIENCES, 19th edition, (Chap. 82), Mack Publishing Company, Mack Publishing Group, Easton, Pennsylvania (1995), the entire contents of which is hereby incorporated by reference. The term "allergen", as used herein, also refers to "allergen extracts" and "allergenic epitopes." Very preferred allergens are selected from the group consisting of: pollens (e.g. grass, ragweed, birch and mountain cedar); house dust and dust mites; mammalian epidermal allergens and animal danders; mold and fungus; insect bodies and insect venom; feathers; food; and drugs (e.g. penicillin).

"aeroallergen": The term "aeroallergen" refers to any airborne substance, such as preferably to pollen or spores which triggers an allergic reaction Aeroallergens typically and preferably include plant pollen comprising from (i) trees such birch, alder, cedar, hazel, hornbeam, horse chestnut, willow, poplar, plane, linden/lime and olive (Olea). Grasses (Family Poaceae): especially ryegrass (Lolium sp.) and timothy (Phleum pratense). An estimated 90% of hay fever sufferers are allergic to grass pollen; (ii) weeds such as ragweed, plantain, nettles/parietaria, mugwort, Fat hen and sorrel/dock.

"effective amount": As used herein, the term "effective amount" refers to an amount necessary or sufficient to realize a desired biologic effect. An effective amount of the composition, or alternatively the pharmaceutical composition, would be the amount that achieves this selected result, and such an amount could be determined as a matter of routine by a person skilled in the art. Preferably, the term "effective amount", as used herein, refers to an amount necessary or sufficient to be effective to improve asthma control as defined herein. The effective amount can vary depending on the particular composition being administered, the size of the subject, and/or the severity of the disease or condition. One of ordinary skill in the art can empirically determine the effective amount of a particular composition of the present invention without necessitating undue experimentation.

"treatment": As used herein, the terms "treatment", "treat", "treated" or "treating" refer to prophylaxis and/or therapy. In one embodiment, the terms "treatment", "treat", "treated" or "treating" refer to a therapeutic treatment. In another embodiment, the terms "treatment", "treat", "treated" or "treating" refer to a prophylactic treatment.

"pharmaceutically acceptable carrier": The compositions of the invention can be combined, optionally, with a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" as used herein means one or more compatible solid or liquid fillers, diluents or encapsulating substances which are suitable for administration into a human. The term "carrier" denotes an organic or inorganic ingredient, natural or synthetic, with which the active ingredient is combined to facilitate the application.

"pharmaceutical composition": As used herein, the term "pharmaceutical composition" refers to a formulation which contains the composition of the invention and which is in a form that is capable of being administered to a human. Typically and preferably, the pharmaceutical composition comprises a conventional saline or buffered aqueous solution medium in which the composition of the present invention is suspended or dissolved. In this form, the composition of the present invention can be used conveniently to prevent, ameliorate, or otherwise treat a condition. Preferred pharmaceutical compositions induce, typically and preferably the formation of lFN-alpha.

"a/an": When the terms "a," or "an" are used in this disclosure, they mean "at least one" or "one or more," unless otherwise indicated.

As disclosed, in a first aspect, the invention provides a composition for use in a method for treating asthma of a patient in need thereof, wherein said method comprises (i) selecting a patient on controller treatment, wherein said patient is controlled by said patient's controller treatment, and wherein said patient having an eosinophilic cell count of greater than 0.1 cells/nl in peripheral blood; and (ii) administering an effective amount of said composition to said patient, wherein said composition is selected from (a) inhaled glucocorticosteriods (ICS); (b) long-acting p₂-agonists (LABA); (c) an immune -modulating substance; or (d) any mixtures of at least one of (a), (b) and/or (c); wherein said administration is effective to improve asthma control of said patient.

In a preferred embodiment, said patient has an eosinophilic cell count of equal to or greater than 0.15 cells/nl in peripheral blood, preferably said patient has an eosinophilic cell count of greater than 0.15 cells/nl in peripheral blood. In another preferred embodiment, said patient has an eosinophilic cell count of less than 0.45 cells/nl in peripheral blood. In another preferred embodiment, said patient has an eosinophilic cell count of equal to or less than 0.4 cells/nl in peripheral blood. In another preferred embodiment, said patient has an eosinophilic cell count of less than 0.25 cells/nl in peripheral blood. In another preferred embodiment, said patient has an eosinophilic cell count of less than 0.3 cells/nl in peripheral blood.

In another preferred embodiment, said patient has an eosinophilic cell count of greater than 0.1 cells/nl and equal to or less than 0.4 cells/nl in peripheral blood. In another preferred embodiment, said patient has an eosinophilic cell count of greater than 0.1 cells/nl and less than 0.3 cells/nl in peripheral blood. In another preferred embodiment, said patient has an eosinophilic cell count of equal to or greater than 0.15 cells/nl in peripheral blood and less than 0.45 cells/nl. In another preferred embodiment, said patient has an eosinophilic cell count of equal to or greater than 0.15 cells/nl and less than 0.25 cells/nl in peripheral blood.

In another preferred embodiment, patient's controller treatment is a treatment of GINA Step 2. In another preferred embodiment, said patient's controller treatment is a treatment of GINA Step 3. In a further more preferred embodiment said patient's controller treatment is treatment by way of inhaled glucocorticosteriods. In another preferred embodiment, said patient's controller treatment is treatment by way of low dose inhaled glucocorticosteriods. In another preferred embodiment, said patient's controller treatment is treatment by way of medium dose inhaled glucocorticosteriods. In another preferred embodiment, said patient's controller treatment is treatment by way of high dose inhaled glucocorticosteriods.

In another preferred embodiment, said patient's controller treatment is a treatment of GINA Step 4. In a further more preferred embodiment, said patient's controller treatment is treatment by way of inhaled corticosteroids combined with long-acting p₂-agonists. In another preferred embodiment, said patient's controller treatment is treatment by way of low dose inhaled glucocorticosteriods combined with long-acting ₂-agonists. In another preferred embodiment, said patient's controller treatment is treatment by way of medium dose inhaled glucocorticosteriods combined with long-acting ₂-agonists. In another preferred embodiment, said patient's controller treatment is treatment by way of high dose inhaled glucocorticosteriods combined with long-acting ₂-agonists.

In another preferred embodiment, said administering of said effective amount of said composition to said patient is repeated at least once, at least twice, at least three, four, five, six, or seven times, or at least ten times. In another preferred embodiment, said administering of said effective amount of said composition to said patient comprises repeated administrations of said effective amount of said composition to said patient, wherein said repeated administrations are effected in intervals of 1 week, 2 weeks, 3 weeks, monthly, bimonthly, quarterly or semiannually.

In a further again preferred embodiment, said patient's controller treatment is continued during said administering said effective amount of said composition to said patient. This, preferably, leads to a further improvement of the asthma control on top of patient's controller treatment.

In a further again preferred embodiment, said patient's controller treatment is reduced during said administering said effective amount of said composition to said patient. The administration of the composition of the invention, thus, preferably, leads to an improvement of the asthma control despite reduction of patient's controller treatment. In typical embodiments, reduction of patient's controller treatment is effected based on patient's asthma control. Preferably, said reduction of said patient's controller treatment is by 25%, 50% or 75%, more preferably by 50%. "Reduction of 25%, 50% or 75%" as used herein, means that the dosage of controller treatment is reduced by 25%, 50% or 75%. In a preferred embodiment, reduction of patient's controller treatment is 50%. In another preferred embodiment, said reduction of said patient's controller treatment is effected at least 12 weeks, preferably at least 8 weeks, more preferably at least 4 weeks after a first administration of said effective amount of said composition.

In a further again preferred embodiment, said patient's controller treatment is removed during said administering said effective amount of said composition to said patient. The administration of the composition of the invention, thus, preferably, allows despite removal of patient's controller treatment to improve asthma control. In another preferred embodiment, said removal of said patient's controller treatment is effected at least 12 weeks, preferably at least 8 weeks, more preferably at least 4 weeks after a first administration of said effective amount of said composition.

In another preferred embodiment, said administration is effective to improve asthma control of said patient as measured by either a reduction of ACQ-7 of at least of 0.3, preferably 0.5 points.

In another preferred embodiment, said composition comprises, preferably consists of, an inhaled glucocorticosteriod. In another preferred embodiment, said effective amount of said inhaled glucocorticosteriod is a low dose inhaled glucocorticosteriod. In another preferred embodiment, said effective amount of said inhaled glucocorticosteriod is a medium dose inhaled glucocorticosteriod. In another preferred embodiment, said effective amount of said inhaled glucocorticosteriod is a high dose inhaled glucocorticosteriod.

In another preferred embodiment, said composition comprises, preferably consists of, a combination of an inhaled glucocorticosteriod with a long-acting p₂-agonist. Preferably, said effective amount of said composition is a combination of a low dose inhaled glucocorticosteriod with a long-acting ₂-agonist, or said effective amount of said composition is a combination of a medium dose inhaled glucocorticosteriod with a long-acting ₂-agonist, or said effective amount of said composition is a combination of a high dose inhaled glucocorticosteriod with a long- acting ₂-agonist.

In another preferred embodiment, said composition comprises, preferably consists of, an immune-modulating substance, wherein preferably said immune-modulating substance comprises, preferably consists of, (i) an immune-modulating nucleic acid; (ii) a toll-like receptor 7 ligand (TLR7-ligand); (iii) a toll-like receptor 9 (TLR9-ligand); (iv) a chemoattractant receptor-homologous molecule expressed on Th2 cells antagonist (CRTh2 anatagonist); (v) a chemokine antagonist; (vi) a somatotaxin; or (vii) any mixture of at least one of (i), (ii), (iii), (iv), (v) and/or (vi).

In another further preferred embodiment, said composition comprises, preferably consists of, an immune-modulating substance, wherein preferably said immune-modulating substance comprises, preferably consists of, (i) an immune-modulating nucleic acid; (ii) TLR7-ligand; (iii) TLR9-ligand; or (iv) any mixture of at least one of (i), (ii) and/or (iii).

In another further preferred embodiment, said composition comprises, preferably consists of, an immune-modulating substance, wherein said immune-modulating substance comprises, preferably consists of, an immune-modulating nucleic acid. Preferably, said immune -modulating nucleic acid is capable of stimulating IFN-alpha production in a cell.

In another further preferred embodiment, said immune-modulating nucleic acid is selected from (a) a desoxyribonucleic acid; (b) a ribonucleic acid, (c) a chimeric nucleic acid; or (d) any mixture of at least one nucleic acid of (a), (b) and/or (c).

In another very preferred embodiment, said immune -modulating nucleic acid comprises, preferably consists of, an unmethylated CpG containing oligonucleotide. In another embodiment, said ribonucleic acid comprises, preferably consists of, poly-(LC) or a derivative thereof. . In another embodiment, said deoxyribonucleic acid comprises, preferably consists of, an oligonucleotide free of an unmethylated CpG motif.

In another further preferred embodiment, said composition comprises, preferably consists of, an immune-modulating substance, wherein said immune-modulating substance comprises, preferably consists of, a TLR7-ligand. In another further preferred embodiment, said composition comprises, preferably consists of, an immune-modulating substance, wherein said immune-modulating substance comprises, preferably consists of, a TLR9-ligand.

In another very preferred embodiment, said TLR9-ligand comprises, preferably consists of, an unmethylated CpG containing oligonucleotide.

In another very preferred embodiment, said composition comprises, preferably consists of, an immune-modulating substance, wherein said immune -modulating substance comprises, preferably consists of an unmethylated CpG containing oligonucleotide.

Preferably, said unmethylated CpG containing oligonucleotide is selected from of (a) A- type CpGs; (b) B-type CpGs; or (c) C-type CpGs. Preferably, said unmethylated CpG-containing oligonucleotide comprises a palindromic sequence, wherein preferably the CpG motif of said unmethylated CpG-containing oligonucleotide is part of a palindromic sequence, and wherein further preferably said palindromic sequence is GACGATCGTC (SEQ ID NO: l).

In a very preferred embodiment of the present invention, said unmethylated CpG- containing oligonucleotide comprises, or alternatively consists essentially of, or alternatively consists of the sequence GGGGGGGGGGGACGATCGTCGGGGGGGGGG (SEQ ID NO:6), wherein further preferably said unmethylated CpG-containing oligonucleotide consists exclusively of phosphodiester bound nucleotides.

Typically and preferably, said immune-modulating nucleic acid, preferably said unmethylated CpG-containing oligonucleotide, is not accessible to DNAse hydrolysis or is not accessible to Benzonase hydrolysis.

In a very preferred embodiment of the present invention, said immune-modulating nucleic acid is an unmethylated CpG containing oligonucleotide consisting of the sequence GGGGGGGGGGGACGATCGTCGGGGGGGGGG (SEQ ID NO: 6), and wherein said unmethylated CpG-containing oligonucleotide consists exclusively of phosphodiester bound nucleotides.

In a further preferred embodiment of the present invention, said composition comprises, preferably consists of, an immune-modulating substance, wherein said immune-modulating substance comprises, preferably consists of (i) an immune -modulating nucleic acid; (ii) a TLR7- ligand; (iii) a TLR9-ligand; or (iv) any mixture of at least one of (i), (ii) and/or (iii), wherein said immune -modulating substance is packaged in a particle, wherein said particle is, preferably, a virus-like particle or a virosome.

In another embodiment, said particle is a virosome, wherein preferably said visosome is a reconstituted virus envelope of an influenza virus, wherein further preferably said influenza virus is an influenza A virus, wherein still further preferably said influenza A virus is influenza A/Singapore virus. Virosomes comprising cationic (positively charged) lipids are especially suited. In a further embodiment, said particle is a virosome, wherein said virosome comprises a lipid membrane, wherein said lipid membrane comprises or preferably essentially consists of cationic lipids. In a preferred embodiment, said particle is a virosome and said immune - modulating substance is a unmethylated CpG-containing oligonucleotide, wherein preferably said unmethylated CpG containing oligonucleotide is G10 (SEQ ID NO: 6), and wherein further preferably said virosome comprises a lipid membrane, wherein said lipid membrane comprises or preferably essentially consists of cationic lipids.

In a preferred embodiment said particle is a virus-like particle (VLP). Any virus known in the art may be selected as a VLP of the invention. Most commonly known viruses have been sequenced and are readily available to the public. The taxonomy of viruses is well known to the artisan and summarized, for example, in H.V. Van Regenmortel et al. (eds.), Virus Taxonomy: 7^th Report of the International Committee on Taxonomy of Viruses (2000) (Academic Press/elsevier, Burlington Mass, USA), on the Virus Taxonomy web-page of the University of Leicester (UK) (http://www-micro.msb. le.ac.uk/3035/Vimsgroups.html) and by the Taxonomy Browser of the National Center for Biotechnology Information (NCBI, Washington D.C., USA) (http://www.ncbi.nlm.nih.gov/ICTVdb/). The genes encoding viral coat proteins can be identified by a skilled artisan and their nucleotide and amino acid sequences may, for example, be obtained from Genbank (http://www.ncbi.nlm.nih.gov/). Viruses which are particularly useful in the context of the invention are generally disclosed in "Artificial DNA - Methods and Applications", Yury Khudyakov and Howard Fields, eds., CCR Press, 2003.

VLPs can be produced and purified from virus-infected cell cultures. For the purpose of the present invention, said VLPs are non-replicative or non-infectious, preferably non-replicative and non-infectious. UV irradiation, chemical treatment, such as with formaldehyde, β-propione or chloroform, are the general methods known to skilled person to inactivate a virus. Alternatively, said non-replicative and non-infectious VLP can be produced by purification and reassembly of core proteins of said virus.

In another embodiment said particle is a VLP of a virus, wherein said virus is a DNA virus. In another embodiment said DNA virus is a single stranded DNA virus, wherein said single stranded DNA virus is preferably from: (a) Parvovirus, preferably parvovirus B19, porcine parvovirus (PPV) or canine parvovirus (CPV), (b) Erythrovirus, (c) Dependovirus, (d) recombinant of CPV with feline panleucopenia virus (FPV) (Saliki, T. J. et al., J Gen Virol 73 :369ff (1992)), (e) adeno-associated virus type 2 (AAV-2), (f) mink enteritis parvovirus (MEV), (g) muscovy duck parvovirus (DPV), (h) minute virus of mice (MVM), (i) aleutian mink disease parvovirus (ADV), or (j) Galleria mellonella densovirus (GMDNV).

In another embodiment said DNA virus is a double stranded DNA virus, including double stranded DNA reverse transcribing viruses, wherein said double stranded DNA virus is preferably selected from: (a) nucleopolyhedrovirus, preferably Autograpa californica nucleopolyhedrovirus (AcMNPV) or a chimera of AcMNPV polyhedrin and Trichoplusioa ni granulosis virus (TnGV) (Eason J.E. et al. (1998), J Virol 72:6237ff), (b) papillomavirus, preferably selected from (i) human papilloma virus (HPV, most preferably HPV6, HPV1 1 , HPV16, HPV18, or HPV33), (ii) bovine papillomavirus (BPV, preferably BPV1), and (Hi) cottontail rabbit papillomavirus (CRPV), (c) polyomavirus, preferably selected from (i) murine polyomavirus (preferably Py or SV40), (ii) budgerigar fledgling virus, (Hi) human polyomavirus JC, (iv) hamster polyomavirus (HaPV), (v) monkey B-lympotropic papovirus (LPV), (vi) avian polyomavirus (APV) and (vii) recombinant human and non-human polyomaviruses (Sasnauskas K. et al (1999) Biol. Chem. 380, 381), (d) spleen necrosis virus (SNV, Jiang A. (1999) Hum. Gene Therapy 10(16):2627-2636), or, very preferably, (e) Hepatitis B virus.

In another embodiment, the VLP is a VLP of Hepatitis B virus. The preparation of Hepatitis B virus-like particles has been disclosed, inter alia, in WO00/32227, WOO 1/85208, WO01/056905 and WO2004/000351. All four documents are explicitly incorporated herein by way of reference.

In a further preferred embodiment said virus is a RNA virus, wherein said R A virus is a single stranded RNA virus or a double stranded RNA virus. In a further preferred embodiment said RNA virus is a single stranded RNA virus, wherein preferably said single stranded RNA virus is a single stranded positive sense RNA virus, wherein preferably said single stranded positive sense RNA virus is selected from: (a) bromoviridae, preferably selected from (i) alfamovirus (e.g. alfalfa mosaic virus (A1MV)), and (ii) ilarvirus (e.g. prunus necrotic ringspot ilarvirus (PNRSV, Pallas V. (1998) Arch. Virol. 144:797-803); prune dwarf virus (PDV, Abou- Jawdah Y. et al. (2004) J. Virological Methods 121 :31-38)), (Hi) bromovirus (e.g. cowpea chlorotic mottle virus (CCMV) or brome mosaic virus (BMV)), (iv) cucumovirus (e.g. cucumber mosaic virus, Natilla A. et al. Arch Virol 2004 149(1): 137-154), (b) tombusviridae, preferably (i) tombusvirus, preferably tomato bushy stunt virus (TBSV, Joelson T. et al. (1997) J. Gen. Virol. 78: 1213-1217), (ii) carmovirus, turnic crinkle virus (TCV, Qu F. and Morris T.J. (1997) J. Virol. 71(2): 1428-1435), (c) potyvirus, preferably Johnsongrass mosaic virus (JGMV) and plum pox potyvirus (PPV, Fernandez-Fernandes M.R. et al. (2002) J. Virol. 76(24): 12646-12653), (d) tobacco mosaic virus (TMV), (e) comovirus, preferably cowpea mosaic virus (CPMV), (f) potato virus X (PVX, Marusic C. et al. (2001) J. Virol. 75(18):8434-8439, (g) calicivirus, preferably selected from (i) norwalk virus (NV), (ii) norwalk-like calcivirus, (Hi) human calcivirus, (iv) Lorsdale calcivirus, (v) rabbit hemorrhagic disease virus (RHDV), (vi) European brown hare syndrom virus (EBHSV), (vii) Toronto virus, (viii) Hawaii virus, (ix) Sapporo-like virus, and (x) Grimsby feline calcivirus, (h) RNA bacteriophage, (i) luteovirus, preferably potato leaf roll virus (PLRV), (j) flock house virus, (k) retroid viruses, preferably selected from (i) oncoretrovirus, (ii) lentivirus, and (Hi) yeast retrotransposon Tyl , (1) tick-borne encephalitis virus (TBEV, Leibl H. (1998) Vaccine 16(4):340-345) or (m) togaviridae, preferably alphavirus, most preferably Sindbis virus.

In a further preferred embodiment said RNA virus is a single stranded positive sense RNA virus selected from: (a) bromoviridae, preferably selected from (i) alfamovirus (e.g. alfalfa mosaic virus (A1MV)), and (ii) ilarvirus (e.g. prunus necrotic ringspot ilarvirus (PNRSV, Pallas V. (1998) Arch. Virol. 144:797-803); prune dwarf virus (PDV, Abou- Jawdah Y. et al. (2004) J. Virological Methods 121 :31-38)), (Hi) bromovirus (e.g. cowpea chlorotic mottle virus (CCMV) or brome mosaic virus (BMV)), (iv) cucumovirus (e.g. cucumber mosaic virus, Natilla A. et al. Arch Virol 2004 149(1): 137-154), (b) tombusviridae, preferably (i) tombusvirus, preferably tomato bushy stunt virus (TBSV, Joelson T. et al. (1997) J. Gen. Virol. 78: 1213-1217), (ii) carmovirus, turnic crinkle virus (TCV, Qu F. and Morris T.J. (1997) J. Virol. 71(2): 1428-1435), (c) potyvirus, preferably Johnsongrass mosaic virus (JGMV) and plum pox potyvirus (PPV, Fernandez-Fernandes M.R. et al. (2002) J. Virol. 76(24): 12646-12653), (d) tobacco mosaic virus (TMV), (e) comovirus, preferably cowpea mosaic virus (CPMV), (f) potato virus X (PVX, Marusic C. et al. (2001) J. Virol. 75(18):8434-8439, (g) calicivirus, preferably selected from (i) norwalk virus (NV), (ii) norwalk-like calcivirus, (Hi) human calcivirus, (iv) Lorsdale calcivirus, (v) rabbit hemorrhagic disease virus (RHDV), (vi) European brown hare syndrom virus (EBHSV), (vii) Toronto virus, (viii) Hawaii virus, (ix) Sapporo-like virus, and (x) Grimsby feline calcivirus, (h) RNA bacteriophage, (i) luteovirus, preferably potato leaf roll virus (PLRV), (j) flock house virus, (k) retroid viruses, preferably selected from (i) oncoretrovirus, (ii) lentivirus, and (Hi) yeast retrotransposon Tyl , (1) tick-borne encephalitis virus (TBEV, Leibl H. (1998) Vaccine 16(4):340-345), (m) togaviridae, preferably alphavirus, most preferably Sindbis virus, or (n) Nodaviridae, preferably Alphanodavirus, most preferably Pariacoto virus (Johnson K.N. et al. (2004) Journal of Virology 78: 1 1371-1 1378).

In a further preferred embodiment said RNA virus is a double stranded RNA virus, wherein preferably said double stranded RNA virus is selected from: (a) birnavirus, (b) cypovirus, preferably Bombyx mori cytoplasmic polyhedrovirus (BmCPV), (c) orbivirus, preferably bluetoung virus (BTV) or African horse sickness virus (AHSV), (d) rotavirus and, very preferably, (e) double stranded RNA bacteriophages, preferably selected from (i) bacteriophage 8, (ii) bacteriophage phi6, (Hi) bacteriophage phil2, or (iv) bacteriophage phil2.

In a further preferred embodiment said virus particle or VLP is a virus particle or VLP of a virus, wherein said virus is a bacteriophage, wherein said bacteriophage is a DNA bacteriophage or an RNA bacteriophage.

In a preferred embodiment said bacteriophage is a DNA bacteriophage, wherein said DNA bacteriophage is a single stranded DNA bacteriophage or a double stranded bacteriophage. In a preferred embodiment, said DNA bacteriophage is a single stranded DNA bacteriophage, wherein said single stranded DNA bacteriophage is preferably selected from (a) Micro viridae, preferably Phi X 174 and (b) Inoviridae, preferably fd and M13. In a further preferred embodiment, said DNA bacteriophage is a double stranded DNA bacteriophage, wherein said double stranded DNA bacteriophage is preferably selected from: (a) Myoviridae, preferably T2, T4 or T6, (b) Siphoviridae, preferably bacteriophage Lambda, Tl , T5 or HK97, (c) Podoviridae, preferably T2, T7 or P22, (d) Tectiviridae, preferably PRD1 , (e) Corticoviridae, preferably PM2, (f) Plasmaviridae, preferably mycoplasma phages, (g) Lipothrixviridae, preferably Thermoproteus bacteriophage TTV1 or (h) Fuselloviridae, preferably sulfolobus bacteriophage 1. In a more preferred embodiment said bacteriophage is an R A bacteriophage, wherein said RNA bacteriophage is a single stranded or a double stranded RNA bacteriophage. In one embodiment said RNA bacteriophage is a single stranded RNA bacteriophage, wherein preferably said single stranded RNA bacteriophage is an enterobacteriophage, wherein preferably said enterobacteriophage is a representative of the Leviviridae, wherein preferably said representative of the Leviviridae is selected from: (a) taxonomically not assigned family member Acinetobacter phage 205 (AP205), (b) levivirus, or, preferably (c) allolevivirus.

In a preferred embodiment said representative of the Leviviridae is a levivirus, wherein preferably said levivirus is selected from: (a) bacteriophage BZ13, (b) bacteriophage GA, (c) bacteriophage JP34, (d) bacteriophage KU1, (d) bacteriophage TH1, (e) bacteriophage MS2, (f) bacteriophage f2, (g) bacteriophage fr, (h) bacteriophage JP501, (i) bacteriophage M12, (j) bacteriophage R17, or (k) bacteriophage PP7.

In a more preferred embodiment said representative of the Leviviridae is an allolevivirus, wherein preferably said allolevivirus is selected from: (a) bacteriophage FI, (b) bacteriophage ID2, (c) bacteriophage NL95, (d) bacteriophage SP, (d) bacteriophage TW28, (e) bacteriophage QP, (f) bacteriophage Mi l, (g) bacteriophage MX1, (h) bacteriophage ST, (i) bacteriophage TW18, or (j) bacteriophage VK.

In a very preferred embodiment of the present invention, said particle is a virus-like particle of RNA bacteriophage QP.

In a further preferred embodiment said RNA bacteriophage is selected from: (a) bacteriophage BZ13, (b) bacteriophage GA, (c) bacteriophage JP34, (d) bacteriophage KU1, (d) bacteriophage TH1, (e) bacteriophage MS2, (f) bacteriophage f2, (g) bacteriophage fr, (h) bacteriophage JP501, (i) bacteriophage M12, (j) bacteriophage R17, (k) bacteriophage PP7, (1) bacteriophage FI, (m) bacteriophage ID2, (n) bacteriophage NL95, (o) bacteriophage SP, (p) bacteriophage TW28, (q) bacteriophage QP, (r) bacteriophage Mi l, (s) bacteriophage MX1, (t) bacteriophage ST, (u) bacteriophage TW18, or (v) bacteriophage VK. In a further preferred embodiment said RNA bacteriophage is selected from: (a) bacteriophage QP, (b) bacteriophage R17, (c) bacteriophage fr, (d) bacteriophage GA, (e) bacteriophage SP, (f) bacteriophage MS2, (g) bacteriophage Mi l, (h) bacteriophage MX1, (i) bacteriophage NL95, (k) bacteriophage f2, (1) bacteriophage PP7, or (m) bacteriophage AP205.

In a further preferred embodiment said RNA bacteriophage is a double stranded RNA bacteriophage, wherein preferably said double stranded RNA bacteriophage is a representative of the Cystoviridae, more preferably said representative of the Cystoviridae is a Cystovirus, most preferably said Cystovirus is pseudomonas bacteriophage Phi 6. In a preferred embodiment said particle is a virus particle of a bacteriophage, and wherein preferably said bacteriophage is a RNA bacteriophage, wherein further preferably said RNA bacteriophage is a single stranded positive sense RNA bacteriophage, and wherein still further preferably said single stranded positive sense RNA bacteriophage is a single stranded positive sense RNA bacteriophage selected from: (a) bacteriophage QP, (b) bacteriophage fr, (c) bacteriophage GA, and (d) bacteriophage AP205, most preferably said single stranded positive sense RNA bacteriophage is Qp.

In a very preferred embodiment, said particle is a VLP, preferably a VLP of an RNA virus, more preferably a VLP of a single stranded positive sense RNA virus, most preferably a VLP of an RNA bacteriophage. In an again more preferred embodiment said particle is a VLP of an RNA bacteriophage QP.

In a further preferred embodiment said particle is a VLP of a bacteriophage, more preferably a VLP of a enterobacteriophage, still more preferably a VLP of a representative of the Leviviridae, most preferably a VLP of a levivirus or an allolevivirus. In a very preferred embodiment said VLP is a VLP of an allolevivirus.

In a further embodiment said particle is a VLP of a icosahedral virus, wherein said icosahedral virus is preferably a plant-infectious icosahedral virus. VLPs of plant-infectious icosahedral viruses are for example disclosed in WO2005/067478A2 which is incorporated herein by reference. In a preferred embodiment said icosahedral virus is selected from a representative of any one taxon selected from (a) Papillomaviridae, (b) Totiviridae, (c) Dcistroviridae, (d) Hepadnaviridae, (e) Togaviridae, (f) Polyomaviridae, (g) Nodaviridae, (h) Tectiviridae, (i) Leviviridae, (j) Microviridae, (k) Sipoviridae, (1) Picornoviridae, (m) Parvoviridae, (n) Calciviridae, (o) Tetraviridae, or (p) Satellite viruses. In another embodiment, said icosahedral virus is a plant-infectious icosahedral virus, wherein said plant-infectious icosahedral virus is a representative of any one taxon selected from (a) Bunyaviridae, (b) Reoviridae, (c) Rhabdoviridae, (d) Luteoviridae, (e) Nanoviridae, (f) Partitiviridae, (g) Sequiviridae, (h) Tymoviridae, (i) Ourmiavirus, (j) Tobacco Necrosis Virus Satellite, (k) Caulimoviridae, (1) Geminiviridae, (m) Comoviridae, (n) Sobemovirus, (o) Tombusviridae, or (p) Bromoviridae. In another embodiment, said plant-infectious icosahedral virus is a representative of any one taxon selected from (a) Luteoviridae, (b) Nanoviridae, (c) Partitiviridae, (d) Sequiviridae, (e) Tymoviridae, (f) Ourmiavirus, (g) Tobacco Necrosis Virus Satellite, (h) Caulimoviridae, (i) Geminiviridae, (j) Comoviridae, (k) Sobemovirus, (1) Tombusviridae, or (m) Bromoviridae. In another embodiment, said plant-infectious icosahedral virus is a representative of any one taxon selected from (a) Caulimoviridae, (b) Geminiviridae, (c) Comoviridae, (d) Sobemovirus, (e) Tombusviridae, or (e) Bromoviridae. In another embodiment, said plant-infectious icosahedral virus is a representative of any one taxon selected from (a) Comoviridae, (b) Sobemovirus, (c) Tombusviridae, or (d) Bromoviridae. In another embodiment, said plant-infectious icosahedral virus is a representative of any one taxon selected from Comoviridae or Bromoviridae. In another embodiment said plant-infectious icosahedral virus is a Cowpea Mosaic Virus or a Cowpea Chlorotic Mottle Virus. In another embodiment said plant-infectious icosahedral virus is a representative of the Bromoviridae, preferably Bromovirus, Cucumovirus, Ilarvirus or Alfamovirus. In another embodiment said plant- infectious icosahedral virus is selected from: brome mosaic virus, cowpea chlorotic mottle virus, cucumber mosaic virus, Tobacco streak virus or alfalfa mosaic virus (AMV, including AMV1 and AMV2).

In a preferred embodiment, the VLP is a recombinant VLP. The preparation of VLPs by recombinantly expressing the coat protein in a host is within the common knowledge of a skilled artisan. Illustrative DNA or RNA viruses, the coat or capsid protein of which can be used for the preparation of VLPs have been disclosed in WO 2004/009124 on page 25, line 10-21, on page 26, line 11-28, and on page 28, line 4 to page 31, line 4. These disclosures are incorporated herein by way of reference.

In one preferred embodiment, said VLP comprises, or alternatively consists of, recombinant proteins, mutants or fragments thereof, of a virus, wherein preferably said virus is selected from any virus listed above. In a very preferred embodiment said VLP comprises, or alternatively consists of, recombinant proteins, mutants or fragments thereof, of a virus, wherein said virus is selected from (a) RNA bacteriophages, (b) bacteriophages, (c) Hepatitis B virus, preferably its capsid protein (Ulrich, et al, Virus Res. 50: 141-182 (1998)) or its surface protein (WO 92/11291), (d) measles virus (Warnes, et al, Gene 160:173-178 (1995)), (e) Sindbis virus; (f) rotavirus (US 5,071,651 and US 5,374,426), (g) foot-and-mouth-disease virus (Twomey, et al, Vaccine 13: 1603 1610, (1995)), (h) Norwalk virus (Jiang, X., et al, Science 250: 1580 1583 (1990); Matsui, S.M., et al, J. Clin. Invest. 87: 1456 1461 (1991)), (i) Alphavirus, (j) retrovirus, preferably its GAG protein (WO 96/30523), (k) retrotransposon Ty, preferably the protein pi; (1) human Papilloma virus (WO 98/15631), (m) Polyoma virus, (n) Tobacco mosaic virus, or (o) Flock House Virus. In another embodiment said VLP comprises, or alternatively consists of, recombinant proteins, mutants or fragments thereof, of a virus, wherein said virus is selected form (a) Hepatitis B virus, or (b) Polyoma virus.

In another embodiment, said VLP comprises, or alternatively consists of, recombinant proteins, mutants or fragments thereof, of a virus, wherein said virus is a plant-infectious icosahedral virus, wherein preferably said plant-infectious icosahedral virus is selected from (a) Comoviridae, (b) Sobemovirus, (c) Tombusviridae, or (d) Bromoviridae. In another embodiment, the VLP is a VLP of Hepatitis B virus. The preparation of Hepatitis B virus-like particles has been disclosed, inter alia, in WO00/32227, WOO 1/85208, WOO 1/056905 and WO2004/000351. All four documents are explicitly incorporated herein by way of reference.

In one preferred embodiment of the invention, the virus-like particle comprises, consists essentially of, or alternatively consists of, recombinant coat proteins, mutants or fragments thereof, of a RNA bacteriophage. Preferably, the RNA bacteriophage is selected from (a) bacteriophage BZ13, (b) bacteriophage GA, (c) bacteriophage JP34, (d) bacteriophage KU1, (d) bacteriophage TH1, (e) bacteriophage MS2, (f) bacteriophage f2, (g) bacteriophage fr, (h) bacteriophage JP501, (i) bacteriophage M12, (j) bacteriophage R17, (k) bacteriophage PP7, (1) bacteriophage FI, (m) bacteriophage ID2, (n) bacteriophage NL95, (o) bacteriophage SP, (p) bacteriophage TW28, (q) bacteriophage QP, (r) bacteriophage Mi l, (s) bacteriophage MX1, (t) bacteriophage ST, (u) bacteriophage TW18, or (v) bacteriophage VK. Further preferably, the RNA bacteriophage is selected from (a) bacteriophage QP, (b) bacteriophage R17, (c) bacteriophage fr, (d) bacteriophage GA, (e) bacteriophage SP, (f) bacteriophage MS2, (g) bacteriophage Mi l, (h) bacteriophage MX1, (i) bacteriophage NL95, (k) bacteriophage f2, (1) bacteriophage PP7, or (m) bacteriophage AP205.

In one preferred embodiment of the invention, the virus-like particle comprises at least one coat protein of an RNA bacteriophage, wherein the RNA bacteriophage is selected from (a) bacteriophage QP, (b) bacteriophage R17, (c) bacteriophage fr, (d) bacteriophage GA, (e) bacteriophage SP, (f) bacteriophage MS2, (g) bacteriophage Mi l, (h) bacteriophage MX1, (i) bacteriophage NL95, (k) bacteriophage f2, (1) bacteriophage PP7, or (m) bacteriophage AP205.

In a further very preferred embodiment of the present invention, the virus-like particle comprises coat proteins having an amino acid sequence selected from (a) SEQ ID NO:7; and (b) a mixture of SEQ ID NO:7 and SEQ ID NO:8 (QP Al protein).

In a further very preferred embodiment of the present invention, the virus-like particle consists of coat proteins having an amino acid sequence of SEQ ID NO:7, or consists of a mixture of coat proteins having amino acid sequences of SEQ ID NO: 8, or mutants thereof, and of SEQ ID NO:7.

In a still further very preferred embodiment of the present invention, the virus-like particle consists of coat proteins consisting of an amino acid sequence of SEQ ID NO:7, or consists of a mixture of coat proteins consisting of amino acid sequences of SEQ ID NO: 8 and of SEQ ID NO:7.

In one embodiment, the particle is a VLP of bacteriophage fr or GA. Fr coat protein in the form of recombinant VLP may be obtained as described by Pushko P et al. ((1993) Prot Engin 6:883-891), while GA VLP may be obtained by cloning GA coat protein cDNA isolated by reverse transcription from GA phage into pQbl85, which is described for example in WO2004/007538. Disassembly of Fr and GA VLPs can be readily done by incubating the VLPs in 7 M urea, optionally supplemented with acetic acid at a concentration of 0.1 M. The nucleic acid is further purified from the coat protein by ion exchange chromatography, either at a pH where a significant amount of the coat protein flows through while the nucleic acid is retained, or at a pH where the coat protein is also adsorbed on the column and subsequently eluted with a salt gradient. Reassembly of fr and GA coat protein with INA is effected essentially as described in WO2003/024481 by slow dialysis, wherein said INA preferably is an unmethylated CpG- containing oligonucleotide, more preferably G10 (SEQ ID NO:6), and even more preferably an aggregated G10 (SEQ ID NO:6) as described in Example 2 of WO2007/068747, again more preferably as described in Example 1 of WO 2007/144150. Further protocols for the disassembly and reassembly of Fr and GA VLPs are disclosed in Examples 5 and 6 of WO2007/068747.

In preferred embodiments of the present invention, the virus-like particle of the invention comprises, or alternatively consists essentially of, or alternatively consists of recombinant coat proteins, mutants or fragments thereof, of the RNA-bacteriophage QP, fr, AP205 or GA.

In a very preferred embodiment, the VLP of the invention is a VLP of RNA bacteriophage QP. The capsid or virus-like particle of QP showed an icosahedral phage-like capsid structure with a diameter of 25 nm and T=3 quasi symmetry. The capsid contains 180 copies of the coat protein, which are linked in covalent pentamers and hexamers by disulfide bridges (Golmohammadi, R. et al., Structure 4:543-5554 (1996)), leading to a remarkable stability of the QP capsid. In an again more preferred, embodiment, said particle is a virus-like particle of an RNA-phage QP consisting of coat proteins with the amino acid sequence of SEQ ID NO:7.

Further preferred VLPs of RNA bacteriophages in accordance with this invention, in particular of QP and fr, are disclosed in WO 02/056905, the disclosure of which is herewith incorporated by reference in its entirety. In particular Example 18 of WO 02/056905 gave detailed description of preparation of VLP particles from QP.

In another preferred embodiment, said particle is a VLP of RNA bacteriophage AP205. Assembly-competent mutant forms of AP205 VLPs, including AP205 coat protein with the substitution of proline at amino acid 5 to threonine, may also be used in the practice of the invention and leads to other preferred embodiments of the invention. WO 2004/007538 describes, in particular in Example 1 and Example 2, how to obtain VLP comprising AP205 coat proteins, and hereby in particular the expression and the purification thereto. WO 2004/007538 is incorporated herein by way of reference. In a further preferred embodiment said virus particle or VLP is a virus particle or VLP of RNA bacteriophage AP205, wherein said INA preferably is an unmethylated CpG-containing oligonucleotide, more preferably G10 (SEQ ID NO:6), and even more preferably aggregated G10 (SEQ ID NO: 6) as described in Example 1 of WO 2007/144150. The disassembly and reassembly of AP205 is demonstrated in Example 5 of WO2007/068747.

QP mutants, of which exposed lysine residues are replaced by arginines can be used for the present invention. The construction, expression and purification of such preferred QP mutant coat proteins, mutant QP coat protein VLPs are described in WO 02/056905, in particular as described in Example 18 of WO 02/056905.

In a very preferred embodiment, said particle is a VLP of RNA bacteriophage QP, wherein said INA preferably is an unmethylated CpG-containing oligonucleotide, more preferably G10 (SEQ ID NO:6), and even more preferably aggregated G10 (SEQ ID NO:27) having a retention time relative to QP capsid standard under HPLC conditions, as set forth in Example 1 of WO2007/144150, of 50 to 110 %, most preferably of 80 to 110 %. The disassembly and reassembly of QP VLPs is demonstrated in Example 8 or Example 10 of WO2007/144150.

Further RNA bacteriophage coat proteins have also been shown to self-assemble upon expression in a bacterial host (Kastelein, RA. et al., Gene 23:245-254 (1983), Kozlovskaya, TM. et al, Dokl. Akad. Nauk SSSR 287:452-455 (1986), Adhin, MR. et al, Virology 170:238-242 (1989), Priano, C. et al, J. Mol. Biol. 249:283-297 (1995)). In particular the biological and biochemical properties of GA (Ni, CZ., et al, Protein Sci. 5:2485-2493 (1996), Tars, K et al, J. Mol.Biol. 271 :759-773(1997)) and of fr (Pushko P. et al, Prot. Eng. 6:883-891 (1993), Liljas, L et al. J Mol. Biol. 244:279-290, (1994)) have been disclosed. The crystal structure of several RNA bacteriophages has been determined (Golmohammadi, R. et al., Structure 4:543-554 (1996)).

In another preferred embodiment, the particle is a VLP of Cowpea cholortic mottle virus (CCMV). Assembly of CCMV virus from coat proteins expressed in E. Coli and nucleic acids has been described (Zhao X. et al. (1995) Virology 207:486-494).

In another embodiment, the particle is a VLP of Brome mosaic virus (BMV). Reassembly of BMV has been described previously (Choi YG and Rao LN (2000) Virology 275: 249-257, and references therein).

The immune-modulating substances in accordance with the present invention comprises, preferably consists of, INA, wherein preferably said INA is capable of inducing the production of IFN-alpha in a cell, preferably in a dendritic cell. In one embodiment, said INA is selected from (a) ribonucleic acids; (b) desoxyribonucleic acids, (c) chimeric nucleic acids; and (d) any mixtures of at least one nucleic acid of (a), (b) and/or (c). In another embodiment, said INA is a ribonucleic acid, preferably a double stranded ribonucleic acid, further preferably a double stranded ribonucleic acid selected from (a) double stranded viral RNA, or (b) synthetic double stranded RNA, preferably poly-(A:U) or poly(LC), again most preferably poly(LC). A preferred ribonucleic acid of the present invention comprises, preferably consists of polyinosinic- polycytidylic acid double-stranded RNA (poly I:C). In another embodiment said INA is a single stranded ribonucleic acid, preferably polyuridylic acid (poly-U, Westwood A. (2006), Vaccine 24: 1736-1743).

In another embodiment, said INA is an oligonucleotide, wherein said oligonucleotide is preferably selected from (a) unmethylated CpG-containing oligonucleotide; or (b) oligonucleotide free of unmethylated CpG motifs. Preferably, said INA is an unmethylated CpG- containing oligonucleotide.

Unmethylated CpG-dinucleotides within specific flanking bases represent important members of the microbial components that induce, modulate or enhance immune responses. Toll-like receptor 9 (TLR9) is activated by bacterial DNA, in particular by unmethylated CpG- containing oligonucleotides. In general, the unmethylated CpG-containing oligonucleotide comprises the sequence: 5' X1X2CGX3X4 3', wherein Xi, X₂, X3 and X₄ are any nucleotide. Specifically preferred oligonucleotides, unmethylated CpG-containing oligonucleotide, in the context of the invention comprise 20 to 40, preferably 26, 27, 28, 29, 30, 31 or 32 nucleotides, most preferably 30 nucleotides.

The CpG-containing oligonucleotide can contain one or more phosphothioester modifications of the phosphate backbone to enhance the stability of the oligonucleotide. CpG- containing oligonucleotides having one or more phosphate backbone modifications or having all of the phosphate backbone modified and CpG-containing oligonucleotides, wherein one, some or all of the nucleotide phosphate backbone modifications are phosphorothioate modifications are included within the scope of the present invention. In a preferred embodiment said INA is a CpG-containing oligonucleotide, wherein preferably said CpG-containing oligonucleotide consisting exclusively of phosphodiester bound, preferably unmethylated nucleotides are preferred in the context of the invention.

The CpG-containing oligonucleotide can also be recombinant, genomic, synthetic, cDNA, plasmid-derived and single or double stranded. For use in the nvention, the nucleic acids can be synthesized de novo using any of a number of procedures well known in the art. For example, the b-cyanoethyl phosphoramidite method (Beaucage, S. L., and Caruthers, M. H., Tet. Let. 22: 1859 (1981); nucleoside H-phosphonate method (Garegg et al, Tet. Let. 27:4051-4054 (1986); Froehler et al, Nucl. Acid. Res. 74:5399-5407 (1986); Garegg et al, Tet. Let. 27:4055- 4058 (1986), Gaffney et al, Tet. Let. 29:2619-2622 (1988)). These chemistries can be performed by a variety of automated oligonucleotide synthesizers available in the market. Alternatively, CpGs can be produced on a large scale in plasmids, (see Sambrook, T., et al., "Molecular Cloning: A Laboratory Manual," Cold Spring Harbor laboratory Press, New York, 1989) which after being administered to a patient are degraded into oligonucleotides. Oligonucleotides can be prepared from existing nucleic acid sequences (e.g., genomic or cDNA) using known techniques, such as those employing restriction enzymes, exonucleases or endonucleases.

In a very preferred embodiment said VLP, is packaged with said INA being a unmethylated CpG-containing oligonucleotide, most preferably G10 (SEQ ID NO: 6).

In another preferred embodiment of the present invention, the INA is an unmethylated CpG-containing oligonucleotide, wherein the CpG motif of said unmethylated CpG-containing oligonucleotide is part of a palindromic sequence. Preferably, said palindromic sequence is GACGATCGTC (SEQ ID NO: l).

In a preferred embodiment said INA is an A-type CpG or an C-type CpG. Preferably, said unmethylated CpG containing oligonucleotide is an A-type CpG, wherein preferably the nucleotides are exclusively linked by phosphodiester bonds. In a further preferred embodiment said INA is a A-type CpG comprising a palindromic sequence, wherein preferably said palindromic sequence is GACGATCGTC (SEQ ID NO: l).

In a preferred embodiment said palindromic sequence is flanked at its 5 '-terminus by at least 3 and at most 15, preferably at most 10, guanosine entities, wherein said palindromic sequence is flanked at its 3 '-terminus by at least 3 and at most 15, preferably at most 10 guanosine entities. In a further preferred embodiment, the palindromic sequence is flanked at its 5 '-terminus by at least 5 and at most 10 guanosine entities, and wherein said palindromic sequence is flanked at its 3 '-terminus by at least 5 and at most 10 guanosine entities. In a further preferred embodiment, the palindromic sequence, preferably SEQ ID NO:l, is flanked at its 3'- terminus by at least 10, preferably exactly 10, guanosine entities and at its 5 '-terminus by at least 10, preferably exactly 10, guanosine entities. In a very preferred embodiment said INA is a A- type CpG comprising a palindromic sequence, wherein said palindromic sequence is flanked at its 5 '-terminus by 3 to 10 guanosine entities, and wherein said palindromic sequence is flanked at its 3 '-terminus by 3 to 10 guanosine entities. In a even more preferred embodiment said INA is a A-type CpG comprising a palindromic sequence, wherein said palindromic is SEQ ID NO: l, and wherein said palindromic sequence is flanked at its 5 '-terminus by 3 to 10 guanosine entities, and wherein said palindromic sequence is flanked at its 3 '-terminus by 3 to 10 guanosine entities.

In a very preferred embodiment of the present invention, the unmethylated CpG-containing oligonucleotide comprises, or alternatively consists essentially of, or alternatively consists of the a sequence selected from the group consisting of (a) "G8-8" GGGGGGGG GACGATCGTC GGGGGGGG (SEQ ID NO:4); (b) "G9-9" GGGGGGGGG GACGATCGTC GGGGGGGGG (SEQ ID NO:5); or (c) "G10" GGGGGGGGGG GACGATCGTC GGGGGGGGGG (SEQ ID NO:6).

In a specifically preferred embodiment the unmethylated CpG-containing oligonucleotide comprises, or alternatively consists essentially of, or alternatively consists of "G10" (SEQ ID NO: 6), wherein preferably said G10 consists exclusively of phosphodiester bound nucleotides, wherein further preferably said G10 is aggregated G10 having a retention time relative to Q capsid standard under HPLC conditions, as set forth in Example 1 of WO2007/144150, of 50 to 110 %, most preferably of 80 to 110 %.

In a further specifically preferred embodiment the unmethylated CpG-containing oligonucleotide comprises, or alternatively consists essentially of, or alternatively consists of "G9-9" (SEQ ID NO:5). In a further specifically preferred embodiment the unmethylated CpG- containing oligonucleotide comprises, or alternatively consists essentially of, or alternatively consists of "G8-8" (SEQ ID NO:4).

In a further preferred embodiment of the present invention the INA is an unmethylated

CpG-containing oligonucleotide, wherein the CpG motif of said unmethylated CpG-containing oligonucleotide is part of a palindromic sequence, wherein said palindromic sequence is GACGATCGTC (SEQ ID NO:l), and wherein said palindromic sequence is flanked at its 5'- terminus of at least 4 and at most 9 guanosine entities and wherein said palindromic sequence is flanked at its 3 '-terminus of at least 6 and at most 9 guanosine entities.

In a further preferred embodiment of the present invention the ISS-NA is an unmethylated CpG-containing oligonucleotide, wherein the CpG motif of said unmethylated CpG-containing oligonucleotide is part of a palindromic sequence, wherein said palindromic sequence is GACGATCGTC (SEQ ID NO:21), and wherein said palindromic sequence is flanked at its 5'- terminus of at least 5 and at most 8 guanosine entities and wherein said palindromic sequence is flanked at its 3 '-terminus of at least 6 and at most 8 guanosine entities.

In a further preferred embodiment said INA is a unmethylated CpC containing oligonucleotide, wherein said unmethylated CpC containing oligonucleotide is capable of inducing the production of IFN-alpha in a cell, preferably in PBMCs, spleenocytes or human pDCs, and wherein further preferably said unmethylated CpC containing oligonucleotide is selected from: (a) T*C*G*T*C*G*T*T*T*T*G*T*C*G*T*T*T*T*G*T*C*G*T (2006-PS, SEQ ID NO: 9); (b) GGGGG AC GAT CGTCGGGGGG (2216-PO, SEQ ID NO: 10); (c) G*G*GGGACGATCGTC*G*G*G*G*G*G (2216-PO core, SEQ ID NO: 10); (d) GGTGCATCGATGCAGGGGGG (D19-PO, SEQ ID NO: 11); (e) G*G*TGCATCGATGCAG*G*G*G*G*G (D19-PO core, SEQ ID NO:l l), wherein * indicates a phosphorothioate modification, while all other nucleotides are phosphodiester bound.

In a more preferred embodiment said unmethylated CpG containing oligonucleotide is capable of inducing the production of IFN-alpha in human pDCs, wherein preferably said unmethylated CpG containing oligonucleotide is

_T*_C*_G*_T*_C*_G*_T*_T*_T=^ (2006-PS, SEQ ID NO: 9), wherein * indicates a phosphorothioate modification, while all other nucleotides are phosphodiester bound.

In a further preferred embodiment said ISS-NA is a C-type CpG, wherein preferably said C-type CpG comprises a palindromic sequence. In a further preferred embodiment said C-type CpG is SEQ ID NO: 12 or SEQ ID NO: 13, wherein preferably all nucleic acids of said C-type CpG are phosphorothioate bound. Further preferred C-type CpGs are selected from the group consisting of (a) TCpGTCGTTTT ACGGCGCCGTG CCG (SEQ ID NO: 12); (b) TCGTCGTTTT ACpGGCpGCCpGTGCCG (SEQ ID NO: 12); (c) TCGTCGTTT TACpGGCGCCpGTGCCG (SEQ ID NO: 12); and (d) TCGTCpGTTTT ACpGGCGCCpGTGCCG (SEQ ID NO: 12); wherein p indicates phosphodiester bounds while all other nucleotides are phosphorothioate bound.

C-type CpGs selected from the group consisting of (a) TCGTCGTTTTCGGCGCGCGCCG (SEQ ID NO: 14); (b) TCGTCGTTTTCGACGGCCGTCG (SEQ ID NO: 15); (c) TCGTCGTTTTCCGGCGCGCCGG (SEQ ID NO: 16); (d) TCGTCGTTTTCGGCGCGCGTCG (SEQ ID NO: 17); (e) TCGGCGCGCGCCGTCGTCGTTT (SEQ ID NO: 18); (f) TTGGCGCGCGCCGTCGTCGTTT (SEQ ID NO: 19); (g) TCGTCGTTTTCGTCGGCCGCCG (SEQ ID NO:20); (h) TCGTCGTTTTCGGCTTTTGCCG (SEQ ID NO:21); (i) TCGTCGTTTTCGGCGTTTTTTT (SEQ ID NO:22); and (j) TCGTCGTTTTCGGCGGCCGCCG (SEQ ID NO:23) are potent inducers of IFN-alpha production (Vollmer et al. 2004, Eur. J. Immunol. 43:351-262, p. 253, see Table 1 therein) and are thus specifically preferred ISS-NA in the context of the invention.

The invention also provides pharmaceutical compositions for use in a method for treating asthma of a patient in need thereof, said pharmaceutical composition comprising, essentially consisting of, or consisting of (a) a composition in accordance with the present discosure and/or invention; and (b) a pharmaceutically acceptable carrier.

As would be understood by one of ordinary skill in the art, when compositions of the invention are administered to an animal, they can be in a composition which contains salts, buffers, adjuvants or other substances which are desirable for improving the efficacy of the composition. Examples of materials suitable for use in preparing pharmaceutical compositions are provided in numerous sources including REMINGTON'S PHARMACEUTICAL SCIENCES (Osol, A, ed., Mack Publishing Co., (1990)). Compositions of the invention are said to be "pharmacologically acceptable" if their administration can be tolerated by a recipient individual.

The compositions of the present invention can be administered by various methods known in the art. The particular mode selected will depend of course, upon the particular composition selected, the severity of the asthma condition being treated and the dosage required for therapeutic efficacy, preferably to improve asthma control. The methods of the invention, generally speaking, can be practiced using any mode of administration that is medically acceptable, meaning any mode that produces effective levels without causing clinically unacceptable adverse effects. Such modes of administration include oral, rectal, parenteral, intracistemal, intravaginal, intraperitoneal, topical (as by powders, ointments, drops or transdermal patch), bucal, or as an oral or nasal spray.

Dosage levels depend on the mode of administration, the nature of the patient, and the quality of the carrier/adjuvant formulation. Typical amounts are in the range of about 0.001 μg to about 20 mg per patient. Preferred amounts are 50 μg to 1000 μg a composition of the invention per single administration. Multiple administrations to treat the patient is preferred. The administration of said composition or said pharmaceutical composition is repeated several times, preferably at least three to 10 times, most preferably three to five times, in daily, weekly, monthly or yearly intervals, preferably in intervals of about 1 week to about 1 month, or biweekly intervals.

A further aspect of the invention is a method of treating asthma of a patient in need thereof, wherein said method comprises (i) selecting a patient on controller treatment, wherein said patient is controlled by said patient's controller treatment, and wherein said patient having an eosinophilic cell count of more than 0.1 cells/nl in peripheral blood; and (ii) administering an effective amount of said composition to said patient, wherein said composition comprises, preferably consists of, (a) an inhaled glucocorticosteriod; (b) a long-acting p₂-agonist;(c) an immune-modulating substance; or (d) any mixture of at least one of (a), (b) and/or (c) wherein said administration is effective to improve asthma control of said patient.

In preferred embodiment, said method does not comprise introducing an allergen to said patient. In another embodiment an allergen is not introduced in said patient for at least one week before and at least one week after said introduction of said composition in said patient. In another embodiment, an allergen is not introduced in said patient for at least four weeks before and at least four weeks after said introduction of said composition in said patient. In another embodiment, an allergen is not introduced in said patient for at least eight weeks before and at least eight weeks after said introduction of said composition in said patient. In another preferred embodiment, said asthma is allergic asthma.

In all aspects and embodiments, in particular in all compositions, methods, and uses, of the invention, wherein said immune -modulating nucleic acid or said TLR9-ligand is the unmethylated CpG-containing oligonucleotide GIO (SEQ ID NO: 6) and wherein said particle is is a VLP of bacteriophage Q , said VLP of bacteriophage Q preferably consists of coat proteins comprising or, preferably consisting of the amino acid sequence of SEQ ID NO:7, and, further preferably, said unmethylated CpG-containing oligonucleotide GIO consists exclusively of phosphodiester bound nucleotides.

All patents, patent applications and publications referred to herein are expressly incorporated by reference in their entirety.

The pages 48 to 59 contain preferred embodiments of the present invention. Accordingly, the term "claim" as used within these pages 48 to 59 refers to "preferred embodiment".

1. A composition for use in a method for treating asthma of a patient in need thereof, wherein said method comprises

(i) selecting a patient on controller treatment, wherein said patient is controlled by said patient's controller treatment, and wherein said patient has an eosinophilic cell count of greater than 0.1 cells/nl in peripheral blood; and

(ii) administering an effective amount of said composition to said patient, wherein said composition comprises, preferably consists of,

(a) an inhaled glucocorticosteriod;

(b) a long-acting p₂-agonist;

(c) an immune -modulating substance; or

(d) any mixture of at least one of (a), (b) and/or (c)

wherein said administration is effective to improve asthma control of said patient.

2. The composition of claim 1, wherein said patient has an eosinophilic cell count of equal to or greater than 0.15 cells/nl in peripheral blood, and wherein preferably said patient has an eosinophilic cell count of greater than 0.15 cells/nl in peripheral blood .

3. The composition of claim 1 or 2, wherein said patient has an eosinophilic cell count of less than 0.45 cells/nl in peripheral blood.

4. The composition of claim 1 or 2, wherein said patient has an eosinophilic cell count of equal to or less than 0.4 cells/nl in peripheral blood.

5. The composition of claim 1 or 2, wherein said patient has an eosinophilic cell count of less than 0.25 cells/nl in peripheral blood.

6. The composition of claim 1 or 2, wherein said patient has an eosinophilic cell count of less than 0.3 cells/nl in peripheral blood.

7. The composition of any one of the preceding claims, wherein said patient's controller treatment is a treatment of GINA Step 2.

8. The composition of any one of the preceding claims, wherein said patient's controller treatment is a treatment of GINA Step 3.

9. The composition of any one of the preceding claims, wherein said patient's controller treatment is treatment by way of inhaled glucocorticosteriods. The composition of any one of the preceding claims, wherein said patient's controller treatment is treatment by way of low dose inhaled glucocorticosteriods.

The composition of any one of the preceding claims, wherein said patient's controller treatment is treatment by way of medium dose inhaled glucocorticosteriods.

The composition of any one of the preceding claims, wherein said patient's controller treatment is treatment by way of high dose inhaled glucocorticosteriods.

The composition of any one of the preceding claims, wherein said patient's controller treatment is a treatment of GINA Step 4.

The composition of any one of the claims 1 to 6, 8 or 13, wherein said patient's controller treatment is treatment by way of inhaled corticosteroids combined with long-acting β₂- agonists.

The composition of claim 14, wherein said patient's controller treatment is treatment by way of low dose inhaled glucocorticosteriods combined with long-acting ₂-agonists.

The composition of claim 14 or claim 15, wherein said patient's controller treatment is treatment by way of medium dose inhaled glucocorticosteriods combined with long-acting 2-agonists.

The composition of any one of the claims 14 to 16, wherein said patient's controller treatment is treatment by way of high dose inhaled glucocorticosteriods combined with long-acting ₂-agonists.

The composition of any one of the preceding claims, wherein said administering of said effective amount of said composition to said patient is repeated at least once, at least twice, at least three, four, five, six, or seven times, or at least ten times.

The composition of any one of the preceding claims, wherein said administering of said effective amount of said composition to said patient comprises repeated administrations of said effective amount of said composition to said patient, wherein said repeated administrations are effected in intervals of 1 week, 2 weeks, 3 weeks, monthly, bimonthly, quarterly or semi-annually.

The composition of any one of the preceding claims, wherein said patient's controller treatment is continued during said administering said effective amount of said composition to said patient.

The composition of any one of the claims 1 to 19, wherein said patient's controller treatment is reduced during said administering said effective amount of said composition to said patient. The composition of claim 21, wherein said reduction of said patient's controller treatment is effected at least 12 weeks, preferably at least 8 weeks, more preferably at least 4 weeks after a first administration of said effective amount of said composition.

The composition of any one of the claims 1 to 19, wherein said patient's controller treatment is removed during said administering said effective amount of said composition to said patient.

The composition of claim 23, wherein said removal of said patient's controller treatment is effected at least 12 weeks, preferably at least 8 weeks, more preferably at least 4 weeks after a first administration of said effective amount of said composition

The composition of any one of the preceding claims, wherein said administration is effective to improve asthma control of said patient as measured by either a reduction of ACQ-7 of at least of 0.3, preferably 0.5 points.

The composition of any one of the preceding claims, wherein said composition comprises, preferably consists of, an inhaled glucocorticosteriod.

The composition of any one of the preceding claims, wherein said effective amount of said inhaled glucocorticosteriod is a low dose inhaled glucocorticosteriod.

The composition of any one of the preceding claims, wherein said effective amount of said inhaled glucocorticosteriod is a medium dose inhaled glucocorticosteriod.

The composition of any one of the preceding claims, wherein said effective amount of said inhaled glucocorticosteriod is a high dose inhaled glucocorticosteriod.

The composition of any one of the claims 1 to 25, wherein said composition comprises, preferably consists of, a combination of an inhaled glucocorticosteriod with a long-acting p₂-agonist.

The composition of claim 30, wherein said effective amount of said composition is a combination of a low dose inhaled glucocorticosteriod with a long-acting ₂-agonist The composition of claim 30, wherein said effective amount of said composition is a combination of a medium dose inhaled glucocorticosteriod with a long-acting ₂-agonist. The composition of claim 30, wherein said effective amount of said composition is a combination of a high dose inhaled glucocorticosteriod with a long-acting ₂-agonist. The composition of any one of the preceding claims, wherein said composition comprises, preferably consists of, an immune -modulating substance, wherein preferably said immune- modulating substance comprises, preferably consists of,

(i) an immune-modulating nucleic acid;

(ii) a toll-like receptor 7 ligand (TLR7-ligand);

(iii) a toll-like receptor 9 (TLR9-ligand); (iv) a chemoattractant receptor-homologous molecule expressed on Th2

cells antagonist (CRTh2 anatagonist);

(v) a chemokine antagonist;

(vi) a somatotaxin; or

(vii) any mixture of at least one of (i), (ii), (iii), (iv), (v) and/or (vi).

The composition of any one of the preceding claims, wherein said composition comprises, preferably consists of, an immune-modulating substance, wherein said immune -modulating substance comprises, preferably consists of:

(i) an immune-modulating nucleic acid;

(ii) a TLR7-ligand;

(iii) a TLR9-ligand; or

(iv) any mixture of at least one of (i), (ii) and/or (iii).

The composition of any one of the preceding claims, wherein said composition comprises, preferably consists of, an immune-modulating substance, wherein said immune -modulating substance comprises, preferably consists of, an immune-modulating nucleic acid.

The composition of claim 36, wherein said immune-modulating nucleic acid is capable of stimulating IFN-alpha production in a cell.

The composition of claim 36 or claim 37, wherein said immune-modulating nucleic acid is selected from:

(a) a desoxyribonucleic acid;

(b) a ribonucleic acid,

(c) a chimeric nucleic acid; or

(d) any mixture of at least one nucleic acid of (a), (b) and/or (c).

The composition of any one of claims 36 to 38, wherein said immune-modulating nucleic acid comprises, preferably consists of, an unmethylated CpG containing oligonucleotide. The composition of claim 38, wherein said ribonucleic acid comprises, preferably consists of, poly-(LC) or a derivative thereof.

The composition of claim 38, wherein said deoxyribonucleic acid comprises, preferably consists of, an oligonucleotide free of an unmethylated CpG motif.

The composition of any one of the claims 1 to 35, wherein said composition comprises, preferably consists of, an immune-modulating substance, wherein said immune -modulating substance comprises, preferably consists of, a TLR7-ligand. The composition of any one of the claims 1 to 35, wherein said composition comprises, preferably consists of, an immune-modulating substance, wherein said immune -modulating substance comprises, preferably consists of, a TLR9-ligand.

The composition of claim 43, wherein said TLR9-ligand comprises, preferably consists of, an unmethylated CpG containing oligonucleotide.

The composition of any one of claims 1 to 33, wherein said composition comprises, preferably consists of, an immune-modulating substance, wherein said immune -modulating substance comprises, preferably consists of an unmethylated CpG containing oligonucleotide.

The composition of any one of the claims 39, 44 or 45, wherein said unmethylated CpG containing oligonucleotide is selected from of:

(a) A-type CpGs;

(b) B-type CpGs; or

(c) C-type CpGs.

The composition of any one of the claims 39, 44 or 45, wherein said unmethylated CpG- containing oligonucleotide is an A-type CpG.

The composition of any one of the claims 39, 44 or 45, wherein said unmethylated CpG- containing oligonucleotide is a B-type CpG.

The composition of any one of the claims 39, 44 or 45, wherein said unmethylated CpG- containing oligonucleotide is a C-type CpG.

The composition of any one of the claims 39, 44 or 45 to 49, wherein said unmethylated CpG-containing oligonucleotide comprises a palindromic sequence.

The composition of any one of the claims 39, 44 or 45 to 49, wherein the CpG motif of said unmethylated CpG-containing oligonucleotide is part of a palindromic sequence. The composition of claim 50 or 51, wherein said palindromic sequence is GACGATCGTC (SEQ ID NO: l).

The composition of any one of claims 50 to 52, wherein said palindromic sequence is flanked at its 5 '-terminus and at its 3 '-terminus by guanosine entities.

The composition of any one of claims 50 to 53, wherein said palindromic sequence is flanked at its 5 '-terminus by at least 3 and at most 15 guanosine entities, and wherein said palindromic sequence is flanked at its 3 '-terminus by at least 3 and at most 15 guanosine entities. 55. The composition of any one of the claims 39, 44 or 45 to 54, wherein said unmethylated CpG-containing oligonucleotide comprises or alternatively consists of the sequence selected from:

(a) "G6-6" GGGGGGGACGATCGTCGGGGGG (SEQ ID NO:2);

(b) "G7-7" GGGGGGGGACGATCGTCGGGGGGG (SEQ ID NO:3);

(c) "G8-8" GGGGGGGGGACGATCGTCGGGGGGGG (SEQ ID NO:4);

(d) "G9-9" GGGGGGGGGGACGATCGTCGGGGGGGGG (SEQ ID NO:5); or

(e) "G10" GGGGGGGGGGGACGATCGTCGGGGGGGGGG (SEQ ID NO:6).

56. The composition of any one of the claims 39, 44 or 45 to 55, wherein said unmethylated CpG-containing oligonucleotide comprises, or alternatively consists essentially of, or alternatively consists of the sequence GGGGGGGGGGGACGATCGTCGGGGGGGGGG (SEQ ID NO:6).

57. The composition of any one of the claims 39, 44 or 45 to 56, wherein said unmethylated CpG-containing oligonucleotide consists exclusively of phosphodiester bound nucleotides. 58. The composition of any one of the claims 39, 44 or 45 to 57, wherein said immune- modulating nucleic acid, preferably said unmethylated CpG-containing oligonucleotide, is not accessible to DNAse hydrolysis.

59. The composition of any one of the claims 39, 44 or 45 to 58, wherein said immune- modulating nucleic acid is an unmethylated CpG-containing oligonucleotide, and wherein said unmethylated CpG-containing oligonucleotide is not accessible to Benzonase hydrolysis.

60. The composition of any one of the claims 39, 44 or 45 to 59, wherein said immune- modulating nucleic acid is an unmethylated CpG containing oligonucleotide consisting of the sequence GGGGGGGGGGGACGATCGTCGGGGGGGGGG (SEQ ID NO:6), and wherein said unmethylated CpG-containing oligonucleotide consists exclusively of phosphodiester bound nucleotides.

61. The composition of any one of the claims 1 to 33, wherein said composition comprises, preferably consists of, an immune-modulating substance, wherein said immune -modulating substance comprises, preferably consists of:

(i) an immune-modulating nucleic acid;

(ii) a TLR7-ligand;

(iii) a TLR9-ligand; or

(iv) any mixture of at least one of (i), (ii) and/or (iii);

wherein said immune-modulating substance is packaged in a particle, wherein said particle is, preferably, a virus-like particle or a virosome. The composition of claim 61, wherein said particle is a virus-like particle.

The composition of claim 61, wherein said virus-like particle is a virus-like particle of a bacteriophage.

The composition of claim 61, wherein said virus-like particle is a virus-like particle of a RNA bacteriophage.

The composition of any one of claims 62 to 64, wherein said virus-like particle comprises, or alternatively essentially consists of, alternatively consists of recombinant proteins, or fragments thereof, of a RNA bacteriophage.

The composition of any one of claims 62 to 64, wherein said virus-like particle comprises, or alternatively essentially consists of, alternatively consists of coat proteins, or fragments thereof, of a RNA bacteriophage.

The composition of any one of claims 64 to 66, wherein said RNA bacteriophage is selected from:

(a) bacteriophage QP;

(b) bacteriophage R17;

(c) bacteriophage fr;

(d) bacteriophage GA;

(e) bacteriophage SP;

(f) bacteriophage MS2;

(g) bacteriophage Mi l;

CO bacteriophage MX1;

(i) bacteriophage NL95;

0) bacteriophage f2;

(k) bacteriophage PP7; or

(1) bacteriophage AP205.

The composition of any one of claims 64 to 66, wherein said RNA bacteriophage is QP. The composition of claim 62, wherein said virus-like particle is a virus-like particle of a RNA bacteriophage QP.

The composition of claim 69, wherein said virus-like particle of a RNA bacteriophage QP consists of coat proteins having the amino acid sequence of SEQ ID NO:7.

The composition of claim 69, wherein said virus-like particle of a RNA bacteriophage QP consists of coat proteins consisting of the amino acid sequence of SEQ ID NO:7. 72. The composition of any one of claims 64 to 66, wherein said R A bacteriophage is AP205.

73. The composition of any one claims to, 61 to 72, wherein said immune -modulating substance comprises, preferably consists of, an immune-modulating nucleic acid.

74. The composition of claim 73, wherein said immune-modulating nucleic acid is capable of stimulating IFN-alpha production in a cell.

75. The composition of claim 73 or claim 74, wherein said immune-modulating nucleic acid is selected from:

(a) a desoxyribonucleic acid;

(b) a ribonucleic acid ,

(c) a chimeric nucleic acid; or

(d) any mixture of at least one nucleic acid of (a), (b) and/or (c).

76. The composition of any one of claims 73 to 75, wherein said immune-modulating nucleic acid comprises, preferably consists of, an unmethylated CpG containing oligonucleotide.

77. The composition of claim 75, wherein said ribonucleic acid comprises, preferably consists of, poly-(LC) or a derivative thereof.

78. The composition of claim 75, wherein said deoxyribonucleic acid comprises, preferably consists of, an oligonucleotide free of an unmethylated CpG motif.

79. The composition of any one claims to, 61 to 73, wherein said immune -modulating substance comprises, preferably consists of, a TLR7-ligand.

80. The composition of any one claims to, 61 to 73, wherein said said immune -modulating substance comprises, preferably consists of, a TLR9-ligand.

81. The composition of claim 80, wherein said TLR9-ligand comprises, preferably consists of, an unmethylated CpG containing oligonucleotide.

82. The composition of claim 81, wherein said unmethylated CpG containing oligonucleotide is selected from of:

(a) A-type CpGs;

(b) B-type CpGs; or

(b) C-type CpGs.

83. The composition of claim 81, wherein said unmethylated CpG-containing oligonucleotide is an A-type CpG.

84. The composition of claim 81, wherein said unmethylated CpG-containing oligonucleotide is a B-type CpG. 85. The composition of claim 81, wherein said unmethylated CpG-containing oligonucleotide is a C-type CpG.

86. The composition of claim 81 to 85, wherein said unmethylated CpG-containing oligonucleotide comprises a palindromic sequence.

87. The composition of claim 81 to 86, wherein the CpG motif of said unmethylated CpG- containing oligonucleotide is part of a palindromic sequence.

88. The composition of claim 86 or 87, wherein said palindromic sequence is GACGATCGTC (SEQ ID NO: l).

89. The composition of any one of claims 86 to 88, wherein said palindromic sequence is flanked at its 5 '-terminus and at its 3 '-terminus by guanosine entities.

90. The composition of any one of claims 86 to 89, wherein said palindromic sequence is flanked at its 5 '-terminus by at least 3 and at most 15 guanosine entities, and wherein said palindromic sequence is flanked at its 3 '-terminus by at least 3 and at most 15 guanosine entities.

91. The composition of claim 81 to 90, wherein said unmethylated CpG-containing oligonucleotide comprises or alternatively consists of the sequence selected from the group consisting of:

(a) "G6-6" GGGGGGGACGATCGTCGGGGGG (SEQ ID NO:2);

(b) "G7-7" GGGGGGGGACGATCGTCGGGGGGG (SEQ ID NO:3);

(c) "G8-8" GGGGGGGGGACGATCGTCGGGGGGGG (SEQ ID NO:4);

(d) "G9-9" GGGGGGGGGGACGATCGTCGGGGGGGGG (SEQ ID NO:5); and

(e) "G10" GGGGGGGGGGGACGATCGTCGGGGGGGGGG (SEQ ID NO:6).

92. The composition of claim 81 to 91, wherein said unmethylated CpG-containing oligonucleotide comprises, or alternatively consists essentially of, or alternatively consists of the sequence GGGGGGGGGGGACGATCGTCGGGGGGGGGG (SEQ ID NO:6).

93. The composition of claim 81 to 92, wherein said unmethylated CpG-containing oligonucleotide consists exclusively of phosphodiester bound nucleotides.

94. The composition of claim 81 to 93, wherein said immune-modulating nucleic acid, preferably said unmethylated CpG-containing oligonucleotide, is not accessible to DNAse hydrolysis.

95. The composition of claim 81 to 94, wherein said immune-modulating nucleic acid is an unmethylated CpG-containing oligonucleotide, and wherein said unmethylated CpG- containing oligonucleotide is not accessibly to Benzonase hydrolysis. 96. The composition of claim 81 to 95, wherein said immune-modulating nucleic acid is an unmethylated CpG containing oligonucleotide consisting of the sequence GGGGGGGGGGGACGATCGTCGGGGGGGGGG (SEQ ID NO: 6), and wherein said unmethylated CpG-containing oligonucleotide consists exclusively of phosphodiester bound nucleotides.

97. The composition of any one of the preceding claims, wherein said administering said effective amount of said composition to said patient is repeated at least once, preferably at least twice, more preferably at least three, four or five times, again further preferably at least six times, and again further preferably six times, and wherein said repeated administration of said composition to said patient is effected in intervals of 1 week or 2 weeks.

98. The composition of any one of the preceding claims, wherein said administering said effective amount of said composition to said patient is effected three times in intervals of 1 week followed by four times in intervals of 2 weeks.

99. The composition of any one of the preceding claims, wherein said effective amount of said composition is 900 μg.

100. The composition of any one of the preceding claims, wherein said composition is administered to said patient subcutaneously, intramuscularly, intravenously, intranasally or directly into the lymph node.

101. The composition of any one of the preceding claims, wherein said composition is administered to said patient subcutaneously.

102. The composition of any one of the preceding claims, wherein said composition is administered to said patient without an allergen.

103. The composition of any one of the preceding claims, wherein said composition is administered to said patient without administration of an allergen.

104. The composition of any one of the preceding claims, wherein said composition is not administered to said patient in conjunction with an allergen.

105. The composition of any one of the preceding claims, wherein said method does not comprise co-administering an allergen to said patient.

106. The composition of any one of the preceding claims, wherein said composition does not comprise an allergen.

107. The composition of any one of the preceding claims, wherein said method does not comprise administering an allergen to said patient. 108. The composition of any one of the preceding claims, wherein an allergen is not introduced in said patient for at least one, preferably for at least four, and further preferably for at least eight weeks before and at least one week, preferably for at least four weeks, and further preferably for at least eight weeks after said administering of said effective amount of said composition to said patient.

109. The composition of any one of the preceding claims, wherein said composition is comprised by a pharmaceutical composition, wherein said pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

110. The composition of any one of the preceding claims, wherein said asthma is allergic asthma.

111. A pharmaceutical composition for use in a method for treating asthma of a patient in need thereof, said pharmaceutical composition comprising, essentially consisting of, or consisting of:

(a) a composition of any one of claims 1 to 110; and

(b) a pharmaceutically acceptable carrier.

112. A method of treating asthma of a patient in need thereof, wherein said method comprises

(i) selecting a patient on controller treatment, wherein said patient is controlled by said patient's controller treatment, and wherein said patient having an eosinophilic cell count of more than 0.1 cells/nl in peripheral blood; and

(ii) administering an effective amount of said composition to said patient, wherein said composition comprises, preferably consists of,

(a) an inhaled glucocorticosteriod;

(b) a long-acting p₂-agonist;

(c) an immune-modulating substance; or

(d) any mixture of at least one of (a), (b) and/or (c)

wherein said administration is effective to improve asthma control of said patient.

113. The method of claim 112, wherein said method does not comprise introducing an allergen to said patient.

114. The method of claim 1 12 or claim 113, wherein an allergen is not introduced in said patient for at least one week before and at least one week after said introduction of said composition in said patient.

115. The method of any one of the claims 112 to 114, wherein an allergen is not introduced in said patient for at least four weeks before and at least four weeks after said introduction of said composition in said patient. 116. The method of any one of the claims 112 to 115, wherein an allergen is not introduced in said patient for at least eight weeks before and at least eight weeks after said introduction of said composition in said patient.

117. The method of any one of the claims 112 to 116, wherein the composition for use is defined as in any one of the claims 1 to 110.

118. The method of any one of the claims 112 to 117, wherein said asthma is allergic asthma.

In the context of the present invention, the following sequence identifiers (SEQ ID NO's) have been assigned to the following sequences:

SEQ ID NO:l gacgatcgtc

SEQ ID NO: 2 gggggggacg atcgtcgggg gg

SEQ ID NO: 3 ggggggggac gatcgtcggg gggg

SEQ ID NO: 4 ggggggggga cgatcgtcgg gggggg

SEQ ID NO: 5 gggggggggg acgatcgtcg gggggggg

SEQ ID NO: 6 gggggggggg gacgatcgtc gggggggggg

SEQ ID NO: 7 Ala Lys Leu Glu Thr Val Thr Leu Gly Asn He Gly Lys Asp

Gly Lys Gin Thr Leu Val Leu Asn Pro Arg Gly Val Asn Pro

Thr Asn Gly Val Ala Ser Leu Ser Gin Ala Gly Ala Val Pro

Ala Leu Glu Lys Arg Val Thr Val Ser Val Ser Gin Pro Ser

Arg Asn Arg Lys Asn Tyr Lys Val Gin Val Lys He Gin Asn

Pro Thr Ala Cys Thr Ala Asn Gly Ser Cys Asp Pro Ser Val

Thr Arg Gin Ala Tyr Ala Asp Val Thr Phe Ser Phe Thr Gin

Tyr Ser Thr Asp Glu Glu Arg Ala Phe Val Arg Thr Glu Leu

Ala Ala Leu Leu Ala Ser Pro Leu Leu He Asp Ala He Asp

Gin Leu Asn Pro Ala Tyr

SEQ ID NO: Met Ala Lys Leu Glu Thr Val Thr Leu Gly Asn He Gly Lys

Asp Gly Lys Gin Thr Leu Val Leu Asn Pro Arg Gly Val Asn

Pro Thr Asn Gly Val Ala Ser Leu Ser Gin Ala Gly Ala Val

Pro Ala Leu Glu Lys Arg Val Thr Val Ser Val Ser Gin Pro

Ser Arg Asn Arg Lys Asn Tyr Lys Val Gin Val Lys He Gin

Asn Pro Thr Ala Cys Thr Ala Asn Gly Ser Cys Asp Pro Ser

Val Thr Arg Gin Ala Tyr Ala Asp Val Thr Phe Ser Phe Thr

Gin Tyr Ser Thr Asp Glu Glu Arg Ala Phe Val Arg Thr Glu

Leu Ala Ala Leu Leu Ala Ser Pro Leu Leu He Asp Ala He

Asp Gin Leu Asn Pro Ala Tyr Trp Thr Leu Leu He Ala Gly

Gly Gly Ser Gly Ser Lys Pro Asp Pro Val He Pro Asp Pro

Pro He Asp Pro Pro Pro Gly Thr Gly Lys Tyr Thr Cys Pro

Phe Ala He Trp Ser Leu Glu Glu Val Tyr Glu Pro Pro Thr

Lys Asn Arg Pro Trp Pro He Tyr Asn Ala Val Glu Leu Gin

Pro Arg Glu Phe Asp Val Ala Leu Lys Asp Leu Leu Gly Asn

Thr Lys Trp Arg Asp Trp Asp Ser Arg Leu Ser Tyr Thr Thr

Phe Arg Gly Cys Arg Gly Asn Gly Tyr He Asp Leu Asp Ala

Thr Tyr Leu Ala Thr Asp Gin Ala Met Arg Asp Gin Lys Tyr

Asp He Arg Glu Gly Lys Lys Pro Gly Ala Phe Gly Asn He

Glu Arg Phe He Tyr Leu Lys Ser He Asn Ala Tyr Cys Ser

Leu Ser Asp He Ala Ala Tyr His Ala Asp Gly Val He Val

Gly Phe Trp Arg Asp Pro Ser Ser Gly Gly Ala He Pro Phe

Asp Phe Thr Lys Phe Asp Lys Thr Lys Cys Pro He Gin Ala

Val He Val Val Pro Arg Ala

SEQ ID NO: 9 tcgtcgtttt gtcgttttgt cgt

SEQ ID NO:10 gggggacgat cgtcgggggg

SEQ ID NO:ll ggtgcatcga tgcagggggg

SEQ ID NO:12 tcgtcgtttt acggcgccgt gccg

SEQ ID NO:13 tcgtcgtttt acggcgtcgt gccg

SEQ ID NO:14 tcgtcgtttt cggcgcgcgc eg SEQ ID NO:15 tcgtcgtttt cgacggccgt eg

SEQ ID NO:16 tcgtcgtttt ccggcgcgcc gg

SEQ ID NO: 17 tcgtcgtttt cggcgcgcgt eg

SEQ ID NO:18 tcggcgcgcg ccgtcgtcgt tt

SEQ ID NO:19 ttggcgcgcg ccgtcgtcgt tt

SEQ ID NO:20 tcgtcgtttt cgtcggccgc eg

SEQ ID N0:21 tcgtcgtttt eggcttttge eg

SEQ ID NO:22 tcgtcgtttt cggcgttttt tt

SEQ ID NO:23 tcgtcgtttt cggcggccgc eg

In a further very preferred embodiment, the present invention provides a composition for use in a method for treating asthma of a patient in need thereof, wherein said method comprises (i) selecting a patient on controller treatment, wherein said patient is controlled by said patient's controller treatment, and wherein said patient has an eosinophilic cell count of greater than 0.1 cells/nl in peripheral blood, preferably equal to or greater than 0.15 cells/nl; and (ii) administering an effective amount of said composition to said patient, wherein said composition is an immune-modulating substance, wherein said immune-modulating substance is packaged in a virus-like particle of a R A bacteriophage Q consisting of coat proteins consisting of the amino acid sequence of SEQ ID NO:7, and wherein said immune-modulating substance consists of an unmethylated CpG containing oligonucleotide consisting of the sequence GGGGGGGGGGGACGATCGTCGGGGGGGGGG (SEQ ID NO: 6), and wherein said unmethylated CpG-containing oligonucleotide consists exclusively of phosphodiester bound nucleotides; and wherein said administration is effective to improve asthma control of said patient.

In a further very preferred embodiment, the present invention provides a composition for use in a method for treating asthma of a patient in need thereof, wherein said method comprises (i) selecting a patient on controller treatment, wherein said patient is controlled by said patient's controller treatment, and wherein said patient has an eosinophilic cell count of greater than 0.1 cells/nl and less than 0.3 cells/nl in peripheral blood preferably of greater than 0.15 cells/nl and less than 0.25 cells/nl; and (ii) administering an effective amount of said composition to said patient, wherein said composition is an immune-modulating substance, wherein said immune- modulating substance is packaged in a virus-like particle of a RNA bacteriophage Q consisting of coat proteins consisting of the amino acid sequence of SEQ ID NO:7, and wherein said immune -modulating substance consists of an unmethylated CpG containing oligonucleotide consisting of the sequence GGGGGGGGGGGACGATCGTCGGGGGGGGGG (SEQ ID NO:6), and wherein said unmethylated CpG-containing oligonucleotide consists exclusively of phosphodiester bound nucleotides; and wherein said administration is effective to improve asthma control of said patient.

In a further very preferred embodiment, the present invention provides a composition for use in a method for treating asthma of a patient in need thereof, wherein said asthma is allergic asthma, wherein said method comprises (i) selecting a patient on controller treatment, wherein said patient is controlled by said patient's controller treatment, and wherein said patient has an eosinophilic cell count of greater than 0.1 cells/nl in peripheral blood, preferably equal to or greater than 0.15 cells/nl; and (ii) administering an effective amount of said composition to said patient, wherein said composition is an immune -modulating substance, wherein said immune- modulating substance is packaged in a virus-like particle of a R A bacteriophage Q consisting of coat proteins consisting of the amino acid sequence of SEQ ID NO:7, and wherein said immune -modulating substance consists of an unmethylated CpG containing oligonucleotide consisting of the sequence GGGGGGGGGGGACGATCGTCGGGGGGGGGG (SEQ ID NO:6), and wherein said unmethylated CpG-containing oligonucleotide consists exclusively of phosphodiester bound nucleotides; and wherein preferably said composition does not comprise an allergen and is not administered to said patient in conjunction with an allergen, and wherein said administration is effective to improve asthma control of said patient, and wherein preferably said method does not comprise co-administering an allergen to said patient.

In a further very preferred embodiment, the present invention provides a composition for use in a method for treating asthma of a patient in need thereof, wherein said asthma is allergic asthma, and wherein said method comprises (i) selecting a patient on controller treatment, wherein said patient is controlled by said patient's controller treatment, and wherein said patient has an eosinophilic cell count of greater than 0.1 cells/nl and less than 0.3 cells/nl in peripheral blood preferably of greater than 0.15 cells/nl and less than 0.25 cells/nl; and (ii) administering an effective amount of said composition to said patient, wherein said composition is an immune- modulating substance, wherein said immune-modulating substance is packaged in a virus-like particle of a RNA bacteriophage Q consisting of coat proteins consisting of the amino acid sequence of SEQ ID NO:7, and wherein said immune-modulating substance consists of an unmethylated CpG containing oligonucleotide consisting of the sequence GGGGGGGGGGGACGATCGTCGGGGGGGGGG (SEQ ID NO: 6), and wherein said unmethylated CpG-containing oligonucleotide consists exclusively of phosphodiester bound nucleotides, and wherein preferably said composition does not comprise an allergen and is not administered to said patient in conjunction with an allergen; and wherein said administration is effective to improve asthma control of said patient, and wherein preferably said method does not comprise co-administering an allergen to said patient.

In a further very preferred embodiment, the present invention provides a composition for use in a method for treating asthma of a patient in need thereof, wherein said method comprises (i) selecting a patient on controller treatment, wherein said patient is controlled by said patient's controller treatment, and wherein said patient has an eosinophilic cell count of greater than 0.1 cells/nl in peripheral blood, preferably equal to or greater than 0.15 cells/nl; and (ii) administering an effective amount of said composition to said patient, wherein said composition is an immune-modulating substance, wherein said immune-modulating substance is packaged in a virus-like particle of a R A bacteriophage Q consisting of coat proteins consisting of the amino acid sequence of SEQ ID NO:7, and wherein said immune-modulating substance consists of an immune-modulating nucleic acid, and wherein said immune-modulating nucleic acid is an unmethylated CpG containing oligonucleotide consisting of the sequence GGGGGGGGGGGACGATCGTCGGGGGGGGGG (SEQ ID NO: 6), and wherein said unmethylated CpG-containing oligonucleotide consists exclusively of phosphodiester bound nucleotides; and wherein said administration is effective to improve asthma control of said patient.

In a further very preferred embodiment, the present invention provides a composition for use in a method for treating asthma of a patient in need thereof, wherein said method comprises (i) selecting a patient on controller treatment, wherein said patient is controlled by said patient's controller treatment, and wherein said patient has an eosinophilic cell count of greater than 0.1 cells/nl and less than 0.3 cells/nl in peripheral blood preferably of greater than 0.15 cells/nl and less than 0.25 cells/nl; and (ii) administering an effective amount of said composition to said patient, wherein said composition is an immune-modulating substance, wherein said immune- modulating substance is packaged in a virus-like particle of a RNA bacteriophage Q consisting of coat proteins consisting of the amino acid sequence of SEQ ID NO:7, and wherein said immune-modulating substance consists of an immune-modulating nucleic acid, and wherein said immune -modulating nucleic acid is an unmethylated CpG containing oligonucleotide consisting of the sequence GGGGGGGGGGGACGATCGTCGGGGGGGGGG (SEQ ID NO:6), and wherein said unmethylated CpG-containing oligonucleotide consists exclusively of phosphodiester bound nucleotides; and wherein said administration is effective to improve asthma control of said patient.

In a further very preferred embodiment, the present invention provides a composition for use in a method for treating asthma of a patient in need thereof, wherein said asthma is allergic asthma, wherein said method comprises (i) selecting a patient on controller treatment, wherein said patient is controlled by said patient's controller treatment, and wherein said patient has an eosinophilic cell count of greater than 0.1 cells/nl in peripheral blood, preferably equal to or greater than 0.15 cells/nl; and (ii) administering an effective amount of said composition to said patient, wherein said composition is an immune -modulating substance, wherein said immune - modulating substance is packaged in a virus-like particle of a R A bacteriophage Q consisting of coat proteins consisting of the amino acid sequence of SEQ ID NO:7, and wherein said immune-modulating substance consists of an immune-modulating nucleic acid, and wherein said immune -modulating nucleic acid is an unmethylated CpG containing oligonucleotide consisting of the sequence GGGGGGGGGGGACGATCGTCGGGGGGGGGG (SEQ ID NO:6), and wherein said unmethylated CpG-containing oligonucleotide consists exclusively of phosphodiester bound nucleotides; and wherein preferably said composition does not comprise an allergen and is not administered to said patient in conjunction with an allergen, and wherein said administration is effective to improve asthma control of said patient, and wherein preferably said method does not comprise co-administering an allergen to said patient.

In a further very preferred embodiment, the present invention provides a composition for use in a method for treating asthma of a patient in need thereof, wherein said asthma is allergic asthma, and wherein said method comprises (i) selecting a patient on controller treatment, wherein said patient is controlled by said patient's controller treatment, and wherein said patient has an eosinophilic cell count of greater than 0.1 cells/nl and less than 0.3 cells/nl in peripheral blood preferably of greater than 0.15 cells/nl and less than 0.25 cells/nl; and (ii) administering an effective amount of said composition to said patient, wherein said composition is an immune- modulating substance, wherein said immune-modulating substance is packaged in a virus-like particle of a RNA bacteriophage Q consisting of coat proteins consisting of the amino acid sequence of SEQ ID NO:7, and wherein said immune-modulating substance consists of an immune-modulating nucleic acid, and wherein said immune-modulating nucleic acid is an unmethylated CpG containing oligonucleotide consisting of the sequence GGGGGGGGGGGACGATCGTCGGGGGGGGGG (SEQ ID NO: 6), and wherein said unmethylated CpG-containing oligonucleotide consists exclusively of phosphodiester bound nucleotides, and wherein preferably said composition does not comprise an allergen and is not administered to said patient in conjunction with an allergen; and wherein said administration is effective to improve asthma control of said patient, and wherein preferably said method does not comprise co-administering an allergen to said patient.

In a further very preferred embodiment, the present invention provides a composition for use in a method for treating asthma of a patient in need thereof, and wherein preferably said asthma is allergic asthma, and wherein said method comprises (i) selecting a patient on controller treatment, wherein said patient is controlled by said patient's controller treatment, wherein said patient's controller treatment is treatment by way of inhaled glucocorticosteriods or by way of inhaled glucocorticosteriods combined with long-acting p₂-agonists, preferably by way of inhaled glucocorticosteriods, and wherein said patient has an eosinophilic cell count of greater than 0.1 cells/nl in peripheral blood, preferably equal to or greater than 0.15 cells/nl; and (ii) administering an effective amount of said composition to said patient, wherein said composition is an immune-modulating substance, wherein said immune-modulating substance is packaged in a virus-like particle of a R A bacteriophage Q consisting of coat proteins consisting of the amino acid sequence of SEQ ID NO:7, and wherein said immune-modulating substance consists of an immune-modulating nucleic acid, and wherein said immune-modulating nucleic acid is an unmethylated CpG containing oligonucleotide consisting of the sequence GGGGGGGGGGGACGATCGTCGGGGGGGGGG (SEQ ID NO: 6), and wherein said unmethylated CpG-containing oligonucleotide consists exclusively of phosphodiester bound nucleotides; and wherein said administration is effective to improve asthma control of said patient.

In a further very preferred embodiment, the present invention provides a composition for use in a method for treating asthma of a patient in need thereof, and wherein preferably said asthma is allergic asthma, and wherein said method comprises (i) selecting a patient on controller treatment, wherein said patient is controlled by said patient's controller treatment, wherein said patient's controller treatment is treatment by way of inhaled glucocorticosteriods or by way of inhaled glucocorticosteriods combined with long-acting ₂-agonists, preferably by way of inhaled glucocorticosteriods, and wherein said patient has an eosinophilic cell count of greater than 0.1 cells/nl and less than 0.3 cells/nl in peripheral blood preferably of greater than 0.15 cells/nl and less than 0.25 cells/nl; and (ii) administering an effective amount of said composition to said patient, wherein said composition is an immune-modulating substance, wherein said immune -modulating substance is packaged in a virus-like particle of a RNA bacteriophage Q consisting of coat proteins consisting of the amino acid sequence of SEQ ID NO:7, and wherein said immune-modulating substance consists of an immune-modulating nucleic acid, and wherein said immune-modulating nucleic acid is an unmethylated CpG containing oligonucleotide consisting of the sequence GGGGGGGGGGGACGATCGTCGGGGGGGGGG (SEQ ID NO:6), and wherein said unmethylated CpG-containing oligonucleotide consists exclusively of phosphodiester bound nucleotides; and wherein said administration is effective to improve asthma control of said patient.

In further preferred embodiments to the aforementioned very preferred ones, said patient's controller treatment is treatment by way of low dose inhaled glucocorticosteriods, or said patient's controller treatment is treatment by way of medium dose inhaled glucocorticosteriods or said patient's controller treatment is treatment by way of high dose inhaled glucocorticosteriods.

In further preferred embodiments to the aforementioned very preferred ones, said patient's controller treatment is treatment by way of low dose inhaled glucocorticosteriods combined with long-acting p₂-agonists, or said patient's controller treatment is treatment by way of medium dose inhaled glucocorticosteriods combined with long-acting ₂-agonists, or said patient's controller treatment is treatment by way of high dose inhaled glucocorticosteriods combined with long-acting ₂-agonists.

Further preferred embodiments to the aforementioned very preferred ones have been described throughout this description.

EXAMPLES

Example 1

Treatment of patients with mild-to-moderate persistent allergic asthma with QbGlO The clinical trial was conducted as described in Beeh (Beeh, K.-M., et al., J Allergy Clin

Immunol 131 :866-874 (2013)). Briefly, 63 patients that were controlled on ICSs alone prior to the treatment phase with QbGlO as indicated by an ACQ-7 score of <1.5 and whose allergic asthma symptoms required long-term treatment with ICS doses of >50(^g beclomethasone dipropionate (BDP) equivalent (corresponding thus to mild-to-moderate persistent asthma in accordance with GINA - Global Initiative for Asthma; Global Strategy for Asthma Management and Prevention (2012 update); available at http://www.ginasthma.org) were enrolled in this double-blind, parallel-group trial at five sites. Randomized patients were allocated to receive either seven injections 0.9 mg QbGlO (n=33) or placebo (n=30), respectively. Prior to randomization patients were converted to a run-in daily dosage of either 400 μg (for patients on 500-800μg BDP equivalent) or 800μg BDP (for patients on > 800μg BDP equivalent) inhaled morning and evening. The first three subcutanous injections of study drug were given weekly and then every second week on opposite upper arms, alternatively. During the first four weeks, patients were maintained on 100% BDP. Then the dose was reduced to 50% for the following four weeks. At the end of week eight BDP was completely withdrawn for the last four weeks, if clinically justifiable and following pre-defined safety criteria.

Throughout the trial, patients completed daily an asthma diary on special paper (dotforms^®, PharmaForms GmbH, Germany) and with a digital pen with an integrated EDC (Electronic Data Capture) solution (AMEDON GmbH, Germany) facilitating compliance monitoring and ensuring adherence to its completion. The following items were documented: day- and nighttime asthma symptoms, morning peak expiratory flow (PEF) measured with a portable peak flowmeter, puffs of relief medication salbutamol per 24 hours, and dosage of morning/evening inhalation of BDP (Cyclocaps^® Beclomethasone-dipropionate, single dose-dry-powder inhaler, PB Pharma, 100μg, 200μg, or 400μg capsules).

The average symptom and medication score (ASMS; score 0-7) calculated as the mean value of average asthma symptom score (ASS, score 0-8) and average asthma medication score (AMS, score 0-6) was determined throughout the trial. ASS was defined as the sum of the day- and nighttime asthma scores, each with a score range of 0 (no symptoms) to 4 (asthma very bad; bad night). AMS (score 0-6) was based on the number of puffs of salbutamol per 24 hours, using the same scoring as used in the 7-item asthma control questionnaire (ACQ-7; (Juniper, E. F., et al. Eur Respir J 14:32-38 (1999)). The ACQ-7 (score 0-6) represents the mean of responses to seven items including frequency and severity of daytime and nighttime symptoms, limitations of activities within the preceding seven days, a measure for salbutamol use, and measured FEVi. This validated questionnaire was indispensable due to safety reasons during the steroid reduction phase. Patients were subjected to next lower BDP levels only upon predefined safety criteria on asthma control and the investigator's clinical judgment. Furthermore, pulmonary function was measured by means of spirometry according to ATS/ERS guidelines. Additional exploratory outcomes were fraction of exhaled nitric oxide (FeNO), measurement of blood eosinophils, and airway- hyperresponsiveness to methacholine challenge recorded as PC20. The analysis of said blood eosinophils, and hereby the analysis of the eosinophilic cell count in peripheral blood was conducted by the automated analyzer Roche Sysmex XE-2100 using a laser based flow cytometer as analytical module. The total number of eosinophil cells in peripheral blood was provided in counts/nl (10^~9 liter), wherein the number was rounded to the first figure after the decimal. The rounding was as follows: if the second figure after the decimal was a "0", "1", "2", "3" or "4", then it was down rounded, wherein if the second figure after the decimal was a "5", "6", "7", "8" or "9", then it was up rounded. For example, a value of 0.15 was rounded up to 0.2, wherein a value of 0.24 was rounded down to 0.2.

The data from all randomized subjects were analyzed on an intention-to-treat (ITT) basis. For ITT analyses, all randomized subjects were included who were exposed to study drug and for whom at least one post-baseline documentation of efficacy was available. For all efficacy parameter last observations were carried forward (LOCF) for drop outs. In case of deviations to the planned steroid reduction scheme, values of the last visit with steroid dose on plan were carried forward as well. Example 2

Patients with an eosinophil cell count in peripheral blood of more than 0.1 cells/nl benefit from their controller treatment

The asthma patients treated as described in Example 1 were stratified according to eosinophil cell counts in peripheral blood at baseline on controller treatment. The stratification of the patients was, on one hand, patients having an eosinophil cell count in peripheral blood of equal to or less than 0.1 cells/nl, and on the other hand, patients having an eosinophil cell count in peripheral blood of greater than 0.1 cells/nl.

The change in ACQ-7, FEVi and exhaled nitric oxide (FeNO) was analysed in placebo treated patients whilst controller ICS treatment was gradually reduced from 100 % to 0 %. Briefly, controller ICS treatment was maintained on 100 % during the first four weeks and than reduced to 50%. After further 4 weeks, i.e. at week 8, the controller ICS treatement was completely removed for the last 4 weeks of the treatment. Change from baseline after 4, 8 and 12 weeks of treatment was determined for placebo treated patients in both experimental placebo patient groups, i.e. on one hand, placebo patients having an eosinophil cell count in peripheral blood of equal to or less than 0.1 cells/nl, and on the other hand, placebo patients having an eosinophil cell count in peripheral blood of greater than 0.1 cells/nl.

As shown in Table 1, placebo patients which had an eosinophil cell count in peripheral blood of greater than 0.1 cells/nl.at baseline deteriorated much more upon controller treatment withdrawal over time than the ones that had an eosinophil cell count in peripheral blood of equal to or less than 0.1 cells/nl. Note that deterioration of asthma control is characterized by a positive value for AACQ-7 and AFeNO, and a negative value for AFEVi. Taken together these results demonstrate that controller treatment is much more effective in patients with an eosinophil cell count in peripheral blood of greater than 0.1 cells/nl as compared to the ones having an eosinophil cell count in peripheral blood of equal to or less than 0.1 cells/nl. Hence, only patients with an eosinophil cell count in peripheral blood of greater than 0.1 cells/nl clearly benefit from controller treatment.

Tabel 1 : Change of ACQ-7, FeNO and FEVi from baseline upon controller treatment reduction in placebo patients with < 0.1 Eosinophils/nl in peripheral blood versus placebo patients with >0.1 Eosinophils/nl in peripheral blood.

Example 3

Patients with an eosinophil cell count in peripheral blood of greater than 0.1 cells/nl benefit from treatment with QbGlO on top of 100 % controller treatment

The asthma patients treated as described in Example 1 were stratified according to peripheral blood Eosinophil counts at baseline on controller treatment. The stratification of the patients was, on one hand, patients having an eosinophil cell count in peripheral blood of equal to or less than 0.1 cells/nl, and on the other hand, patients having an eosinophil cell count in peripheral blood of greater than 0.1 cells/nl.

The change in ACQ-7, FEVi and exhaled nitric oxide (FeNO) was compared between patients treated with QbGlO and patients treated with placebo. Briefly, change from baseline after 4 weeks of treatement was determined for patients treated with QbGlO and patients treated with placebo. To evaluate the QbGlO treatment effect the change from baseline achieved with placebo after 4 weeks was deducted from the one obtained in patients treated with QbGlO after 4 weeks in both experimental, i.e. on one hand, patients having an eosinophil cell count in peripheral blood of equal to or less than 0.1 cells/nl, and on the other hand, patients having an eosinophil cell count in peripheral blood of greater than 0.1 cells/nl.

As shown in Table 2, a much better improvement was observed on top of controller treatment in the patients which had an eosinophil cell count in peripheral blood of greater than 0.1 cells/nl than in the ones that had an eosinophil cell count in peripheral blood of equal to or less than 0.1 cells/nl. The ACQ-7 was improved by 0.49 units in the group > 0.1 Eosinophils per nl peripheral blood, whilst it deteriorated by 0.04 units in patients with < 0.1 Eosinophils per nl peripheral blood. Likewise the lung function improved by 127 ml in patients with > 0.1 Eosinophils per nl peripheral blood whilst it deteriorated by 37 ml in patients with < 0.1 Eosinophils per nl peripheral blood. Last but not least, FeNO was improved by -6.46 ppb in the group > 0.1 Eosinophils per nl peripheral blood, whilst it deteriorated by 0.56 ppb in patients with < 0.1 Eosinophils per nl peripheral blood. It is important to note, that patients were maintained on a full standardized controller treatment with ICS based on the patients needs. Taken together these results demonstrate that treatment with QbGlO was effective on top of controller treatment in patients with > 0.1 Eosinophils per nl peripheral blood.

Table 2: ΔΔ ACQ-7, FEVi and FeNO (Change from baseline QbGlO - Change from baseline Placebo) after 4 weeks of treatment

Example 4

Patients with an eosinophil cell count in peripheral blood of more than 100 cells/μΐ benefit from treatment with QbGlO even if controller treatment was reduced by 50%

The asthma patients treated as described in Example 1 were stratified according to peripheral blood Eosinophil counts at baseline on controller treatment. The stratification of the patients was, on one hand, patients having an eosinophil cell count in peripheral blood of equal to or less than 0.1 cells/nl, and on the other hand, patients having an eosinophil cell count in peripheral blood of greater than 0.1 cells/nl.

The change in ACQ-7, FEVi and exhaled nitric oxide (FeNO) was compared between QbGlO treated and placebo treated patients. Briefly, change from baseline after 8 weeks of treatment was determined for QbGlO treated and for placebo treated patients. At this time point the controller treatement consisting of ICS had been reduced by 50 % as compared to the controller ICS treatement at baseline. To evaluate the QbGlO treatment effect in this patient population treated with reduced amounts of ICS, the change from baseline achieved with placebo after 8 weeks was deducted from the one obtained in patients treated with QbGlO after 8 weeks in both experimental groups (< 0.1 Eosinophils per nl peripheral blood versus >0.1 Eosinophils per nl peripheral blood).

As shown in Table 3, a much better improvement was observed on top of reduced controller treatment in the patients which had an eosinophil count >0.1 Eosinohils per nl peripheral blood than in the ones that had an eosinophil count < 0.1 Eosinophils per nl peripheral blood. The ACQ-7 was improved by 0.93 units in the group > 0.1 Eosinophils per nl peripheral blood, whilst only an improvement of 0.1 units was observed in patients with < 0.1 Eosinophils per nl peripheral blood. Likewise the lung function improved by 381 ml in patients with > 0.1 Eosinophils per nl peripheral blood whilst it only improved by 98 ml in patients with < 0.1 Eosinophils per nl peripheral blood. Lastly FeNO was improved by 12.35 ppb in the group > 0.1 Eosinophils per nl peripheral blood, whilst it was only reduced by 1.07 ppb in patients with < 0.1 Eosinophils per nl peripheral blood. Taken together these results demonstrate that treatment with QbGlO was effective and even led to an improvement although controller treatment was reduced by 50% in patients with > 0.1 Eosinophils per nl peripheral blood.

Table 3: ΔΔ ACQ-7, FEVi and FeNO (Change from baseline QbGlO - Change from baseline Placebo) after 8 weeks of treatment

AAACQ-7 AAFEVi (ml) AAFeNO (ppb) (QbGlO -Placebo) (QbGlO -Placebo) (QbGlO-Placebo)

< 0.1 Eosinophils per nl

- 0.1 98 -1.07 peripheral blood

> 0.1 Eosinophils per nl

- 0. 93 381 -12.35 peripheral blood Example 5

Patients with an eosinophil cell count in peripheral blood of greater than 0.1 cells/nl benefit from treatment with QbGlO even in the absence of controller treatment

The asthma patients treated as described in Example 1 were stratified according to peripheral blood Eosinophil counts at baseline on controller treatment. The stratification of the patients was, on one hand, patients having an eosinophil cell count in peripheral blood of equal or less than 0.1 cells/nl, and on the other hand, patients having an eosinophil cell count in peripheral blood of more than 0.1 cells/nl.

The change in ACQ-7, FEVi and exhaled nitric oxide (FeNO) was compared between QbGlO treated and placebo treated patients. Briefly, change from baseline after 12 weeks of treatement was determined for QbGlO treated and for placebo treated patients. At this time point the controller treatment consisting of ICS had been completely removed. To evaluate the QbGlO treatment effect in this patient population treated with reduced amounts of ICS, the change from baseline achieved with placebo after 12 weeks was deducted from the one obtained in patients treated with QbGlO after 12 weeks in both experimental groups (< 0.1 Eosinophils per nl peripheral blood versus >0.1 Eosinophils per nl peripheral blood).

As shown in Table 4, a much better improvement was observed on top of removed controller treatment in the patients which had an eosinophil count >0.1 Eosinohils per nl peripheral blood than in the ones that had an eosinophil count < 0.1 Eosinophils per nl peripheral blood. The ACQ-7 was improved by 1.2 units in the group > 0.1 Eosinophils per nl peripheral blood, whilst no improvement was observed in patients with < 0.1 Eosinophils per nl peripheral blood. Likewise the lung function improved by 402 ml in patients with > 0.1 Eosinophils per nl peripheral blood whilst it only improved by 21 ml in patients with < 0.1 Eosinophils per nl peripheral blood. Taken together these results demonstrate that treatment with QbGlO was effective and even led to an improvement although controller treatment was completely removed in patients with > 0.1 Eosinophils per nl peripheral blood. Lastly FeNO was improved by 16.96 ppb in the group > 0.1 Eosinophils per nl peripheral blood, whilst it deteriorated by 8.25 ppb in patients with < 0.1 Eosinophils per nl peripheral blood.

Table 4: ΔΔ ACQ-7, FEVi and FeNO (Change from baseline QbGlO - Change from baseline Placebo) after 12 weeks of treatment

AAACQ-7 AAFEVi (ml) AAFeNO (ppb) (QbGlO-Placebo) (QbGlO-Placebo) (QbGlO-Placebo)

< 0.1 Eosinophils per nl

0 21 8.25 peripheral blood > 0.1 Eosinophils per nl

-1.17 402 -16,96 peripheral blood

Example 6

Stratification of ACQ-7 based on peripheral blood eosinophil counts (improvement of QbGlO compared to Placebo)

The asthma patients treated as described in Example 1 were stratified according to peripheral blood Eosinophil counts at baseline on controller treatment. The stratification of the patients was based on eosinophil cell count in peripheral blood on controller treatment. The following groups were made: 0.0 cells/nl, 0.1 cells/nl, 0.2 cells/nl, 0.3 cells/nl and 0.4 cells/nl. The change in ACQ-7 was compared between QbGlO treated and placebo treated patients. Briefly, change from baseline after 4, 8 and 12 weeks of treatement was determined for QbGlO treated and for placebo treated patients. To evaluate the QbGlO treatment effect in this patient population treated with reduced amounts of ICS, the change from baseline achieved with placebo after 4, 8 and 12 weeks was deducted from the one obtained in patients treated with QbGlO after 4, 8 and 12 weeks in all five experimental groups and averages over the 4 time points was made for all five experimental groups. As shown in Table 5, a much better improvement was observed on top of reduced controller treatment in the patients which had an Eosinophil count > 0.1 cells than in the ones that had an Eosinophil count < 0.1. Importantly all investigated groups with an Eosinohil count >0.1 cells benefited from the treatment. These results demonstrate that treatment with QbGlO was effective in patients with an Eosinophil count >0.1 cells and at least up to 0.4 cells/nl.

Table 5: ΔΔ ACQ-7 (Change from baseline QbGlO - Change from baseline Placebo) after 4, 8 and 12 weeks weeks of treatment.

Claims

Claims

A composition for use in a method for treating asthma of a patient in need thereof, wherein said method comprises

(i) selecting a patient on controller treatment, wherein said patient is controlled by said patient's controller treatment, and wherein said patient has an eosinophilic cell count of greater than 0.1 cells/nl in peripheral blood; and

(ii) administering an effective amount of said composition to said patient, wherein said composition comprises, preferably consists of,

(a) an immune -modulating substance;

(b) a long-acting p₂-agonist;

(c) an inhaled glucocorticosteriod; or

(d) any mixture of at least one of (a), (b) and/or (c)

wherein said administration is effective to improve asthma control of said patient.

The composition of claim 1 , wherein said patient has an eosinophilic cell count of less than 0.3 cells/nl in peripheral blood.

The composition of any one of the preceding claims, wherein said patient's controller treatment is treatment by way of inhaled glucocorticosteriods, wherein preferably said patient's controller treatment is treatment by way of low, medium or high dose inhaled glucocorticosteriods, or wherein said patient's controller treatment is treatment by way of inhaled corticosteroids combined with long-acting ₂-agonists, wherein preferably said patient's controller treatment is treatment by way of low, medium or high dose inhaled glucocorticosteriods combined with long-acting ₂-agonists.

The composition of any one of the preceding claims, wherein said patient's controller treatment is continued, reduced or removed during said administering said effective amount of said composition to said patient.

The composition of any one of the preceding claims, wherein said composition comprises, preferably consists of, an immune -modulating substance, wherein preferably said immune- modulating substance comprises, preferably consists of,

(i) an immune-modulating nucleic acid; (ii) a toll-like receptor 7 ligand (TLR7-ligand);

(iii) a toll-like receptor 9 (TLR9-ligand);

(iv) a chemoattractant receptor-homologous molecule expressed on Th2

cells antagonist (CRTh2 anatagonist);

(v) a chemokine antagonist;

(vi) a somatotaxin; or

(vii) any mixture of at least one of (i), (ii), (iii), (iv), (v) and/or (vi);

and wherein preferably said composition comprises, preferably consists of, an immune- modulating substance, wherein said immune-modulating substance comprises, preferably consists of:

(i) an immune-modulating nucleic acid;

(ii) a TLR7-ligand;

(iii) a TLR9-ligand; or

(iv) any mixture of at least one of (i), (ii) and/or (iii).

The composition of any one of the preceding claims, wherein said composition comprises, preferably consists of, an immune-modulating substance, wherein said immune -modulating substance comprises, preferably consists of, an immune-modulating nucleic acid, wherein preferably said immune-modulating nucleic acid is capable of stimulating IFN-alpha production in a cell.

The composition of any one of the preceding claims, wherein said composition comprises, preferably consists of, an immune-modulating substance, wherein said immune -modulating substance comprises, preferably consists of, a TLR7-ligand, or wherein said composition comprises, preferably consists of, an immune-modulating substance, wherein said immune- modulating substance comprises, preferably consists of, a TLR9-ligand; and wherein further preferably said composition comprises, preferably consists of, an immune- modulating substance, wherein said immune-modulating substance comprises, preferably consists of, a TLR9-ligand.

The composition of any one of claims 1 to 7, wherein said composition comprises, preferably consists of, an immune-modulating substance, wherein said immune -modulating substance comprises, preferably consists of an unmethylated CpG containing oligonucleotide, wherein preferably said unmethylated CpG containing oligonucleotide is selected from of: (a) A-type CpGs;

(b) B-type CpGs; or

(c) C-type CpGs;

and wherein further preferably said unmethylated CpG-containing oligonucleotide is an A- type CpG.

9. The composition of claim 8, wherein said unmethylated CpG-containing oligonucleotide comprises, or alternatively consists essentially of, or alternatively consists of the sequence GGGGGGGGGGGACGATCGTCGGGGGGGGGG (SEQ ID NO:6), and wherein preferably said unmethylated CpG-containing oligonucleotide consists exclusively of phosphodiester bound nucleotides.

The composition of any one of the preceding claims, wherein said immune-modulating nucleic acid is an unmethylated CpG containing oligonucleotide consisting of the sequence GGGGGGGGGGGACGATCGTCGGGGGGGGGG (SEQ ID NO: 6), and wherein said unmethylated CpG-containing oligonucleotide consists exclusively of phosphodiester bound nucleotides.

11. The composition of any one of the preceding claims, wherein said composition comprises, preferably consists of, an immune-modulating substance, wherein said immune -modulating substance comprises, preferably consists of:

(i) an immune-modulating nucleic acid;

(ii) a TLR7-ligand;

(iii) a TLR9-ligand; or

(iv) any mixture of at least one of (i), (ii) and/or (iii);

wherein said immune-modulating substance is packaged in a particle, wherein said particle is, preferably, a virus-like particle or a virosome.

12. The composition of claim 1 1, wherein said particle is a virus-like particle, wherein preferably said virus-like particle is a virus-like particle of a RNA bacteriophage Q , and wherein further preferably said virus-like particle of a RNA bacteriophage Q consists of coat proteins having the amino acid sequence of SEQ ID NO:7.

13. The composition of any one of the preceding claims, wherein said method does not comprise co-administering an allergen to said patient, and wherein said composition does not comprise an allergen, and wherein preferably said composition is not administered to said patient in conjunction with an allergen.

14. The composition of any one of the preceding claims, wherein said asthma is allergic asthma.

15. The composition of any one of the preceding claims, wherein said administration is effective to improve asthma control of said patient as measured by either a reduction of ACQ-7 of at least of 0.3, preferably 0.5 points.

16. A pharmaceutical composition for use in a method for treating asthma of a patient in need thereof, said pharmaceutical composition comprising, essentially consisting of, or consisting of:

(a) a composition of any one of claims 1 to 15; and

(b) a pharmaceutically acceptable carrier.