GB2379875A

GB2379875A - Immunity generation

Info

Publication number: GB2379875A
Application number: GB0217035A
Authority: GB
Inventors: Christopher Jeremy Leonard
Original assignee: Individual
Current assignee: Individual
Priority date: 2001-07-23
Filing date: 2002-07-23
Publication date: 2003-03-26
Also published as: GB0217035D0; GB0117755D0

Abstract

A method for the manufacture of a medicament for immunity generation includes providing an anti-bacterial peptide, and causing the anti-bacterial peptide to react with a bacterium, virus, fungus, parasite or mycobacterium to produce a reaction product which can be delivered as an antigen complex via the chemotactic T-cell pathway to promote an adaptive immune system. The antibacterial peptide(s) are preferably sourced from insect tissues and/or larval forms.

Description

IMMUNITY GENERATION Field of the Invention This invention relates to immunity generation.

In PCT Specification No. WO 00/74693, to which reference should be made, there is described a method for the manufacture of a medicament for immunity generation, which method includes the use of insect tissues and/or larval forms and derivatives of insects.

It is an object of the present invention to provide an improved method for the manufacture of a medicament for immunity generation.

Further objects of the present invention include the provision of a method for the manufacture of a probiotic food for animals and

humans, and a method for the manufacture of anti-inflammatory compounds, specifically compounds for preventing or reducing inflammation of the gut and airway surfaces.

Summary of the Invention According to a first aspect of the present invention there is provided a method for the manufacture of a medicament for immunity generation, which method includes providing an antibacterial peptide, and causing the anti-bacterial peptide to react with a bacterium, virus, fungus, parasite or mycobacterium to produce a reaction product which can be delivered as an antigen complex via the chemotactic T-cell pathway to promote an adaptive immune system.

More than one anti-bacteriai peptide may be provided and may be caused to react with more than one bacterium, virus, fungus, parasite and/or mycobacterium to produce a complex reaction product in vitro.

The complex reaction product can then be delivered as an antigen complex, i. e. as a vaccination/immunisation strategy, via ingestion to the gut or nasally to the airway surface or subcutaneously by injection.

The reaction product (antigen) then acts, via the chemotactic T-cell/immature dendrite pathway and through the regulatory effects on co-stimulatory molecule expression by lymphocytes, to promote an adaptive immune response.

The anti-bacterial peptide (s) is or are preferably prepared from insect tissues and/or larval forms and/or derivatives and

isolated using generally available separation techniques. Reference should be made to PCT Specification No. WO 00/74693 for further information in this respect.

The anti-bacterial peptide (s) will react selectively within the insect larvae to kill or inactivate pathogenic bacteria but not to kill non-pathogenic probiotic bacteria. In this way, the reaction product can be used as a food for animals or humans and becomes a noncarrier of pathogenic bacteria but a provider of probiotic bacteria which, on ingestion, help to promote the ingestor's intestinal probiotic bacterial fauna content.

Thus, according to a second aspect of the present invention there is provided a method for the manufacture of a medicament for immunity generation, which method includes providing an antibacterial peptide, and causing the anti-bacterial peptide to react with a bacterium, virus, fungus, parasite or mycobacterium to produce a reaction product.

In providing a boost to the innate immune system to enable it to respond to bacterial challenge, the reaction product also acts as an anti-inflammatory agent for the gut and airway surfaces by decreasing the need for excessive adaptive immune response.

Thus, according to a third aspect of the present invention there is provided a method for the manufacture of a compound for preventing or reducing inflammation of the gut and airway surface, which method includes providing an anti-bacterial peptide, and

causing the anti-bacterial peptide to react with a bacterium, virus, fungus, parasite or mycobacterium to produce a reaction product which can be delivered as an antigen complex via the chemotactic T-cell pathway to promote an adaptive immune system.

The method of preparation from insect tissues and/or larval forms and/or derivatives of insects may be such as to produce a plurality of anti-bacterial peptides which produce antigens which have a positive effect on the acquired immune mechanism of the recipient.

The insect tissues and/or larval forms and/or derivatives of insects may be any of those described in PCT Specification No.

WO 00/74693. They may also be any one or more of the following :- 1) large water beetle-hydrophilidae, 2) blister beetle-meloidae, 3) dung beetle, 4) may beetle-cockchafer, 5) termites-isoptera, 6) moths-saturnids, 7) grasshoppers/crickets-orthoptera, 8) crickets-acheta domestica, 9) wichetty grubs-cossidae, 10) wood lice-isopoda, and 11) cochineal-dactylopius coccus.

Description of the Preferred Embodiments The provision and direct ingestion of anti-microbial peptides by an ingestor provides it with an enhanced platform of defence molecules to resist a first wave of pathogenic challenge, taking the pressure off the adaptive immune response system of the ingestor.

Inflammatory response occurs both in the gut airway and throughout the body for, for example, rheumatoid arthritis, atherosclerosis, Parkinson's disease, Alzeimer's disease and Crohn's disease.

Current methodologies for the treatment of these diseases are centred on the use of drugs which control various aspects of the adaptive response system. In contrast therewith, anti-microbial peptides act in a feed-back loop suppressing the activation of the adaptive immune response system and thus restricting the damage caused by self-inflicted adaptive response attack. The antimicrobial peptides can be obtained either from ingested insects or through the absorption of the peptides produced by probiotic bacteria. Probiotic bacteria are a source of D-amino acids, which can be utilised by a host animal in the production of more effective isomers of anti-microbial agents, as opposed to the all L forms of the amino acids produced by the host animal.

The anti-microbial peptides may be used by the host animal as surveillance molecules for cancer and tumour cells, promoting their antigenecity and making them more susceptible to T cell

lymphocytes and the production of host-created anti-bodies (including memory cells and the necessary pathway precursors and products) that maintain an effective on-going screening process within the host animal, unlike current products which involve antibody production without memory cells.

It has been found that many of the gut probiotics cannot be grown in vitro and the best and most efficient manner of promoting and making available such bacteria is outlined below.

In addition to the species outlined in PCT Specification No.

WO 00/74693, the following insects may also be used : - 1) large water beetle-hydrophilidae, 2) blister beetle-meloidae, 3) dung beetle, 4) may beetle-cockchafer, 5) termites-isoptera, 6) moths-saturnids, 7) grasshoppers/crickets-orthoptera, 8) crickets-acheta domestica, 9) wichetty grubs-cossidae, 10) wood lice-isopoda, 11) cochineal-dactylopius coccus, 12) locusts, and 13) sap-consuming Homoptera aphids.

The ingestion of such insectsas food by animals, e. g. chickens, pigs, man, etc. confers on the ingestor a compatible

digesting bacterial fauna corresponding to that which was successfully operating in the insects.

In this way, selected probiotic bacteria are used to fill the gut, as opposed to the possible array of unsuitable airborne pathogenic bacteria, or exposure to and ingestion of soil pathogenic bacteria, causing health challenges to the young.

Current farming methodologies do not take into account the natural process of specific bacterial fauna formulation in the developments of chickens, pigs, cows and the like. Much work has been done on the promotion of lacto bacillus, but it has now been shown that, for adults, this represents only 2% of gut fauna, possibly representing the equivalent position of milk in the diet of adults. Other probiotic bacteria are necessary that represent the natural species which are capable of breaking down complex sugars and other molecules, using enzymes and metabolic pathways which are not otherwise available to the host animal.

The bacterial fauna of an insect's gut is normally provided by the presentation by the female adult of a suitable assortment of gut bacteria, either as a package, as in the case of the stink bug, or through f cal pellets left on the food substrate on which the eggs have been laid, as is the case of the blowfly. This replicates the lacto bacillus presentation in human milk and colostrums to neonates.

Likewise, other insects, such as ants, bees, wasps and various beetles, provide regurgitated food that will be mixed with the adults'gut bacterial fauna. In this way, bacteria that have developed successfully in the adult insects are transferred to the young, where theses advantageous biochemical syntheses can be enjoyed by the next generation of insects, providing sources of vitamins and essential amino acids and, as in the case of termites, biochemical pathways capable of digesting cellulose and presenting usable bi-products for utilisation by the insects.

Bacterial inactivation may occur within a liquified growth medium, to which the peptides are added in a suitable homeostatic medium. Introduced nasally, the reaction product may represent the equivalent of defensin killed bacteria, which are usually flushed from the airways in a mucus envelope and re-presented to the gut epithelia, at which point the adaptive immune response system can become involved. The adaptive immune response system may also be involved in the airway surface epithelia on nasal inhalation using proprietary systems for presentation. When given orally, the reaction product may be mucus-enveloped or chitosamencapsulated.

The anti-microbial peptides may be sourced from insects or created synthetically representing the broad spectrum of insectanimal-plant anti-microbial peptide array. These may also include anti-microbial peptides produced by gut bacteria themselves. These are produced to maintain the species equilibria of gut bacterial fauna as expressed by the probiotic gut bacteria and whose action

can suppress the multiplication of ingested or gut-presented pathogenic bacteria.

Many vaccination strategies have incorporated either live or dead bacteria or viruses. The live bacteria or viruses are generally more reactive and positive in producing an adaptive response but, in certain circumstances, they may be too active and result in development of the disease itself. The dead bacteria which are employed in such vaccination process are usually killed by physical process, for example, by heat or by chemical inactivation.

In contrast to the known vaccination strategies, the method of disease prevention contemplated by the present invention is more natural and takes account of the processes that develop during immune response. It thus takes account not only of the development of the challenge in the individual but also the evolutionary development from the simple innate response in insects to the more developed aspects of adaptive immunity in higher animals. The adaptive response may also develop in the presence of peptide-killed bacteria exposed sections for creating new antigen modes through new chemical affinities. Many of the bacteriocins are produced as inactive precursors and their activation occurs on cleavage by various proteases.

Natural growth factors that are associated with the ingestion of colostrums in mammals can be provided by insects in poultry development. It is known that many beneficial peptides act synergenically as is provided by ingestion of a whole insect diet.

The peptides are effectively coated in a resistant chitin coat enabling the peptides to transfer down the gut before they are subjected to proteolytic attack.

Anti-microbial peptides can act in a feed-back loop to suppress the activity of tumour necrosis factor (TNF), which is overproduced in inflammatory bowel disease and rheumatoid arthritis.

Anti-microbial peptides are chaperoned and protected by a-2- macroglobulin, which is also a major serum protease inhibitor. The anti-microbial peptides may be selectively absorbed and chaperoned.

For the control of cystic fibrosis there is a requirement for anti-microbial peptides that are high in salt tolerance and the insects from which the peptides are derived can accordingly be selected to achieve this high salt tolerance. The selected peptides modify the inflammatory response through the regulation of cytokine production and adhesion molecule expression. It is also to be noted that anti-microbial peptides are chemotactic to monocytes, dendritic cells and T cells, including memory T cells via the chemokine receptor CCR6.

The method of preparation of the anti-microbial peptides from insect tissues and/or larval forms and/or derivatives of insects may be such as to produce a plurality of anti-bacterial peptides which produce antigens which have a positive effect on the acquired immune mechanism of the recipient.

Claims

Claims :-

1. A method for the manufacture of a medicament for immunity generation, which method includes providing an antibacterial peptide, and causing the anti-bacterial peptide to react with a bacterium, virus, fungus, parasite or mycobacterium to produce a reaction product which can be delivered as an antigen complex via the chemotactic T-cell pathway to promote an adaptive immune system.

2. A method as claimed in Claim 1, in which more than one anti-bacterial peptide is provided and is caused to react with more than one bacterium, virus, fungus, parasite and/or mycobacterium to produce a complex reaction product in vitro.

3. A method as claimed in Claim 2, in which the complex reaction product is delivered as an antigen complex via ingestion to the gut or nasally to the airway surface or sub-cutaneously by injection.

4. A method as claimed in any one of the preceding claims in which the anti-bacterial peptide (s) is or are prepared from insect tissues and/or larval forms and/or derivatives and isolated using generally available separation techniques.

5. A method as claimed in Claim 2, in which the reaction product is used as a food for animals or humans and becomes a non-carrier of pathogenic bacteria but a provider of probiotic

bacteria which, on ingestion, help to promote the ingestor's intestinal probiotic bacterial fauna content.

6. A method for the manufacture of a medicament for immunity generation, which method includes providing an antibacterial peptide, and causing the anti-bacterial peptide to react with a bacterium, virus, fungus, parasite or mycobacterium to produce a reaction product.

7. A method as claimed in Claim 6, in which, in providing a boost to the innate immune system to enable it to respond to bacteria ! challenge, the reaction product aiso acts as an antiinflammatory agent for the gut and airway surfaces by decreasing the need for excessive adaptive immune response.

8. A method for the manufacture of a compound for preventing or reducing inflammation of the gut and airway surface, which method includes providing an anti-bacterial peptide, and causing the anti-bacterial peptide to react with a bacterium, virus, fungus, parasite or mycobacterium to produce a reaction product which can be delivered as an antigen complex via the chemotactic T-cell pathway to promote an adaptive immune system.

9. A method as claimed in Claim 8, in which the reaction product is produced from insect tissues and/or larval forms and/or derivatives of insects in such manner as to produce a plurality of anti-bacterial peptides which produce antigens which have a positive effect on the acquired immune mechanism of the recipient.

10. Any new any useful method hereinbefore described.

11. The product obtained by any one of the preceding claims.