EA018848B1 - Immunologically active substance, process for preparing same, bioprobe based thereon, method for diagnostics of health state using bioprobe, pharmaceutical composition and method of treatment using pharmaceutical composition - Google Patents

Abstract

The claimed group of inventions relates to bioengineering and can be used in different medicine branches for diagnostics of an individual's tissue state, treatment of said tissue diseases characterised by an uncontrolled tissue cell growth, requiring stop grow thereof by local lysis inside the organism provided obligatory control by treating with the same substance which can be a base for laboratory testing and pharmaceutical composition. The claimed technical solutions are based on the production of a novel class of substances having immune activity which are obtained from the tissue of an internal, transitory or transplanted organ, a neoplasm (tumor) in the organism of an individual, have specific activity in relation to antibodies/autoantibodies and are characterized by the presence of properties which make it possible to identify, using a laboratory procedure, the condition of the tissue of the internal organs in an individual under examination by means of identifying an antigen-antibody or autoantigen-autoantibody reaction occurring between the substance and a biological liquid taken from the individual under examination and which have partial organ and species synergy, which makes it possible to induce a response of the immune system of an organism of an individual on parenteral administration which results in local lysis of the tissue having organ and species affinity to the administered substance.

Description

The technical field to which the invention relates.

The claimed group of inventions relates to biotechnology, can be used in medicine, veterinary medicine, laboratory diagnostics, immunology, in particular for diagnosing the condition of an individual's tissue, early diagnosis of an individual's tissue diseases, and also for treating diseases of an individual's tissue that can be characterized by uncontrolled growth of tissue cells (including, tissue tumors of a malignant or benign nature, intact developing in the body).

The level of technology

Recently, studies of autoantibodies, their role as predictors of processes developing in the body, have been actively conducted. There are publications that examine the relationship between autoantibodies and cancer. Publications are known in which the risk of stroke is predicted by the presence of certain autoantibodies in patients with atherosclerosis, and predictors of thrombosis or miscarriage in patients with antiphospholipid syndrome are studied. Conducted research on the search for autoantibodies as markers of other diseases.

The prior art is known immunologically active substances and methods for their production from tissues and organs of mammals, as well as technical solutions related to the diagnosis and treatment of various diseases. Since autoantibodies are a mirror image of autoantigens that are characteristic of various organs and tissues, their number and totality reflect the overall picture. It is known that in healthy people the number of those or other autoantibodies is about the same. A prolonged increase or decrease in their number indicates problems in the relevant authorities. Thus, autoantibodies are precursors not only of autoimmune diseases, but also of many others, in particular cardiovascular diseases, diseases of the gastrointestinal tract, kidneys, liver, lungs, etc.

In particular, a pharmaceutical composition for the treatment of immune diseases is known, containing a proteinaceous or non-proteinaceous substance that has activity in modulating LUM mediated signaling and a pharmaceutically acceptable carrier (RF application no. 2001114516, IPC: AK61 / 395). The substance has activity in inhibiting the proliferation of AYUM-expressing cells or in inhibiting the production of cytokine A1YMexpressing cells. The cytokine is interferon, which is a cytokine produced by T111-type T cells. or interleukin 4, which is a cytokine produced by T112-type T cells. The protein substance can be selected from the group consisting of an antibody that binds to AlIM or its part; a polypeptide containing fully or partially extracellular region A1IM; a fusion polypeptide containing a fully or partially extracellular region of A1IM and a fully or partially constant region of an immunoglobulin heavy chain; and a polypeptide that binds to A1IM. The non-protein substance may be DNA, RNA, or a chemically synthesized compound. The substance can be used to treat or prevent osteoarthritis, hepatitis, to treat or prevent a graft versus host reaction and immune rejection that accompanies the graft versus host reaction or a tissue or organ transplant. The substance can also be used to prevent an immune response triggered by a foreign antigen or autoantigen.

However, the claimed pharmaceutical composition has a different focus in the treatment of immune diseases and belongs to another class of substances (substances that prevent the immune response of antigens or autoantigens, affecting signal transmission (signal transduction)), has other purposes of treatment and therapeutic effects, can not be both substance suitable for diagnosis and treatment, and also does not imply individualization of diagnosis and treatment, cannot use microbes of an individual’s tissue to obtain substances but.

There is a method of assessing the risk of development and predisposition to the development of the pathology associated with the presence of autoantibodies against yrsg (application for the invention of the Russian Federation No. 2006132064, IPC: Ο01Ν33/564). The method consists in determining the presence of high levels of autoantibodies against endothelial receptor MS / activated MS (EPCB) in a sample taken from an individual, and provides a quantitative assessment of autoantibodies against EPCP ίη νίίτο in the specified sample, which can be used as a serum or plasma sample. The method further includes comparing the levels of anti-EPCB autoantibodies determined in the sample with the norm. These anti-EPCB autoantibodies can be quantified using an immunoassay using a marker or using an EB18A assay that involves immobilizing a polypeptide containing the amino acid sequence of EPC'P or its fragment containing at least one epitope that can be recognized on a solid carrier. anti-EBV autoantibody; incubating the immobilized polypeptide with a sample that is supposed to contain anti-EPCB autoantibodies and that was taken from the indicated individual for a period of time sufficient to bind antibodies to the immobilized polypeptide and form complexes of the polypeptide-anti-EPCB autoantibody; removing the remaining sample not bound to the immobilized polypeptide; incubation of polypeptide-anti-EPCB autoantibody complexes with the second anti

- 1 018848 by an enzyme conjugated antibody, wherein the second antibody is able to bind to said anti-EPTI autoantibodies. As said polypeptide, a polypeptide can be used that contains the amino acid sequence of a full-length URTP, or the amino acid sequence of an URFR fragment, which includes at least one epitope that can be recognized by an anti-URTI antibody. As a polypeptide, a hybrid protein can be used, containing region A, consisting of a polypeptide containing the amino acid sequence of the EHRF or its fragment comprising at least one epitope that can be recognized by an anti-EHRF antibody; or a region B consisting of a polypeptide comprising an amino acid sequence used to isolate or purify said hybrid protein, and / or an amino acid sequence used to immobilize said hybrid protein on a solid carrier. The specified polypeptide can be a hybrid protein, the amino acid sequence of which is presented in ZEC GO N0: 3. Amino acid sequence can be selected from AGD tags; Tags; EABAS tags; 8) ger-tags; an epitope that can be recognized by an antibody; 8BP tags; 3 tags; calmodulin binding peptide; cellulose binding domain; chitin binding domain; glutathione-3-transferase label; protein binding to maltose; ΝιΐδΑ. TghA, IKYA, Ανί-tags; A1a-Nsch-S1u-N18-Agg-Rgo (ZEZ GO N0: 4) (2, 4 and 8 copies), Rgo-Pe-Nsch-Akr-Nsch-AkrNsch-Rgo-Nk-Eei-Ua1-11e-Nsch -Leg (8EC) GO N0: 5), Sl-Me) -Tig-Cy8-X-X-Cy8 (W) GO N0: 6) (6 repeats), β galactosidase and V8U-glycoprotein. The specified second antibody specific for an immunoglobulin of a specific isotype is selected from an anti-human 1dS antibody, an anti-human 1dM antibody, an anti-human antibody | C | A, and mixtures thereof.

The presence of high levels of autoantibodies was associated with the pathology of the autoimmune disease, the system of lavish veterinarian, the ruling group, the vascular disease, and the vascular disease, as well as the vascular system, and the vascular system. blood circulation, limb ischemia, atherosclerosis, aneurysm, venous thrombosis and pulmonary embolism) and pregnancy complications (vykidy , Intrauterine fetal death, premature birth, delayed intrauterine growth, eclampsia and pre-eclampsia).

However, the claimed technical solution is used only as a diagnosis of certain nosological forms of the disease or of several diseases. Also, a known technical solution cannot be considered as a group of inventions representing a universal method and means used both for diagnostics and for treatment, for monitoring the course of treatment, for screening health by health profile, for identifying patients at risk of developing adverse reactions and prognosis. before mass vaccinations and (or) treatment with immunoactive substances. At the same time, it does not contain a scale for assessing the result, which allows to judge the stage or course of the diseases of the tissues studied. The claimed method may issue conclusions about the disease and / or diseases, but does not give a conclusion about the health status of the tissues of the organs of the individual and cannot directly assess the levels of their damage.

The considered technical solution is not capable of being strictly personalized and individualized, i.e. It is not applicable for a situation when an individual himself can be a source (donor) of tissue and this tissue can be used as a means of diagnosis and treatment in him (Individual X1 - donor = Individual X1 - patient). In addition, the solution in question cannot be used by individuals of the same species; in this case, any representative of the species can be a donor and other patients (Individual X1 - donor = Individual Xn - patient The individual, where X1 is one individual, and Xn - other individuals of the same species). In addition, the invention cannot be used for the treatment of cancer pathology.

Methods of immunoassay are also known (International Application for Invention \ ¥ 0 2006126008, application for the invention of the Russian Federation No. 2007149281, UK patent No. 2426581, IPC: S0Sh 33/564; S0Sh 33/574; S0Sh 33/564; C0Sh 33/574), according to which reveal a disease state or predisposition to the disease in mammals. The methods are based on detecting an antibody in a test sample containing a body fluid of said mammal, wherein said antibody is a biological marker of a disease state or susceptibility to the disease. The method includes contacting the specified test sample with a variety of different amounts of antigen specific for the indicated antibody, identifying the magnitude of specific binding of the indicated antibody and said antigen, plotting or calculating the curve of dependence of the magnitude of said specific binding on the amount of antigen for each antigen amount, and determining the presence or absence of a specified disease state or predisposition to the disease based on petsificheskogo binding between said antibody and said antigen for each antigen concentration used.

The antibody may be an autoantibody specific for a tumor marker protein, and the antigen may be a tumor marker protein or its antigenic fragment or epitope. The tumor marker protein is MIS1, MIS16, c-tus, ECNE, p53, hook, VASA1, VASA2, APC, NEJ2, P8A, CEA, CA19.9, CT-E80-1, 4-5, CACE, P8MA, Р8СА, ЕРСат, cytokeratin, recoverin, kallikrein, annexin,

- 2 018848

APP, CKP78, SA125, mammoglobin or gag. The method can be used in the diagnosis, prediction or monitoring of cancer, where a conclusion is made based on the comparison of the level of autoantibodies in the examined individuals with the values adopted as the norm. The method can also be applied in the selection of anticancer treatment for a particular patient patient, where the method is carried out using a panel of two or more antigens corresponding to different tumor marker proteins to determine the relative magnitude of the patient’s immune response to each of these different protein protein markers, where protein or tumor marker proteins that induce the strongest immune response or significant reactions in a patient are selected to develop the basis for an anti-cancer treatment of said the patient. Cancer treatment is a vaccination, and protein or tumor marker proteins that cause the strongest immune response or significant reactions in a patient are selected to develop the basis for an anticancer vaccine for a specified patient.

The antibody may be an autoantigen Addison, autoimmune hypoparathyroidism, autoimmune diabetes, or severe myasthenia) for kidney or liver disease leading to organ failure or failure, or related with such a disease, as well as for an autoimmune disease or associated with an autoimmune disease. An antigen is a natural protein or polypeptide, a recombinant protein or polynucleotide, a synthetic protein or polypeptide, a synthetic peptide, a peptidomimetic, a polysaccharide, or a nucleic acid.

However, in the above methods, the mechanism of immunoassay is presented and there is no information about the substance with which this mechanism can be carried out.

In the technical solution for the application for invention \ UO 2006126008, the substance participating in the laboratory reaction cannot simultaneously act as a pharmaceutical composition, when a) there are reasons requiring the response of the immune system of the animal organism; b) antibodies produced by the body must be antibodies not only against our substance, but must be autoantibodies against the tissue of the organism itself, leading to its local lysis. This solution does not use substances with partial syngenia, when one of the substances involved in the diagnosis is able to be a marker of its own antibodies and autoantibodies that occur when it is administered, and the tissues of the organ of the organism being examined, and with those and others supporting immune responses in the laboratory by engaging in parallel with specific antigen-antibody reactions. In addition, the description of this invention has a definition: An antibody can be an autoantibody specific for a tumor marker protein, and an antigen is a tumor marker protein or its antigenic fragment or epitope ... and further a tumor marker protein which can be is mis1, mis16, s-tus, ESKR, p53, hey, VKSA1, VKSA2, APC, NEK2, P8A, CEA, CA19.9,-Ε8Ο-1, 4-5, CACE, P8MA, P8CA, ErSat, cytokeratin , recoverin, kallikrein, annexation, APP, CKR78, CA125, mammo globin or GAH .... The disadvantage of the solution is the absence of substances with a partial syngene, and if the antibody (an autoantibody specific for a tumor marker protein, and a tumor marker protein or antigenic fragment or epitope) also has a partial (partially) syngeneic region (epitope) in its structure to the tumor tissue, it is test agent. Also the disadvantage of this method is the fact that the substance-antigen is not considered as a glycoprotein, in response to the introduction of which the body is able to produce antibodies belonging, at least, to the class of immunoglobulins M and C, as well as entering into immune reactions, at least with anti-M and anti-C immunoglobulins under laboratory conditions, having a partial (partial) syngenity to the tissue from which it is obtained, and devoid of sites responsible for the individual characteristics of the original tissue.

A method of obtaining substances from the tissues and organs of mammals with autoimmune activity ίη νίΐτο and ίη νίνο (RF patent for the invention No. 2240807, IPC: А61К 35/12, СШ 33/53). The method consists in the fact that the source material is typed, homogenized with quartz sand and 0.9% sodium chloride solution in a 1: 5 ratio, extracted on a Grasse with a 5-fold freezing cycle at -17 ° C and thawed at 37 ° C, centrifuged homogenate and isolate the supernatant, standardize the protein in it by the spectrometric method 20-22 mg / ml, freeze-dried. In addition, at the stage of the fourth freezing cycle, γ-quantum processing of the substance is carried out with 0.37 kJ γ-quanta, an integral dose of 1.8-104 g., For 20-25 minutes. Obtained by the claimed method, the substance retains the original organ specificity of the tissue and is deprived of the individual properties of the isoantigenic form of the source material of the donor. The combination of these properties provides greater accuracy of tissue identification in the reactions of η νίΐτο and shows its activity ίη νίνο.

The disadvantage of this method is its complexity in implementation, since it requires the use of special

- 3 018848 social equipment. The method uses radiation (gamma-quantum) technology, which requires special regulations for use, licensing, specific equipment, calibration and control devices. Compared with the known method, microwave technology is used, in which household ultrahigh frequencies are used, to influence the fabric, which makes the application of technology cheaper, more affordable and safer compared to radiation methods of exposure to the prototype fabric. In addition, in the claimed method of processing tissue, tissue disinfection operation is applied, which makes work with the initial substances safe, unlike the prototype method. The original tissue sample in the claimed method prior to the start of tissue typing and processing to obtain a substance can be preserved by freezing (up to -24 ° C) and stored for a long time at this temperature (up to 6 months) until it is used as a raw material for the production of without changing the quality of the substance, which also distinguishes the claimed technology from the prototype.

In addition, the substance obtained by a known method has an individual set of properties and structure that can be used exclusively in laboratory diagnostics and which cannot be extended to the treatment of tissue diseases. The substances of the prototype method are devoid of the properties of partial organ and species syngeney with the original organ properties retained. Substances obtained by the method according to the patent for the invention No. 2240807, have the original organ specificity of the tissue, which is devoid of the individual properties of the isoantigenic form of the source material of the donor tissue. The raw material of the fabric is subjected to various physical influences that cause various transformations in the original fabric. Radiation exposure to gamma rays has a cross-linking, ordering and strengthening rigid structure (intermolecular bonds) of the source material, and the microwave effect affects the hydrophilic bonds inside tissue biostructures, freeing free water in the tissue, causing the release of hydrophilic bonds inside the tissue and mobilizing free water molecules, leading to increasing tissue dehydration. The combination of these positions suggests that the substances obtained by the prototype method, and the claimed substances obtained by the proposed technology are fundamentally different. It was established that the substances of the above method can show their activity in the laboratory conditions of reactions of indirect hemagglutination (PHAA), which assess the state of autoimmunity of the organism. RNGA, being a routine method of immune responses, currently cannot be considered a standardizing method for immune studies. However, other methods of detecting specific autoantibodies, such as enzyme immunoassay, analysis of immune interactions of antigen-antibody (autoantigen-autoantibody) on solid carriers in the prototype method, have not been demonstrated, and the possibility of using them by other methods is not described. In addition, the disadvantage of this technical solution is the lack of the ability of the prototype substance to partially and isolated tissue lysis. The substances described in this patent are not able to act on the living tissue of the body without causing its death, and therefore cannot be used to compose pharmaceutical compositions, including for the treatment of cancer. They are not universal for the diagnosis and treatment (substance-substance can not be used for pharmaceutical and laboratory production).

DISCLOSURE OF INVENTION

The task of the group of inventions is to create a new biotechnology that can be used in medicine, veterinary medicine, laboratory diagnostics, immunology, pharmaceuticals for general and individual, personalized diagnostics of the condition of the tissue of an organ or tissues of an individual, treatment of diseases of these tissues that can be characterized by uncontrolled growth of tissue cells (including tissue tumors of a malignant or benign nature, intactly developing in the body) and requiring the cessation of such growth , by their local lysis inside a living organism in the implementation of mandatory control in the treatment of the same substance, which may be the basis for laboratory testing and pharmaceutical composition.

The technical result consists in the possibility of implementing a universal comprehensive direct diagnosis of the state of the tissue of one or more internal organs of the individual, while it is possible to promptly obtain a conclusion about the state of tissue health or the degree of damage in a living organism, which, in turn, makes it possible to judge the general state of health the body of the individual, personifying diagnosis and treatment. The diagnostic conclusion with the claimed method is formed faster, has a more accurate information and prognostic result, laboratory control data can detect pathological and morphological changes 24-72 hours before their clinical manifestation and the development of diseases of the internal organs. A quantitative scale assessing the state of damage to the tissue of internal organs allows you to more accurately determine the levels of pathological disorders occurring inside this tissue. The method of obtaining substances is technically simpler, has a lower cost per unit of time during production, is safer in production, since it eliminates radiation sources of radiation, is more accessible, allows conservation and long-term storage of the source material, includes the steps of disinfection of the intermediate product, which makes it safe for personnel to work with.

The claimed technical solutions are based on the creation of a new class of substances with immune activity, derived from the tissue of an internal, transient or transplanted organ, a neoplasm (tumor) of an individual's organism, possessing specific activity against antibodies / autoantibodies and characterized by the presence of properties that allow to detect the state of the tissue using a laboratory method the internal organs of the individual being examined by detecting the antigen-antibody reaction, an autoantigen-autoantibody, that occurs between tage and biological fluid withdrawn from the subject's individual having partial syngeneic organ and species, which allows to cause the immune response system of the body of the individual by parenteral administration, resulting in local tissue lysis having organ and species tropism for injectate.

The inventive substance has new properties, manifests itself as a glycoprotein, provoking the formation of antibodies inside the body when administered parenterally, which are immunoglobulins (M, C), and capable of forming antigen-antibody complexes (autoantigenautoantibody) with anti-C anti-M immunoglobulins. Moreover, antibodies produced against the substance, due to the property of partial organ syngene, are anti-tissue and anti-organ autoantibodies of the living tissues of the body. Such properties of the substances described in the analogues, no. Tissue of any object of the animal world cannot be used as a raw material for obtaining a substance, and any tissue found in a living organism, in an inanimate organism, in a remote organ, in punctate, in a transient organ, if there is no tissue signs of biological lysis (decomposition) and there is the possibility of typing and excretion of at least 1000 mg of the original tissue.

In the claimed invention, a group of substances has been obtained that have new properties based on the organ and species partial syngeness of the original tissue, which allows the resulting substance to be used as the basis for diagnostic and pharmaceutical agents used simultaneously. When a substance is used as a pharmaceutical agent, its action is realized as a vaccine, which is capable of causing a cascade of physiological reactions in which the body’s immune and autoimmune systems are involved, leading to local isolated lysis of the tissue inside a living organism that does not lead to its death. When using the same substance as a diagnostic tool at the time of treatment (vaccination) of an individual with the same substance, it is able to detect the magnitude of the body’s response to its administration (detect the magnitude of the substance being vaccinated), engaging in immune responses in the antibody autoantibodies formed in the body response to its parenteral administration. These properties of a substance allow a personalized, individualized treatment — when the individual can be the raw material for the production of a substance. The proposed method of obtaining the substance and its application allows the use of trace amounts of the source material, which is extremely important for the implementation of the principles of minimally invasive treatment.

A quick and accurate diagnostic result (up to 90 minutes), immediately predicted level of health of the tissue of internal organs and the estimated state of health of the body during the next 24-48 hours, detection of latent health disorders or damage to internal organs, easily accessible and simply realizable possibility of a diagnostic technique allow apply this method to identify possible complications and adverse reactions when conducting mass vaccinations with other vaccines and to carry out the selection of patients for these purposes.

The technical result also consists in the specific effect on the isolated original tissue of a biological individual, which allows preserving the organ and species specificity of the original tissue, leveling its allogeneic characteristics, and obtaining a new substance from the partial organ and species syngenostyo to the original tissue; obtaining a new substance with other properties allows it to be used for early direct diagnostics of the state of the tissue from which it was obtained, using it as part of a laboratory test, and locally lytically affecting the tissue, using the substance as part of a pharmaceutical composition for parenteral administration, using the organ property and species singularity of a substance devoid of allogeneic properties for precise, local controlled lysis of the same tissue, while simultaneously monitoring the result and step This tissue is affected by the same substance included in the laboratory test used to control treatment. Controllability and accuracy of local exposure to a substance, minimizing the harmful and toxic effects that inevitably occur when a local lysis of the tissue inside a living organism, allows you to personalize and individualize the treatment in each case, to select an individual dose of the substance (the amount of the substance; the amount, time and need for repeated administration of the substance ), not leading to death; in the implementation of laboratory control of the treatment carried out by this substance, its ability to detect antibodies-autoantibodies in biological samples during treatment due to the fact that the substance has a high sensitivity, specificity, which are due to the properties of its partial organ and species syngene to an identical tissue, from which it is received and able to detect

- 5 018848 minimal changes (lytic damage) of the tissue, which do not lead to incompatible with life consequences inside the organism, significantly without changing its general condition, accurately dosing and evaluating the necessary levels of lytic influence on the tissue.

The task is solved by the fact that the immunologically active substance (IAV) is a protein substrate obtained from the supernatant of a modified original organ tissue of an individual, characterized by partial syngene, having full organ and species antigenic similarity to the original tissue, lacking the allogeneic antigenic properties of this tissue, exhibiting properties and antigen and autoantigen to the tissue from which it was obtained, and having specific activity against antibodies-autoantibodies that are in biological FIR liquids other individuals of the same species. The immunologically active substance contains macromolecules of the protein substrate in the molecular weight range of 10–90 kDa, in which, during denaturing electrophoresis, specific areas are detected in the form of at least two major bands corresponding to the values of the molecular weight scale between 9.0 and 11.0 and 59 61 kDa.

The immunologically active substance contains at least one immunoactive syngeneic epitope with antigenic-autoantigenic properties of the original organ and species of an individual.

The combination of the properties of an iAv is determined by the formula

C = ⁽ Σν (+ Σνν + ^ι Σθνί + ^η Σ _8νν where ^ι Σ _ν1 - the organ aggregate of similarity, the aggregate of substances that determine the similarity of tissue according to organ features (νΐ), ΐ is the type of tissue;

Σ _νν - species set of similarity, a set of substances that determine the similarity of the tissue according to species characteristics (νν), η is a type of an individual;

ΐχ- 'o

Σανΐ - organ combination of differences, the totality of substances that determine the differences of tissue on the basis of organ belonging (ανΐ), ΐ - type of tissue of the organ;

η

Σ _{; ινν} - specific set of differences, the set of differences of substances that determine the differences of tissue on the basis of species (ανν), η - the type of individual.

Immunologically active substance is characterized by the presence of two or three dominant fractions of alkaline components when performing high-performance liquid chromatography, weakly interacting with the anionic column at pH 7.2, and differing in charge acidic major components eluted in the middle and at the end of the chromatography process, and which appear on chromatography curve in the form of peaks corresponding to the ranges of 33.5 ± 2.5 and 52.5 ± 2.5 mAy for the two dominant fractions of alkaline components or the corresponding range am values 17 ± 2,5; 33.5 ± 2.5; 52.5 ± 2.5 for the three dominant fractions of alkaline components.

The method of obtaining an immunologically active substance includes the collection of the original tissue, its typing, repeated freezing at a temperature of (-24) - (- 28) ° C and thawing, while thawing is performed by exposure to microwave radiation at a frequency of 2450-2500 MHz with removal of defrosting moisture to reduce the weight of the original tissue by 30-60%, the resulting tissue resulting from such processing, characterized by a syngality to the species and organ antigenic codes of the original tissue and lacking the allogeneic antigenic properties of the original tissue and Ndivida, crushed, homogenized, disinfected and extracted according to Grasse, the resulting supernatant is subjected to standardization of protein from 20-25 mg per 1 ml and freeze-dried. Tissue disinfection is carried out by adding 0.5% aqueous formalin solution to a tissue homogenate to 5-10% of the total starting material, followed by mixing and exposing the mixture for 4-6 hours. The microwave effect is carried out at a radiation source power of 600-1000 W for time, which is set at the rate of 10-50 s per 100 g of the mass of the original tissue, while in the process of microwave exposure, these parameters are set based on the condition of preventing heating of the original tissue to a temperature of 41 ° C. Microwave exposure is carried out during the time at which the water crystallized on the surface of the frozen tissue becomes liquid.

An immunologically active substance is used as a marker of the state of the tissue of the internal, transient, or transplanted organs of an individual.

A bioprobe can also be manufactured, including a solid phase with an immunologically active substance applied onto its surface. The immunologically active substance is applied to the solid phase of the bio-probe as a series of dilutions in the concentration range from 12.5 to 200 μg of dry matter per 1 ml of physiological solution. A series of immunologically active substances obtained from various tissues of an individual's organs can be applied to the solid phase of a bio-probe.

A method for diagnosing the condition of a tissue of an internal organ of an individual includes taking a sample of a sample of a biological fluid of an individual, ensuring contact of the biological fluid with an IAA according to claim 1 (see claims) or a biosonde according to claim 11 in a series of dilutions of the sample of the test fluid within the titer values from 1 : 4 to 1: 2048, determination in titers of the interactions between antibodies and / or autoantibodies found in a diluted sample of an individual's biological fluid and an immunologically active substance according to claim 1, with a conclusion about The tissue of the internal organ of the individual is made according to the marginal positive result of interaction in a series of dilutions

- 6 018848 sample of the test liquid, while, if the interaction is detected within the dilution of the test sample within the values [1: 4-1: 8], it is concluded that the tissue is healthy; within the range of [1: 16-1: 32] - concludes that there is insignificant tissue damage, within the range of [1: 64-1: 128] - they conclude that there is damage to the tissue typical for chronic inflammation, within the range of [ 1: 256-1: 512] - concludes that there is damage to the tissue that is typical for acute inflammation, within the range of [1: 1024-1: 2048] - they conclude that there is damage to the tissue that is typical for lysis and / or rejection of the tissue.

The pharmaceutical composition is a solution comprising an immunologically active substance according to claim 1 and an acceptable solvent, while for parenteral administration 1 ml of the solution contains 100 μg of the substance.

A method for the treatment of diseases of the tissues of an individual's organs caused by uncontrolled growth of malignant or benign nature tissue cells, includes administering to an individual parenterally by uncommon the pharmacological composition according to claim 15, containing the substance according to claim 1, starting with a dose of 50 μg of the substance and then increasing to sensitization of the body and obtaining a response of the immune system of the body of the individual, which is judged by the formation of anti-tissue antibodies, autoantibodies to the injected substance according to claim 1, however, the introduction of the pharmacological composition is carried out by monitoring the level of formation of antibodies-autoantibodies in the biological fluids of the patient, which are taken after each administration of the composition; the number of injections is limited when reaching the level of antibody titers-autoantibodies lying in the range of values from 1: 256 to 1: 512.

Brief Description of the Drawings

The group of inventions is illustrated by drawings, which show electron diffraction patterns of the ultrastructure of the liver tissue of animals in the control sample and in the experimental group, namely:

in fig. 1 shows the electron diffraction pattern of guinea pig liver ultrastructure in a control sample with magnification x8000;

in fig. 2 - electron diffraction pattern of the local site of damage to the liver of a guinea pig (the result of the substance's exposure to the local portion of the liver tissue of the guinea pig), which arose after the termination of the claimed substance, an increase of 7500

in fig. 3-6 - electron diffraction patterns of ultrastructure of liver tissues of guinea pigs exposed to the test substance, one of which fell after 100 hours of the experiment, an increase of x8000 (Fig. 3), the other — 72 hours after the start of the experiment after repeated administration of the test substance, an increase of x5600 ( Fig. 4), the third - 72 hours after the start of the experiment after re-injection of the test substance, an increase of x8200 (Fig. 5), the fourth - 132 hours after the start of the experiment, and received 2-fold administration of the test substance, an increase of x8000 (Fig. 6);

in fig. 7 presents the results of HPLC (high performance liquid chromatography) of the substance obtained according to method 1 from human liver;

in fig. 8 - HPLC results of a substance obtained according to method 1 from human kidneys;

in fig. 9 - presents the results of electrophoresis of samples of the claimed substances according to claim 1, obtained by the method according to claim 6;

in fig. 10 - results of immunodot analysis;

in fig. 11 - the results of ELISA (enzyme-linked immunosorbent assay), showing the dependence of optical density values on the dilution of the antigen (substance - the liver), at a wavelength of 490 nm;

in fig. 12 - the result of the reaction of rnga, demonstrating the possibility of detecting anti-tissue autoantibodies by a number of substances obtained from tissues of different organs, in dilutions of human serum.

Causal relationship of the signs of the formula with the achieved result

For disclosing the mechanism of the influence of new features identified in the substances obtained by this application, allowing them to be used according to the claimed formula, confirmed by the course of further research and experiments. Below is the course of the system analysis with the formal calculations that allow to realize them practically and transfer the results to the claimed object. The theoretical background and system analysis of the antigenic sites of tissue proteins, which have the ability to show partial syngene, are also given.

Antigens that are unique to this organism are called alloantigens (Greek allos is another). These include antigens of tissue compatibility - the gene products of the main complex of tissue compatibility MHC (Ma_) og SCHSootryPyu Comparty), peculiar to each individual. Antigens of different persons that do not have differences are called syngeneic. Organs and tissues, in addition to other antigens, possess organ and tissue antigens that are specific to them. The antigenic similarity has the same tissue of biological species, for example, humans and animals. There are stage-specific antigens that appear and disappear at certain stages of the development of tissues or cells. Each cell contains antigens characteristic of the outer membrane, cytoplasm, nucleus and other components. Incomplete antigens are found in nature quite often. Incomplete antibodies detected

- 7 018848 us against Escherichia, Brucella, Salmonella and other microorganisms; against red blood cells and other cells. According to the classification of antigens distinguish exogenous (coming from the outside of the body) and endogenous (formed inside the body) antigens. For the case under consideration, endogenous antigens were selected.

Antigens are divided into complete and incomplete (haptens). The complete antigen includes a carrier protein and a determinant group, they can induce the formation of antibodies and lobby with them. The property of an incomplete antigen lacking a carrier protein is known to react with antibodies, but these substances cannot stimulate their formation inside the body. Incomplete antigens (including haptens), entering the body, can combine with the body's protein and become complete antigens.

The antigens of each organism do not normally cause immunological reactions in it, since the organism is tolerant to them. However, under certain conditions, they acquire signs of foreignness and become autoantigens, and the reaction that occurs against them is called autoimmune.

Malignant tumor cells are variants of normal body cells with an antigenic structure that is virtually no different from the body’s antigenic structure. They are characterized by antigens of those tissues from which they originated, as well as antigens specific to the tumor and constituting a small fraction of all antigens of the cell. During carcinogenesis, dedifferentiation of cells occurs, therefore, some antigens may be lost, and antigens characteristic of immature cells may appear, including embryonic (fetoproteins). Antigens that are unique to a tumor are specific only to this type of tumor, and often to a tumor in a given person. Tumors induced by viruses may have viral antigens that are the same in all tumors induced by this virus. Under the influence of antibodies in a growing tumor, its antigenic composition may change.

The protein structure of an individual contains at least three relatively stable and informationally significant antigenic regions that encode species, individual and organ specificity. Recognition of protein foreignness inside the body occurs by assessing areas of protein structures that carry information about the organ, species and individual compatibility of the tissue.

The analysis of the data obtained according to the claimed invention and the analysis of body reactions confirmed the presence of at least three sites encoding the species, organ and individual features in the structure of the protein. The author proved that changes in these areas allow obtaining new substances that are able to detect organ and species-specific areas of protein structures in biological fluids of individuals, entering into immune reactions with them, and detecting and marking identical tissue by the inner link of immunity, leading to its lysis in the case of receipt of this substance in the body. The result of the research proves that the complex structure of the protein substrate carries information about who owns this protein object (human, animal, or belongs to the world of simple organisms). In addition, the protein has information about which tissue it belonged to, since it contains information about its organ affiliation. The protein also carries a unique individual code that allows you to accurately recognize exactly which individual it belongs to.

The theoretical foundations and the course of the analysis were accompanied by the introduction of the following system statements giving grounds for its implementation.

The organ aggregate of similarity (Σ ^) is the similarity of functionally identical organ tissues, where (νί) are signs of the organ similarities of these tissues. For example, liver tissue is functionally similar in animals and humans, as it performs similar functions. The liver tissue is morphologically similar, since it can be structurally divided into hepatocytes - 60-70%, bile ducts and paths - 1218%, vessels - 20-25%, stroma - 2-3% in humans and animals, and the percentage the structures will vary with minor errors from the main representation of the structure of the cells entering the tissue of the organ. All tissues of the same type organs in different species are united by a single anatomomorphological structure, physiological and functional purpose. The same reasoning will be valid for the tissues of other organs: the stomach, intestines, brain, heart, glands, etc.

Species aggregate similarity (Σ _νν ) - the similarity of the structure of cells, tissues and organs in individuals of the same species, and (νν) - signs of the species similarity of these tissues. Interspecific similarity of the structure and functions of the liver, heart, lungs, glands, etc. between people, between dogs, between horses, etc.

Individually-typological set of similarity of cells within a single organism (Σ _νί ) - the similarity of protein structures of various organs within one single living organism, where (νί) - signs of individuality of tissues in one organism.

Based on the introduced similarity sets, the informational image of the tissue will be a component of the entered sets and can be represented by the formula in the form:

where C is the informational image of the tissue, based on the aggregates of their similarity.

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Difference sets (differences, differences):

Species collection of differences (Σ _ανν ) - differences in the structure of tissue in individuals of various species, with (ανν) - signs of species differences in the compared tissues, for example, differences in the structure of man and horse.

The body of difference (Σ _ανί ) - differences in morphological, structurally cellular, tissue and functional structure of different organs, and (ay!) - signs that define organ differences, for example, liver tissue differs from the tissue of the stomach, heart, etc.

Individual set of differences (Σ _ανι ) - differences in the individual characteristics of organisms of individuals of the same species, where (ανι) are signs of individual differences between identical tissues in individuals of the same species, for example, differences in individuals by blood group, pigment amount, etc.

Based on the introduced sets of differences, the informational image of the tissue will be a component of the entered sets, and it can be represented by the formula in the form:

where C is an informational image of the fabric, based on the totality of their differences.

Of course, you can enter another set of similarities and differences between the analyzed objects on other grounds (species, sex, age, etc.). However, these aggregates do not have a significant impact on the subject of analysis, cannot significantly affect the general course of reasoning, and therefore their values were excluded from the analysis.

The following is a systematic analysis of informational images and the typing of randomly selected tissues of three objects: (the first object is a guinea pig (liver), the second object is human I. (liver), the third object is human P. (liver). By leveling in the analysis the significant differences in the characteristics of these tissues (structural, morphological, functional, etc.), the species and individual populations of interest are selected and combined according to signs of similarity and differences. In the course of mathematical modeling of the informational image of the tissues of objects, during the analysis, comparisons of similarities and differences of objects are used, which are described by the above formula. Below are the resulting conclusions and the following of them postulates (without intermediate reasoning).

The first postulate. The liver tissue of an animal (guinea pig) is similar to human liver tissue (the first object is similar to the second and third objects), since the liver of all the objects studied has similar functions and has a fundamentally similar structure in all mammals. A common pattern is the similarity of the structure of protein structures of tissues of similar organs in different species and within each species - the similarity of organs in terms of the totality of features, functional load and structure in individuals between and within the species.

Thus, the three analyzed tissue objects have a common similar set, similar in terms of features reflecting their functional purpose and the structure of organs. Fabrics have a certain common feature - a common information code, reflected in the protein structure and characterizing its essence - the liver.

The second postulate. The differences in the tissues of the three objects studied are available by species code. Moreover, tissue samples (liver) of the first object differ from the samples of the second and third objects (differences in the structure of guinea pig and human organisms, individuals I. and P.). The biological objects analyzed by us (the second and the third) belong to the same species - the man, and the first object belongs to the other species - the animal. General pattern - differences in protein structures of tissues of organs in different species.

Thus, the analyzed objects have a common set of differences of organs and tissues, species differences between the objects. Fabrics of objects are separated by a species information code inherent in each species of living fauna, in our example, man and animal. Protein structures of tissues within species are endowed with such differences, the separation bears informationally significant differences of a typed object according to the species of the tissue. Protein structures of tissues of a single species, when ingested into another species, will be identified and destroyed by the autoimmune system as alien by species.

The third postulate. When comparing the three samples of the analyzed tissues of the liver, animals and people, they were found to differ from the tissues of other organs (tissue of the stomach, heart, lungs, glands) in functional purpose, structure, etc. This is an axiom that does not require proof, which means there is an aggregate differences in protein structures of cells of organs and tissues within a single organism.

Thus, the analyzed objects have a common set of differences of organs and tissues, the differences of tissues between the organs of one organism. Organ tissues are separated by organ information code, inherent in each organ and tissue within the body. Protein structures of tissues and organs within each living organism are endowed with such differences, the separation carries informationally significant differences of a typed object by tissue and organ affiliation. Protein structures of tissues of various organs, outside their cell pool, can be recognized and destroyed by the autoimmunity system.

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The fourth postulate. Two samples — liver tissue I. and liver tissue P. — have similar species and organ characteristics, but have intraspecific and organ differences according to an individual-typological trait. Macro-organisms and individuals I. and P. differ in the totality and set of genetic and informational characteristics inherent in each of the individuals separately. There are similarities in the characteristics of the species and organ codes of the samples and the totality of differences between the protein structures of the organs according to unique individual-typological characteristics, each individual, within a single biological species or subspecies.

Thus, the analyzed objects have a common set of similarities between the intraspecific and organ-tissue protein structures, but there are differences in tissues at the level of individual typological differences between organisms. Organ tissues are separated by a unique individual-type code, which is inherent in each organism separately. All protein structures of tissues and organs within each individual, each individual within each biological species or subspecies, regardless of its position on the biological hierarchy, are endowed with such differences. The division carries informationally significant differences in the protein structures of the type of objects according to the unique individual genetic code, which allows to determine whether or not its protein structure is inside the body. The body identifies such differences and can reject such objects, despite the similarity of the species and organ codes of protein structures (graft rejection reactions).

Fifth postulate. If you compare the three samples of the analyzed tissues (liver) with a sample of tissue of the stomach, as was done in the third postulate taken from one of the studied organisms, it is possible to isolate one more important detail. Based on the supplement we introduced that a sample of stomach tissue could belong to or belong to one of the analyzed organisms, despite the difference in structural differences and protein codes characterizing liver tissue and stomach tissue, with organ or species difference between samples, one of the individuals that to whom it belonged earlier and was subsequently collected for research, the tissue will not be alien. It has its own despite the difference between the organ protein code. It has its own individual-type protein code and the species code to which this organism belongs. This is a common informational individual-typological code of the protein structure of tissues of various organs inside one organism. Such tissues, having a different organ code, are not differentiated by the autoimmunity system as strangers, are not destroyed and not utilized, despite the existing organ difference and the dissimilarity of the functional purpose of the original protein substrate.

Thus, the analyzed objects have a set of differences of organs and tissues, differences in the tissues of the organs inside one organism, in all three analyzed cases, but in one case the tissues, having differences in organ code, have a single unique individual-typological code inherent in one organism. Organ tissues are separated by an organ information code, inherent in each organ and tissue; if they are inside one organism, then they have a matching unique individual-typological species code belonging to this organism. Protein structures of tissues and organs within a single living organism are endowed with such similarities, the separation carries informationally significant differences of a typed object by tissue and organ belonging, but it has a completely identical unique organism code — one host and is not recognized by the autoimmunity system as someone else’s, does not affect their rejection and not destroyed.

Thus, the organism reacts to a suddenly reborn, mutated under the influence of an external protein structure, leading to the formation of tumor cancerous tissue. It is this fabric, not alien, it is its own, its unique individually typological code, the code of the host-individual is preserved and coincides with it. The changes have affected the organ - structural code of the tissue, the tissue has become a foreign organ, the functional purpose of this tissue has changed, which no longer allows the cell to perform its former functions, and such (not rejected or not utilized) cell begins to be intensively maintained and developed inside it organism. A new process begins, the growth and development of such its own tissue increases, which, not knowing the boundaries of tissue and organ pools, over time, takes over the entire host organism, causing its inevitable death.

Our analysis allows us to draw the following conclusions: an antigenic image that defines an informational mosaic of tissue features (C) can be represented by a mathematical expression using the analysis of the totality of signs of similarities and differences in tissue.

Using the proposed formula, we can describe the liver tissue (ΐ) of a specific (Ν) guinea pig (p). If the tissue belongs to one type (p), one type (ΐ) of tissue and one organism (Ν), then the formula of the information image of the tissue will have the following form:

= ^one U 4- ^P 4- ^ 4- * 4- ^P U 4- ^ U <Α ^ νί '4-νν' 4-νί '- “ _aU 1 'A, _ayy T A. _ay | Where C is an informational image of a tissue, based on the totality of their similarity and the differences in their characteristics inherent in one type, one type of tissue found in one organism.

If we have another fabric with a formula that can be described in a similar way, then the shape

- 10 018848 mules of the information image of the tissue will have the following appearance:

c = * y + y + ^ y -4- ^one y + ^P y + Ά where C is an informational image of another tissue, based on the totality of their similarity and the differences in their characteristics inherent in one type, one type of tissue found in the body of a particular individual.

Thus, the considered mathematical expressions characterizing tissues are identical and completely identical, therefore, the informational images of the compared tissues, characterized by these expressions, are identical, and therefore, the antigenic sets of signs of the tissue - the syngeneic tissues - also coincide.

If we consider another formula of an information image of the fabric, having the form

P = ^eleven Y-4- ^P U 4-. four- ^G1 U 4- ^P In 4ν ν ('νν' νι 'a \'('"νν' νΐ, where 11 is the tissue of another organ, we can state, using the similar reasoning model given above, that the characteristic of aggregates of organ differences and similarities is changed (νΐ , a \ '1), which means allogene tissue to a given organism.

If we consider another formula of the information image of the fabric, having the form

C = ^ι1 Σν1 + Σνν + ^ι 'Σ3ν1 +% _νν , „ ^Ν Σ3νϊ and ^Ν Σνί,.

where the values of the sets are changed (the signs of individual similarities and differences are neutralized) so that the values in the formula can be equated to a zero value, then we can assert that this is an informational image characterizing another substance and this mathematical expression refers to another substance having complete sets sets of organ and species signs of similarities and differences of tissue, but devoid of signs of individuality of a particular organism. It can be argued that the substances that determine the antigenic characteristics of such a substance have a complete coincidence of sets of similarities and differences in the characteristics of the organ and species characteristics of the original substance obtained from the tissue of a particular individual, i.e. there is a partial (partial) syngeness of the described tissues on the basis of a set of signs of organ and species similarity and differences. A substance having such an initial formula is not identical to the one being compared, it is another substance.

Thus, it is possible, based on descriptions of sets of similarities and differences in tissues, to bring the conclusions of system analysis to general practice and conclude that:

1) there are antigenic differences in the structure of protein structures of tissues of individuals of various species;

2) there are antigenic differences between the protein structures of the tissue of organs inside the body;

3) there are antigenic differences in the structure of the protein structures of the tissues of individuals of the same species. They can completely neutralize the similarity of the structure of the protein structure of this species as a whole;

4) the unique individual-typological antigenic code of the organism present in each protein structure of the tissue of the whole organism eliminates the difference in protein structural differences according to the organ feature, making all the protein structure codes that do not coincide on the basis of tissue unity and function for the autoimmunity system.

The correctness of the conducted system analysis and the above logical tenets is confirmed practically.

The inventive method allows to obtain substances that can be described by the formula

C = * Σ _νί + ^η Σνν + ^Ν Σνί + ^ι Σ8νί ⁺ Σ3νν + ^Ν Σ _Ην ί, where C is an antigenic set of signs of a substance that determines the informational image of the tissue, based on the sets of signs of similarity and differences;

^ι Σ _ν1 - the organ aggregate of similarity, the aggregate of substances that determine the similarity of the tissue according to the organ features (νΐ), where ΐ is the type of tissue;

^η Σ _νν - specific set of similarity, set of substances that determine the signs of similarity of tissues within a single species (νν), where η is a type of an individual;

Νρ

Σ _ν ί is an individual totality of similarity, a totality of substances determining the signs of tissue similarity according to individual-typological characteristics (νί), where N is an individual;

^ι Σ _3ν1 - the organ combination of differences, the totality of substances that determine the signs of differences in tissue according to organ affiliation (a \ '1);

^η Σ _3νν - species set of differences, the set of differences of substances, which is determined by signs of differences in tissue by species (ayy);

Νν

Σ _3ν ί - an individual set of differences, a set of substances that determine the signs of tissue otlinia according to individual typological characteristics (ay1).

The inventive substances are determined by the antigenic set of characteristics of the original tissue, but meet the requirements of the formula

C = 'Σνΐ + Σ _νν + 'Σ ₃ νί + ^η Σ _3νν in which the values of individual sets of signs of similarity and differences of the original tissue

- 11 018848 neither ^χ Σ; ινι and ^χ Σνι modified by the claimed method, do not show their properties and have values equal to zero, while maintaining the full values of the sets of signs of similarities and differences of the original tissue by organ and species characteristics. In the claimed substances, the individual-typological properties of the initial substance are leveled (minimized) as far as the properties of these substances can determine the signs of individual-typological similarities and differences of this tissue in a living organism, which is able to recognize them as its own.

The essence of the group of inventions is to obtain new immunologically active substances (IAV), exhibiting new properties that can be described by the formula

C = ⁽ Σνϋ + ^η Σνν + 'Σ _{3ν (} + ^η Σ _3νν , and which form the basis for the diagnosis of the state of tissues and their treatment. The whole group of substances that can be described by the formula of the totality of similarities and differences can be used for the purposes of this technical solution.

All IAV, obtained by the claimed technology, retain a combination of signs of organ and species similarity and tissue differences (organ and species syngeness), the tissue from which they are derived, and can simultaneously exhibit the properties of a direct tissue antigen or direct autoantigen to an identical tissue depending on methods of its use and application. In laboratory reactions, IAV exhibits the properties of direct autoantigen of identical tissue, a marker of which it is, and is capable of detecting the level of anti-tissue autoantibodies in the biological fluids of an individual. In the organism of the same species, when administered parenterally, IAA exhibits the property of a direct antigen of identical tissue, a marker of which it is, leading to a cascade of immune and autoimmune reactions that lead to the development in response to its introduction into the body of antibodies, which are at the same time autoantibodies to identical tissue located in the living organism of the individual, leading to mediated damage and lysis of this tissue in the living organism of the individual due to the impact on it of the formed antibodies-autoantibodies.

The described immunologically active substance (IAA) has the properties of an incomplete tissue antigen of the individual from which it is derived.

The claimed variants of the substance belong to the biotechnologically created substances, they are protein-like, capable of inducing the formation of highly specific anti-organ anti-tissue antibodies (autoantibodies) when administered parenterally, not being the complete antigen of the original tissue from which it was derived. In his code there is no area responsible for the individual compatibility of the fabric. At the same time, due to the syngeneicity of the organ and species code of antibodies — autoantibodies of an individual donor or an individual of the same species, substances are capable of entering immune reactions with it under laboratory conditions. Thus, the substance has a complete set of codes responsible for the organ and species affiliation of the antigen (it is syngeneic to the original tissue according to the set of organ and species traits), but it is different from the fact that the substance codes lack a set of signs of allogeny - individual histocompatibility compatibility. The inventive method of obtaining the UAB ensures the achievement of the above characteristics.

Obtained by us IAV exhibit antigen and autoantigen properties in various conditions of their use. IABs acquire allogeneic properties and are identified by the immune system of an individual of the same species, or by the immune system of a tissue donor organism, as foreign antibodies and antibodies produced by antibodies that can destroy identical tissue are produced against them.

IAA in laboratory reactions exhibit autoantigen properties and are able to detect the level of anti-tissue autoantibodies in biological fluids of an individual, and they are also able to exhibit the properties of a direct tissue antigen when parenterally administered to the organism of the same species, leading the body’s response to a cascade of immune and autoimmune transformations that lead to development in response to its introduction into the body of antibodies, which are at the same time autoantibodies to identical tissue found in the living organism of the individual. It is the increasing level of autoantibodies of the individual that leads to damage and lysis of the antigen-identical (organ-syngeneic) tissue in the living organism of the individual.

The implementation of the invention

Substances by the present method can be obtained from the tissue of any organ (except blood) of an organism of any kind, from the tissue of the transient organ (placenta), from the modified (tumor), but intact developing tissue from the individual (donor) and used as a diagnostic substance or composition of the pharmacological composition, in the same individual or individuals of the same species, for diagnosing the state of health of the tissue from which it was obtained, for treating (by lysis) a portion or all of the tissue in the body for monitoring the degree of tissue lysis and evaluating the results of treatment with the same substance.

In more detail the steps of the method of obtaining the AAA disclosed in the table. one.

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Table 1

Nn Technology stage, object Raw material processing method and modes one Intake and pathological examination of material, tissue typing Sectional material, organ tissue obtained by biopsy, tissue of the organ removed during surgery, tissue of the transient organ (placenta), graft tissue, organ tissues remaining after histological and pathological studies. 2 Preservation - freezing and storage Freezing at a temperature of -24-28 ° C for at least 24 hours, storage for no more than 30 days. 3 Defrosting and thawing using microwave Microwave - exposure at a frequency of wavelengths in the range of 2450-2500 MHz, the power of the radiation source in the range of 700-900 W, with a duration of 10-30 seconds per 100 grams of material four Dehydration, removal of moisture and liquid Draining, drying (wetting woven). 5-6 Additional double repetition of cycles 2, 3 and 4 Repeating 2, 3 and 4 cycles 7 Homogenization of thawed tissue Grinding, homogenization and extraction according to Grasse eight Disinfection of the homogenate extract Add 0.5% aqueous solution of formalin to 5% of the total extract, exposure for 4 hours 9 Fractionation of the extract Centrifugation of 22,300 c for 15 min. ten Sediment removal Removal of moisture eleven The selection of the supernatant Standardization of protein 22 mg / ml 12 Supernatant processing Lyophilic drying 13 Obtaining the target substance Storage

Examples of obtaining substances according to claim 1 of the formula from the tissues of various organs and species are given in table. 2

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table 2

Cycle 1 Cycle 2 Cycle 3 Cycle 4 Cycle 5 and & Cycle 7 Cycle 8 Cycle 9 Cycle 10 Fabric intake Freeze storage with Microwave effect Dehydration Freezing, microwave exposure, dehydration Homosexual Disinfecting live Getting supernat nta Obtained target substances Type and weight of fabric temperature T ° and time of freezing мо and bookmarks and on preservation Wavelength Power, Duration of Radiation Full defrosting at a temperature of 25 ° С and removal (draining) of the image of the injected liquid and Additional double repetition of cycles 2, 3 and 4 Rubbing and grinding with quartz sand (gr.) With 0.9% NaCl solution (ml) until complete homogenization Introduction of 0.5% pp formalin and 4 hours of exposure to a homogen Centrifugation 22300 ς for 15 min., Selection of supernatant, protein standardization to 22 mg / ml Amplification, labeling, lyophilization, packaging and storage Smooth muscle tissue of the stomach of a corpse of a rabbit, 10 grams. -28 ° C, 54 hours 2450 MHz, 800 W, 10 sec. Removal of 1.5 ml. thawed liquid and after 30 min. ottai wa niya -28 ° C, 24 hours, 2450 MHz, 800 W, 10 sec. Twice but removed 2.3 ml. thawed liquid and 5.8 grams of tissue + 10 grams of sand + 5 ml. 0.9% №С1 within 2 hours Introduction of 0.75 ml of 0.5% pra formally in the homogen T for 4 hours Centrifugation for 15 minutes, selection of 9, 2 ml of the supernatant, determination of protein in the liquid and addition of 3 ml. 0.9% Cac1 to 22 mg / ml protein in supernat nte Spill into labeled ampoules of 3 ml, lyophilic vacuum and dry drying, sterile sealing of substances and storage in ampoule (2 ° C)

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Smooth muscle tissue of the stomach of a human corpse, 35 grams. -24 ° C, 96 hours 2450 MHz, 900 W, 22 sec. Removal of 4.5 ml. thawed liquid and after 30 min. defrosting -28 ° C, 24 hours, 2450 MHz, 900 W, 22 sec. Twice but removed 6.7 ml. thawed liquid and 21.9 grams of cloth + 50 grams of sand + 10 ml. 0.9% YaC1 for 3 hours Introduction of 2.5 ml of 0.5% pra formali in the homogen T for 4 hours Centrifugation for 15 minutes, selection of 18.0 ml of the supernatant, determination of protein in the liquid and addition of 6.4 ml. 0.9% 1CHaC1 to 22 mg / ml protein in supernat nte Spill into labeled ampoules of 3 ml, lyophilic vacuum and dry drying, sterile sealing of substances and storage ampoule (2 ° C) Human liver cadaver tissue, 62 grams -24 ° C, 38 hours 2500 MHz, 700 W, 30 sec. Removal of 9.5 ml. suckers after 30 min. defrosting -24 ° С, 24 hours, 2500 MHz, 700 W, 30 sec. Twice but removed 18.7 ml. sods 39.9 grams of cloth + 70 grams of sand + 15 ml. 0.9% №С1 within 3 hours Introduction of 3.1 ml of 0.5% pPa formally in the homogen T for 4 hours Centrifugation for 15 minutes, selection 34, 0 ml of the supernatant, determined protein in liquid and added with 9.7 ml. 0.9% of number C1 to 22 mg / ml of protein in supernat nte Spill into labeled ampoules of 3 ml, lyophilic vacuum drying, sterile sealing of substances and storage ampoule (2 ° C) Human cadaver tissue, 42 grams -24 ° C, 86 hours 2500 MHz, 700 W, 20 sec. Removal of 6.5 ml. suckers after 30 min. defrosting -24 ° С, 24 hours, 2500 MHz, 700 W, 20 sec. Twice but removed 10.2 ml. sods 27.1 grams of cloth + 20 grams of sand + 10 ml. 0.9% Iac1 for 2 hours Introduction of 2.0 ml of 0.5% pra formali in the homogen T for 4 hours Centrifugation 15 minutes, selection 23, 5 ml of the supernatant, determination of protein in the liquid and addition of 4.7 ml. 0.9% 1MaC1 to 22 mg / ml Spill into labeled ampoules of 3 ml, lyophilic vacuum drying, sterile sealant

- 15 018848

Corpse -24 ° C 2450 Remove -24 ° C 23.2 Introduction protein in supernat nte centrifu substances in ampoule storage e (2 ° C) the cloth 120 Mhz, e 4,5 24 hours, gram 2.15 ml giro marco white o'clock 700 W, ml. 2450 fabrics + 0.5% p- 15 minutes, the bathrooms substances 10 sec thawed Mhz, 50 ra selection 17, 0 ampoules headache fluid 700 W, gram formals ML on 3 ml, about the brain and after 10 sec sand + on in supernatant lyophile human 30 min. Double 18 ml. homogenous is defined a little 31 grams ottayva but, 0.9% t on 4 squirrel in vacuum n of deleted 1CHA1 in hours liquids oh 7.7 ml. flow AND dry out thawed 3 hours adding the knowledge fluid e 11.1 ml. sterile and 0.9% Iac1 Naya up to 22 sealing mg / ml ation squirrel in substances supernat and in nte ampoule storage (2 ° C) Operations -24 ° C 2450 Remove -24 ° C 3.2 Introduction Centrifuge Spill into onny 96 hours Mhz, e 1,2 24 hours, gram 0.3 ml giro marco material 700 W, ml. 2450 fabrics + 0.5% p- 15 minutes, the bathrooms fabrics 10 sec thawed Mhz, ten Ra screening 3.1 ampoules parenhi fluid 700 W, gram formals ml on 3 ml, we are testicles and after 10 sec sand + on in supernatant lyophile human 30 min. Double 3 ml. homogenous is defined a little 7 grams thaw but, 0.9% t on 4 squirrel in vacuum of deleted №С1 to hours liquids oh 1.7 ml flow and dry out thawed 1 hour adding the knowledge fluid e 1.2 ml. sterile and 0.9% Iac1 Naya up to 22 sealing mg / ml ation squirrel in substances supernat and in nte ampoule storage (2 ° С) the cloth -28 ° C 2500 Remove -28 ° C 9.6 Introduction Centrifuge Spill into chorion 30 hours Mhz, e 8.2 24 hours, gram 2.1 ml giro marco placent 900 W, ml. 2500 fabrics + 0.5% p- 15 minutes, the bathrooms s 25 sec suckers Mhz, thirty Ra selection of 16.5 ampoules human tsy 900 W, gram formals ml on 3 ml, 27 grams after 30 25 sec sand + on in supernatant lyophile min Double 10 ml. homogenous is defined a little thaw but, 0.9% t on 4 squirrel in vacuum of deleted IAC1 in hours liquids oh 7.7 ml. flow AND dry out suckers 1 hour adding the knowledge tsy e 4.4 ml. sterile 0.9% 1MaC1 Naya up to 22 sealing mg / ml ation squirrel in substances supernat and in nte ampoule storage (2 ° С)

- 16 018848

When administered to the body, a pharmacological composition based on IAA causes a cascade of immunological responses of an individual’s body, leading to controlled damage to the tissue and cells of the same tissue inside the body of an individual, isolated lysis of tissue cells, characterized by the fact that the IAA, which is part of the pharmacological composition, can cause a chain immunological responses from the body’s immune system, in which antibodies are formed to the administered pharmacological composition, which, being antibodies against the introduced antigen are at the same time highly specific autoantibodies to the tissue from which IAB is obtained, or of which IAB is a marker, which is part of the pharmacological composition. A pharmacological composition containing a substance according to claim 1 exhibits properties that lead to sequential and isolated tissue lysis, under the action of specific antibodies formed, which are generated in the body in response to its administration. These properties of the claimed substances can be used for treatment as an antitumor agent in cancer pathology, when removal of tumor tissue from the body is required. A pharmacological composition containing an IAA is capable of marking and damaging tissue, which, before the substance was introduced, grew and developed without control from the body’s immune system, was intact, i.e. did not participate in immune reactions, had uncontrolled growth, did not lyse, did not reject and did not utilize the body on its own and could not be detected by methods of immune laboratory control. The same substance used for diagnostic purposes is able to detect such tissue in the body (to be a tumor marker) and at the same time to be a substance that, under laboratory conditions, is capable of assessing the extent of damage to the same tissue, i.e. can be used to control the level of lytic processes occurring in the body, allowing to assess the degree of tissue rejection.

A method for diagnosing the state of health of the tissue (tissues) of an internal organ of an individual is based on the quantitative determination of autoantibodies produced by the body to the studied tissue of the examined organ or organs. The method includes examining a sample of a biological fluid sample of an individual and determining the amount of antibodies (autoantibodies) formed in a biological fluid after contact with an AAB, to the studied tissue of an internal organ (tissues of internal organs) (anti-tissue antibodies (autoantibodies)) according to the amount of interactions between autoantibodies found in the sample biological fluid of the individual and iai, which is a marker of tissue. The health status of the tissue (tissues) of the internal organs is determined by comparing the digital values (levels) detected in the sample of anti-tissue antibodies (autoantibodies) with the proposed scale of the level of health or tissue damage.

For diagnostic purposes, qualitative and quantitative reactions can be used. Moreover, when a quantitative reaction (enzyme immunoassay) is performed, the maximum possible number of interactions of antigen-antibody, autoantigen (autoantibody) in a controlled laboratory reaction, to complete extraction of antibodies (autoantibodies) from a sample of biological fluid is detected, and the absolute level of autoantibody antibodies in the sample is determined.

When a high-quality immune response is performed, a minimal (at least one) antigen-antibody interaction (autoantigen-autoantibody) is recorded at the maximum dilution (titer) of a sample of a biological sample (in liquid media, for example, RIGA) or a reaction on a solid carrier when the interaction occurred antigen-antibody (autoantigen-autoantibody) can be detected on a solid carrier, for example, by a color change indicating the reaction has occurred.

A qualitative response to the presence (absence) of detectable antibodies (autoantibodies) is taken into account as follows: if at least one antigen-antibody interaction occurs in the original sample or in the resulting dilution (titer) of the sample, which can be registered during laboratory testing, it is considered that the sample has these antibodies (autoantibodies) and the reaction is positive. In the case when the obtained dilution (titer) of the sample does not occur at least one interaction of antigen-antibody, which is not registered during the laboratory study, it is considered that the sample does not have these antibodies (autoantibodies) and the reaction is negative.

Quantitative reaction to the presence (absence) of detectable antibodies (autoantibodies) is carried out in reactions that reveal the total amount (level) of antibodies, autoantibodies, can be evaluated and taken into account in absolute values, and directly in the original sample of biological fluid without dilution, in physical values (µg / ml, µg / l, etc.).

Qualitative reactions can also be quantified. Then the last positive qualitative reaction of the antigen-antibody (autoantigen-autoantibody), which occurred at the maximum dilution of the initial sample, should be taken into account.

In cases where the work is performed with dilutions (titers) of the original sample, the final positive qualitative reaction that takes place at the maximum dilution (titer) of the original sample is taken into account. Then the positive reaction that occurred at this dilution (titer) should be attributed to the set of the scale in which this reaction was revealed (occurred). The scale is given below in table. 2

The coincidence of the obtained indicators in the response of the sample with one of the values of the sets of scale scales

- 17 018848 coincides with this set, and the number of detected autoantibodies corresponding to the level of the detected set (example No. 1 is given below). The resulting titer determines the quantitative content (level) of antibody-autoantibodies in a diluted sample and may indicate the initial level of detectable antibodies-autoantibodies in the undiluted sample. After obtaining the result and comparing it with the scale of tissue health levels, a conclusion is given on the initial state of health and the degree of tissue damage.

The scale of tissue health levels is a scale that has a digital ranking (gradation of dilution levels (titers)) from zero (sample is not diluted) to the maximum dilution level (up to 1: 2048), the sample is diluted 2048 times and has units of measure corresponding to and multiple degrees of breeding. The coincidence of the titer of the reaction with the boundaries of the titers of the scale is considered to be a coincidence of sets of dilutions (titers) of the original sample. Inside the scale, sets of digital values of dilutions, corresponding to the degree of dilution of the original sample under study, are highlighted. If, when taking into account a qualitative reaction, the absence of an antigen-antibody reaction (autoantigen autoantibody) in the dilution under study is detected, the result is considered negative, it is concluded that there is no autoantibody antibody in this dilution.

The sets of dilutions of the original sample are determined by the border numbers inside the scale. The minimum quantitative indicators of the interactions of antigen-antibody, autoantigen-autoantibody, belong to a set of 4-fold dilutions of the sample to a titer of 1: 4 and make up the set [0-1: 4] (not a diluted sample, diluting the sample 2-3-4 times).

Sequences of sets of dilution values in the scale determine the digital indicators identified in immune reactions and which may be associated with or associated with dilutions (titers) of a sample of a biological fluid. These sets of values belong to the set of dilutions of the original sample. The sets of values are within the limits of dilution (titers) of the test sample and represent the five main sets of dilutions of the original sample of the test liquid, with which you can identify the level of antibodies-autoantibodies and determine the state of the tissue under study. These sets are presented below as a scale in the table. 3

Table 3

The scale of the sets of dilutions of the original sample, the levels of the detected antibodies, autoantibodies and the state of the tissue

Dilution rate or sample titer [1: 4-1: 8] [1: 16-1: 32] [1: 64-1: 128] [1: 256-1: 512] [1: 1024-1: 2048] The maximum level of antibody antibodies in µg / ml 12.5 25 50 100 200 Antibody antibody level normal above normal tall excessive critical Fabric condition tissue is healthy minor tissue damage tissue damage typical of chronic inflammation tissue damage typical of acute inflammation tissue damage typical of lysis and / or tissue rejection

Thus, five basic sets of dilutions of the original substance are identified within the scale, which determine the levels of detectable antibodies and autoantibodies and determine the state of the tissue under study. The results obtained are compared with the scale, determining the set to which they belong, and identifying the desired set, determine the resulting value of the result, make a conclusion about the level of antibody-autoantibodies to the tissue in the sample under study. Sets determine detectable levels of antibodies-autoantibodies. After determining the level of autoantibodies in the sample, it is possible to make a conclusion about the state of health or the degree of damage to the individual's tissue.

At the level of the identified autoantibodies belonging to the set [1: 4-1: 8], they speak about the healthy state of the tissue; at the level of identified autoantibodies belonging to the set [1: 16-1: 32], the result indicates the level of antibodies-autoantibodies above the norm, may indicate a chronic pathological process without exacerbation or a recent adverse effect on the tissue. The state of tissue health in this case can be interpreted as a variant of the individual norm; at the level of identified autoantibodies belonging to the set [1: 64-1: 128], the result indicates an exacerbation of the chronic pathological process of the tissue or the beginning process of acute tissue damage, a direct cytotoxic effect of a foreign agent on the tissue or a combination of these factors. The state of health of the tissue should be treated as damage and pathology; at the level of revealed

- 18 018848 leftover autoantibodies belonging to the set [1: 256-1: 512], the result indicates a severe acute pathological process occurring in the internal organ and severe damage to the tissue of this organ, the state of health of the tissue is interpreted as a disease, tissue treatment is required; at the level of detectable autoantibodies belonging to the set [1: 1024-1: 2948], the result indicates the beginning of full or partial lysis of the tissue, its death and rejection. Acutely arisen and (or) maintained level of autoantibodies belonging to this set, as a rule, is not compatible with the life of the whole organism. The state of health of the tissue of an organ and the body should be regarded as critical.

The coincidence of the obtained indices in the reaction of the dilutions of the sample with one of the values of the scale sets is considered to coincide with this set, and the number of autoantibodies detected corresponds to the level of the set detected.

The absence of a result in a qualitative reaction, or the discovery of the results of the reaction in biological fluid equal to zero, indicates tissue organ and (or) species asingnost of the sample and the marker used, the reaction is considered negative and indicates the absence of detectable antibodies (autoantibodies) in the sample under study.

If the result is negative when carrying out a qualitative reaction in an undiluted sample, the level of antibodies-autoantibodies is not determined, then the result of the reaction is considered to be zero, it is considered that there are no antibodies-autoantibodies in the sample of biological fluid. In this case, it is believed that the tissue of the individual:

option 1) has almost no damage, and the process of cell formation prevails over their destruction;

option 2) the tissue has partially or completely changed its antigenic structure, which may be in cases of ΝΕΟ-tissue formation and cancer is underway;

option 3) in the samples of biological fluids of an individual there are substances that block the formation of anti-tissue antibodies-autoantibodies (there was taking or administering drugs that block or reduce the reactions of formation of antibodies-autoantibodies or transplant rejection).

In any of the three options in the body of an individual, a state of tissue arises, named by the author of a tissue autoimmune turn, the state of tissue health in this case requires an in-depth clinical and morphological diagnosis (biopsy, in-depth hardware examination). After obtaining the results of in-depth diagnostics, a definition of the state of health or disease of the tissue of an internal, transplantable, or transient organ can be given.

In the study of a sample of a biological fluid of an individual, in the analysis of which several markers were used simultaneously, where several IABs obtained from different tissues were used as a marker, the result of the interaction with each of the applied IABs obtained from different tissues and the biological fluid of the sample should be evaluated. The result of the interaction for each substance is taken into account separately, and the conclusion about the state of health of each tissue is carried out separately. The totality of the results in the sample and the result obtained for each substance determines the health profile of the tissues in the sample. According to the result of the test, a conclusion can be made about the general state of health of not only the tissue, organ, but the organism of the individual (in case a sufficient amount of AVI is used). The conclusion about the state of health of the individual is made on the basis of the results obtained and the proposed scale, but to each body.

In the study of a sample of a biological fluid sample of an individual, the analysis of which used several markers at the same time, where several IABs obtained from different tissues were used as a marker in the following cases: 1) revealing high titers of antibody levels of autoantibodies in the entire biological fluid sample (titer 1 : 64 and above for each of the analyzed substances); 2) when obtaining the results of the samples, where the variants of the substances according to claim 1 were used in the series and the results for each of them (all values of the titer levels are high or all low), consider that there are a number of biologically and (or) immunoactive substances in the body, leading to disruption of immunological tolerance and (or) hypersensitization of the body with substances capable of leading to dysfunction of the immune system (tissue condition in allergic diseases, a number of autoimmune diseases (autoimmune thyroiditis, etc.), treatment with pharmacological positions containing high doses of the drug substance affecting immune reactions (glucocorticoid hormones, immunosuppressants, cytostatic substances, etc.).

Quantitative determination of the level of antibodies (autoantibodies) is carried out using an enzyme immunoassay reaction, the result is taken into account in the received absolute numerical expression or conditionally accepted units.

Scale values may differ and take absolute numerical expressions in different units of measurement if different options, techniques or devices are used for immune responses. Then digital, quantitative values detected by another method in immune reactions should be compared with the quantitative value of antibody-autoantibodies in the sample dilution titer by the scale of sample dilution levels, and then interpreted.

Qualitative highly sensitive reaction, which allows to carry out the detection using

- 19 018848 substances according to claim 1; the minimum levels of specific antibodies (autoantibodies) to tissues in samples of biological fluids are enzyme-linked immunosorbent assay. The result is considered as positive or negative, and quantitative recalculations are carried out using the proposed scale. In solid-phase immunoassay techniques, which allow to detect the antigen-antibody reactions (autoantigen-autoantibody) that occur, the variants of the substance according to claim 1, obtained by the method of claim 2, can be used. The use of solid carriers for the realization of the reaction can significantly simplify the analysis. It is possible to use the following types of carriers: 1) tubes (for example, glass, polystyrene, polypropylene); 2) granules (for example, activated polystyrene); 3) fine particles (for example, polyacrylamide, cellulose, sepharose); 4) microplates or strips (for example, polystyrene); 5) dipsticks, cards, rods (for example, polystyrene); 6) electrodes (for example, platinum, carbon); 7) films or fibers (for example, glass, quartz). If the system includes an electrode, then it can be called a bio or immunosensor (biochip). An example implementation of immunodOT analysis on a nitrocellulose membrane is shown below.

The claimed IAA can be used for individualized (personalized) treatment of tissue diseases of an individual, which can be characterized by the uncontrolled growth of tissue cells (including tumor tissue of a malignant or benign nature), which require cessation of the growth process and the formation of cells of the same tissue from which Iav; requiring complete or partial lysis and removal of tissue or its cells from the body, including sensitizing the body with an increasing titer of the pharmacological composition in order to cause an organism’s immune system to respond by forming anti-tissue antibodies and antibodies containing antibodies obtained from the same individual and parenterally administered to the same individual by unchecked, by controlling the same levels of antibody-autoantibody formation in the patient’s biological fluids, observing precautions and pro lactics of undesirable damage to other tissues, aimed at preventing the formation of levels of antibody titers-autoantibodies in biological fluids exceeding the values of the set [1: 256-1: 512] in the laboratory control.

The pathogenetic mechanism of action of the proposed method of treatment consists in the directed activation of the immune (autoimmune) system and the production of highly specific antibodies, which, having antigenically identical (syngeneic) sites (organ-species) and an allogeneic site of the organism of an individual in their structure, in response to the introduction of the pharmacological composition, containing iab, are anti-tissue - autoantibodies. The autoantibodies formed by the body have anti-tissue properties and are capable of isolating and directing the tissue with which the fully syngeneic areas, the epitopes, are encoded in which the organ, species and individual properties of the individual's tissue are encrypted. These autoantibodies cannot be isolated from the body and used to treat another individual of the same biological species, but can be identified in its biological fluids using an AVA devoid of the allogeneic properties of the original tissue of the individual (donor) of the same biological species when in the diagnostic system autoantibodies can be detected only in syngeneic areas of organ and species specificity. These properties of a pharmacological composition containing an IAA are key when conducting a therapeutic effect on a tissue, as they allow treatment and control of its result with the same IAA, which, if introduced into the patient's body, is the main active substance, and is the main component of the diagnostic laboratory system that allows to dose and control the course of the therapeutic process.

This is possible, firstly, because the absence of an allogeneic histocompatibility code in an AAB can cause the formation of antibodies in the organism of another individual of the same species, since his body regards the IAV administered as a direct foreign antigen, against which the antibodies begin to produce antibodies and trigger immune responses. -antibody.

Secondly, the complete similarity (initial organ and species syngeness of the tissue) of antigenic organ and species characteristics (codes) of the tissue from which the AVI is derived and which this AVI possesses requires the body’s immune system to develop characteristics that are strictly identical to the organ and species antigenic tissue set, which are implemented in the production of antibodies, in response to the injected substance.

Thirdly, antibodies that have identical organ and species characteristics of an AVA are at the same time identical to the set of antigenic characteristics of an individual's body tissue. As a result, the antibodies produced are able to act on the same tissue inside the body, which indirectly begins to enter into an immune response under the influence of antibodies, which actually become autoantibodies to the individual's own body tissue.

Fourthly, antibodies-autoantibodies formed in the body are highly specific only to the tissue from which they are derived, they are syngeneic to only one tissue in terms of their organ and species set of antigenic properties. Other organ organ systems (matches by organ code) are excluded in the IIA. Consequently, the response of the immune system, in which the formation of antibodies with a different antigenic structure capable of damaging other tissues of the body, is also excluded. Autoantibodies interact

- 20 018848 precision accurate, and only with the tissue from which they are derived (organ and species syngenia). Damage to other tissues of the body is excluded.

For therapeutic effects on body tissue, the following is necessary:

1) obtaining a sample of the tissue to which the therapeutic effect will be applied, and obtaining an AVI by the claimed method;

2) the creation of a pharmacological composition, which will include IAV - tissue marker, obtained by the claimed method from the patient's body, or IAV, obtained by the claimed method from the tissue of another patient (donor) of the same species, if it is revealed that the antigenic characteristics of the donor tissue and future patient match;

3) activation of the immune and autoimmune systems of the patient's body by administering a pharmacological composition containing an AVA method as is used when administering alien sera (modified Bezredka method AM).

Before treatment, the patient is examined. If the patient’s state of health allows the initiation of treatment (laboratory tests, according to the claimed method, did not reveal an individual’s health disorders or damage to other tissues), then before starting the production of samples and the initial administration of a pharmacological composition containing an IAA, premedication should be carried out (0.05-0 , 1 mg / kg of the body weight of the glucorticoid substance to be injected intramuscularly 30-45 minutes before sample placement, and adjust the intravenous infusion of an isotonic solution of any crystalloid, so that by the beginning of the sample it will be EHO is not less than 0.1 ml / kg body weight).

Puncture of the vein should be accompanied by mandatory blood sampling (at least 3.0 ml) for tests to determine the state of health of substances according to claim 1, the method according to claim 8. The level of antibodies (autoantibodies) to the body tissue of an individual to which treatment is applied may be negative in the primary sample or belong to many values [0-1: 4].

If the final form of the pharmacological composition is not prepared and no other instruction for administration is provided, it is necessary to sterilely take 1 mg of dry, lyophilized IAV obtained by the present method, dilute 1 mg of the substance in 1 ml of 0.9% saline. Take 0.25 ml of the resulting solution and dissolve in 4.75 ml of 0.9% saline solution, obtaining a concentration of 50 μg / ml. Take 0.1 ml of the solution and dilute to a volume of 10 ml, obtaining a concentration of 0.5 μg / ml.

The substance thus diluted in an amount of 0.1 ml is injected intracutaneously into the forearm (upper third of the inner surface) to assess the body's sensitivity to the drug. After 30 minutes, sensitivity is assessed by measuring the diameter of the papule. If the papule diameter at the injection site is no more than 1 cm, then another 0.1 ml of diluted serum is injected subcutaneously. In the absence of reaction, after 30 minutes the dose of the substance is dissolved so that the concentration of the solution does not exceed 0.50 μg in 1.0 ml (for example, 50 μg-1 ml of the substance solution according to claim 1 is dissolved in 100 ml of 0.9% saline ). The prepared pharmacological composition for infusion is slowly (20-40 drops / min, 1-2 ml / min) administered intravenously at the rate of not more than 0.01 µg / kg body weight. During the infusion, the state of the vital functions of the body is monitored (monitored) (breathing, pulse, blood pressure). At the first signs of changes in the number of respiratory movements, the number of heart contractions, changes in blood pressure, anxiety of the patient, discomfort - the substance is stopped and the symptoms are stopped by the administration of glucocorticoid substances, antihistamines. The occurrence of the above symptoms is regarded as an interaction reaction that occurred between the body’s immune system and the substance. The patient should be left under stationary observation for at least 24 hours after the procedure for administering the substance.

According to indications, but not less than 12-24-48 hours after the administration, a biological fluid (blood serum) is analyzed, assessing the state of health of the tissues of internal organs and the presence of damage to the tissue of the individual about which the treatment is being carried out. At the same time, the assessment of the state of tissue health is also carried out by the claimed method. In particular, upon receipt of analyzes of biological fluid samples, in which the levels of autoantibody sets were detected at titers [1: 4-1: 8]; [1: 161: 32], therapeutic effects are considered feasible enough. In this case, the patient's body is monitored and the tissue affected by until the desired result is obtained, repeated sessions of the IAV administration are conducted. If the values of the levels of autoantibodies are revealed [1: 64-1: 128], then consider that the therapeutic effect is highly effective, repeated sessions of the substance administration stop. Monitor the patient. At the level of identified autoantibodies belonging to the set [1: 256-1: 512], the therapeutic effect is considered excessive and take measures to reduce the set of values of autoantibody levels, administering glucocorticoid hormones, desensitizing and antihistamine drugs, carry out infusion therapy with crystalloid solutions at the rate of 15- 20 ml / kg, detoxification methods are applied, up to plasmapheresis, until normalization of the set of autoantibody levels in biological fluids is normalized. Exceeding the levels of autoantibodies belonging to the set [1: 1024-1: 2048] is not permissible and may be accompanied by the death of the whole organism.

If the diameter of the papule 30 minutes after the initial injection of diluted pharmacological

- 21 018848 of a composition containing an IVA intracutaneously more than 1 cm, then the following actions are performed: iv administration of 1-2 mg / kg of glucocorticoid hormone (prednisolone), subcutaneous administration of the diluted pharmacological composition containing the substance according to claim .1, in the amount of 0.2, 0.5, 1.0, 2.0, 5.0 ml, then intravenous infusion is carried out, as described above.

When a reaction appears to a fractional dose of serum, 2 mg / kg of body weight of glucocorticoid hormone (prednisolone) is administered i.v. and a sample of biological fluid is analyzed for the level of autoantibodies.

The developed technology allows to obtain new immunologically active substances that are specific to the typed tissue of almost any organ of a human or animal. They are protein substances, namely immunoglobulins, mainly belonging to class M. It has been found that IAA are specific autoantigens to typeable tissue, characterized by signs of protein structures of class C immunoglobulins (1 M), which are involved in urgent serum reactions, have high individual, organ and species specificity, constantly circulating in the serum and having a quantitative dependence on the processes of vital activity and the state of the tissue studied. The amount of tissue autoantibodies detected in serum can vary, having a linear dependence on the morphological state of the substrate. Moreover, the levels of detectable autoantibodies characterize not only the level of metabolic processes in a tissue or organ, but also reflect the state of their damage under the action of exo and endogenous factors affecting this tissue or organ. An increase in the number of detectable autoantibodies in the serum can consistently characterize the state of health of the studied tissue. The claimed method allows to differentiate the levels of practical health, the state of chronic or latent organ damage, the state of acute organ damage and their involvement in the pathological process, to predict the threat of autoimmune damage and organ tissue rejection. Thus, the study is able to identify the state of the body: a latently current illness, an impending health catastrophe, or the critical state of tissue.

Determination of autoantibodies in serum opens up new perspectives for laboratory diagnosis of the state of organs and tissues of humans and animals, allowing to directly assess the state of health or damage to organs and tissues in the sample under study. The claimed group of technical solutions allows you to create a complete line of diagnostic kits for quantifying the level of autoimmune markers in the serum of tissues of organs of animals and humans by ELISA, to determine the autoimmune status of organs of the whole organism.

The analysis to determine the autoimmune status of tissues and organs will allow to directly judge the state of health, threats and the degree of disease of internal organs, monitor the dynamics of recovery of the whole organism or its individual organs.

This technique is a new addition to the existing diagnostic methods. A distinctive feature of this laboratory method is not an indirect (mediated through waste products and organ functions) assessment of their morphological state, but a direct quantitative determination of autoantibodies produced by the body to the organ or tissue under study.

Circulating autoantibodies in the serum are determined by each organ and have their own quantitative values. The number of detected autoantibodies in the serum to the organs or tissues under study in the body can be considered as a new biological constant characterizing the autoimmune system of the body, with a characteristic and variable characteristic of humans and animals. To confirm the possibility of implementing a group of inventions and achieving the claimed technical result, more than 20 such new biological characteristics were established and used to evaluate the autoimmune profile of organs in the human body and more than 70 indicators that characterize the autoimmune state of the animal's health.

According to the claimed method, a series of 20 immunologically active substances was obtained, which were used to diagnose diseases of the internal organs of humans and animals by direct laboratory monitoring of the state of internal organs that are detected in blood serum. Substances were adapted to conduct an enzyme immunoassay and were able to detect organ dysfunctions in the early stages. In the process of conducting research on the obtained IAA, a method for interpreting the results of laboratory tests was developed and unified.

Below is a description of the experiments performed and the results of experiments confirming the presence of partial (partial) organ and species syngality of an immunoactive substance with a description of their properties.

In the process of research the following assumptions were verified.

If the claimed IAV has fully preserved antigenic characteristics of the original organ and the species code of the original tissue (partial, partial syngeness) from which it was obtained, then the feature of the substance will be the incompleteness of the original antigenic code inherent in the entire tissue from which this substance is derived. This incompleteness will determine the partial syngene. Moreover, iai, obtained by the claimed method, with fully preserved properties of antigenic characteristics

- 22 018848 risk of organ tissue, individual antigenic properties of the same original tissue must be leveled so that it will be able to enter into an immune response antigen-antibody, autoantigen-autoantibody in biological fluids taken from different individuals of the same biological species, only with specific autoantibodies body, without entering into nonspecific reactions that can be triggered by the characteristics of individual differences in samples from different subjects. If a substance is able to affect the activity of the immune and autoimmune systems of the body, it is only through a cascade of immune and autoimmune reactions that will produce antibodies (autoantibodies) in response to the administration of an IAA indirectly, affecting increased levels of anti-autoantibodies on the tissue itself. Antibodies to a substance having a partial organ syngality, which are formed in response to its introduction, will be identical to the tissue of the organ from which this substance is obtained. The production of antibodies by the body's immune system, in response to the administration of an AVI, has strict patterns. Antibodies according to their information code are identical to the tissue of the organ from which the substance was obtained. Antibodies produced by the body are antibodies against the substance. But the substance has a complete antigenic similarity (organ syngennost) with the tissue of the organ from which it is derived. Therefore, the antibodies produced will have the properties of autoantibodies to the tissue of the organ of the body, the tissue of which is similar to the AVI from which it is derived. This means that antibodies produced by the immune system in response to the introduction of the substance will have anti-organ, anti-tissue characteristics and properties. These antiorgan autoantibodies will have an individual body code, which allows them not to interfere with the immune response and not to damage the cells of the tissues of other organs of the individual, but to have only the anti-organ anti-tissue damaging effect.

The object of the study was the IAA, obtained by the claimed method from guinea pig liver tissue.

At the same time, to obtain an IAA, the initial tissue was collected, typed, preserved by freezing, after which the initial tissue was subjected to a multiple process of freezing and thawing. The defrosting of the fabric was carried out by exposure to microwave radiation with the removal of moisture formed in the process of thawing. The fabric was dehydrated by 43% (until a residual moisture content of 57% was reached). The microwave effect was carried out at a frequency of the wavelength of 2450–2500 MHz with a radiation source power of 700–900 W for a time of 20 s, at which water crystallized on the surface of frozen tissue turned into a liquid state. The initial fabric obtained as a result of this treatment was crushed, homogenized, disinfected by adding 0.5% aqueous formalin solution to the tissue homogenate to 5-10% of the total starting material, followed by mixing and exposing the mixture for 4 hours and extracting it according to Grasse, the supernatant was subjected to standardization of protein from 20-25 mg per 1 ml and lyophilized.

The experimental material obtained in this way in the form of an IAA substance was parenterally administered to laboratory animals — guinea pigs and laboratory rats, selected at random, not having syngeneic inbred lines. The experiment involved 27 healthy animals (14 guinea pigs (350-450 g) and 13 laboratory rats (250-400 g)). The experiment was carried out on young (2-3 months) apathogenic animals. The animals were kept in accordance with the rules of the European Convention for the Protection of Vertebrate Animals used for scientific purposes. At the end of the experiment, animals were killed by the method of cervical dislocation in compliance with the Rules of work using experimental animals, approved by order of the Ministry of Health of Russia.

In the group of guinea pigs, a control was determined, a group of 3 animals, one of which was used to collect liver tissue, from which the AVA was obtained by the claimed method. This tissue was also examined by electron microscopy and was taken as an ultrastructural control sample and a standard of intact tissue of the normal liver of an animal, against which experimental samples were compared.

Liver tissue samples were fixed in a 2.3% glutaraldehyde solution in 0.1 M cacodylate buffer for 1 h with refluxing in 1% О§О _four according to S. Raile (1952). After dehydration, the drug was poured into epoxy resins by the method of A. Zrigg (1969). The fabric was contrasted in pieces of a 0.5% solution of uranyl acetate and on the nets according to G. Keupoles (1963). Sections were prepared on a BKB ultratome. The preparations were examined on a NyaeY 12-12 A microscope with an accelerating voltage of 75 kV.

Blood was collected from all animals for the study of serum for the level of anti-organ hepatitis autoantibodies and anti-tissue autoantibodies to the tissues of the heart, lungs, brain and kidneys. Below is a table. 4 averaged values of the initial autoantibody titers to the tissues of the internal organs in the subgroups of experimental animals - the tissues of the heart, lungs, liver and kidneys.

Table 4

Experimental subgroup A heart Lungs Liver Kidney Guinea pigs 1: 4 1: 8 1: 4 1: 4 Rats 1: 4 D.4 1: 4 1: 8

To detect the level of anti-tissue anti-organ autoantibodies in rats, classical serological immunological reactions were carried out, and methods of immune reactions were used, in which positive reactions were detected colorometrically and could be reduced to qualitative or quantitative indicators, depending on the current laboratory task. Reactions were carried out according to the traditional method, in which the AAA obtained by the claimed method was used as a marker. To interpret the values of the titles were used previously given recommendations. It is necessary to make a general conclusion that the level of autoantibody titers to the examined tissues of the internal organs of experimental animals does not go beyond the physiological norm. Internal organs do not have laboratory data that allows them to be regarded as damaged or diseased.

Next, all the animals of the experimental group (11 guinea pigs and 12 rats) were administered the proposed AIA in the initial estimated dose of 0.01 μg. After 4-6 hours, the motor and food activity of all animals decreased, and in the experimental subgroup of rats, disorders appeared 1-1.5 hours earlier. After 12-18 hours, the motor and food activity of the guinea pigs began to recover to its original level. All animals survived the initial dose of the injected substance for at least 48 hours. In the experimental group of rats, motor and food activity did not fully recover after 24 hours, two animals died, 10 animals survived 48 hours.

After 48 h 6, guinea pigs and 7 rats were reintroduced the calculated full dose of the analyte according to the previously developed scheme after premedication with prednisone. 24 hours after repeated administration of the substance (72 hours of the experiment), 2 animals fell in the experimental subgroup of guinea pigs, 4 animals underwent repeated administration of the preparation. Two out of five animals that were injected 72 hours ago, again reduced motor and nutritional activity, three animals that did not undergo repeated administration of the drug, did not show health disorders, decreased motor and nutritional activity in comparison with the control group. After 120 h, 8 animals remained alive in the experimental subgroup of guinea pigs (an animal fell, which received a 1-fold introduction of the substance 100 h ago). Two animals from among those who received repeated administration of the substance had a painful appearance: they did not move and did not feed. They fell within the next 12-16 hours. After 168 hours (7 days), all pigs that received repeated administration of the substance, and one pig that received 1-fold injection of the substance in the first 24 hours of the experiment, fell.

48 hours later, in the experimental subgroup of laboratory rats, 1–2 hours after the administration of the preparation, 3 animals died again, the rest fell 6–8 hours after the repeated administration of the substance. The dead animals were opened and their internal organs were examined. All animals showed signs that characterized the rat's response to a toxic-allergic form of shock (anaphylactic shock). Experimental animals (3 rats) that did not reintroduce the substance also fell within 72-80 hours of the experiment. Thus, in the experimental group of rats, all animals that were injected with the substance died. Repeated administration of the substance dramatically aggravated the condition of the experimental animal and led to its death within a maximum of 6 hours after repeated administration. All animals from the control groups (rat and guinea pigs) and two pigs from the experimental subgroup, which received 1-fold maintenance of the substance on the first day of the experiment, remained alive after 240 hours (after 10 days). Animals from the experimental group of guinea pigs that survived the 1-fold injection of the substance were slaughtered and they had a study of the internal organs. At autopsy, localized areas of liver tissue damage were found in both animals. They had a star-shaped scar tissue with a zone of moderate inflammation. These sections of stromal liver were subjected to conventional and electron microscopy.

FIG. 1-6 presents electron diffraction patterns of the liver tissue ultrastructure of animals in the control sample and in the experimental group.

As the main control tissue sample, one of the typical microelectron diffraction patterns of an intact guinea pig liver tissue is shown. FIG. Figure 1 shows the ultrastructure of guinea pig liver in a control sample with 8000 magnification.

FIG. 1 fat drops are visible (F); lumen of a sinusoid (C); Disse space (D); parenchymal liver cells (P); the core of the lyiocyte (I); mitochondria (M). Liver cells (hepatocytes) make up about 50-60% of the mass of guinea pig liver. They have a polymorphic polygonal shape and a diameter of approximately 30-50 microns. These are one or two nuclear, less often multinucleated cells that divide by mitosis. The lifespan of hepatocytes in experimental animals is about 150 days. The hepatocyte is bordered by a sinusoid and Disse space, with the bile duct and adjacent hepatocytes.

In the control - the intact tissue of the liver of the animal - the hepatic parenchymal hepatic cells are clearly pronounced, have contouring rounded nuclei and good chromatin condensation. The membranes are contoured, structural, well-defined. The nucleus of a star-shaped cell (lipocyte) has an oval shape and a well-structured nuclear membrane. Cytoplasm has well-defined outgrowths and deposits in the form of fatty spots. The cytoplasm is filled with electron-dense granules with non-uniform density. FIG. 1, mitochondria and the structural part of the endothelial cell (sinusoid) are distinguishable.

Were obtained dilutions of the proposed IAV 0.1 μg-10 ml The resulting solution was injected

- 24 018848 to animals at the rate of a full dose of 0.01 µg for every 100 g of body weight of an animal fractionally, subcutaneously and intramuscularly with an interval of 30 min, starting with a dose of 0.01 µg. As a premedication, prednisone 5 mg was used intramuscularly 1 hour before the substance was introduced to all animals without taking into account body weight and the type of animal. The blood of the animal was taken before the substance was administered and then repeated every other day, determining the values of the levels of autoantibodies to the tissue of the liver, heart and lungs. Observation of the animals was carried out in the period of the entire effect of the substance on the animal, after the introduction of the initial dose.

Microscopic examination revealed areas of tissue sclerosis, limited to the zone of inflammation. An in-depth electron microscopic study obtained a picture of the damage that occurs in the tissue upon parenteral administration of the proposed substance, which is not reversible and completely deprives the tissue of typical cells. FIG. 2 shows the electron diffraction pattern of the site of local damage to the liver of a guinea pig, which has arisen in vivo under the influence of an AVA from a tissue of the liver of a guinea pig (magnification x7500).

FIG. 2 shows the result of the damaging effect of the claimed IAA on the identical tissue of the guinea pig liver of another individual. Such tissue changes could occur under the action of only a highly specific agent that has almost complete genetic affinity with the tissue. Moreover, the changes are in vivo and tissue devoid of specific areas (liver parenchyma), is sclerosed. The liver tissue, which survived the damaging effect of the claimed IAV, is a cytoskeleton of endothelial cells of the liver tissue, free of hepatocytes, containing membrane remnants and contents of stellate cells that completely lost fat drops, resembling fibroblast and myofiblast structures, scar tissue of the liver.

Taking into account the fact that the study drug was taken from a live animal that did not have signs of disease, or rather survived injury, it is safe to say that the guinea pig liver tissue under study, with parenteral administration, has an isolated effect on the liver tissue. Moreover, at the opening of the experimental animal, no pronounced changes in other internal organs were detected that would go beyond the accepted norms. This circumstance once again confirms the assertions that the substance under investigation has an isolated effect only on the specific tissue, the one from which it was obtained.

In the course of the experiment, all the corpses of animals of the experimental subgroup of guinea pigs were gradually dissected and pathological and autopsy were performed. The results of the autopsy were not pronounced differences. Animals fell from liver tissue damage. At autopsy, signs of acute liver tissue necrosis, acute dystrophy, hepatitis, biliary cirrhosis, jaundice were detected. All animals lost 10 to 25% of their initial weight.

Below is a typical result and the conclusion of a pathological and pathological-anatomical study of the corpse of an experimental animal - a guinea pig, which fell after the introduction of an AVI.

Single hemorrhages of the mucous membrane and peritoneum were found, there was no effusion in the pleural cavity, moderate swelling of the internal organs, swelling and swelling of the lymph nodes of the portal zone, hemorrhages in the lymph nodes, lemon-colored sclera and mucous membranes. The spleen is enlarged, dense to the touch with dense full-blooded pulp; the color under the capsule is purple-brown, red-brown on the section. The liver is swollen, filled with blood, a mosaic pattern on the background of a general reddish-brown color, there are small limited patches of orange-yellow-white color, the edges are rounded. Fibrinous inflammation of the liver capsule is detected with abundant deposition of fibrin on the surface areas of the liver, which has changed color, in the form of gelatinous, easily separated masses. The liver itself at the same time increased in volume, flabby consistency. The gallbladder with a viscous dark red-brown bile. The kidneys are swollen, the capsule is easily removed, the cortical substance is dark red, and the brain is gray-red. Urinary bladder contains rich yellow urine. The heart has a rounded shape, tight-elastic, red-brown in the cut, blood clots in the cavities. Myocardium is red, valves are not changed, without hemorrhages. The lungs are pink, somewhat full-blooded, with dense areas elevated above the surface, without hemorrhages under the pleura. There is a small amount of mucus in the bronchi. The mucous membrane of the stomach is not changed. The mucosa of the small intestine is covered with a layer of thick mucus. The brain is wet, the convolutions are well contoured, white and gray matter of the brain is differentiated.

Pathoanatomical diagnoses: jaundice; regional hemorrhagic lymphadenitis; acute hepatitis; signs of biliary cirrhosis.

Experimental conclusion: primary acute isolated liver tissue damage, secondary changes in the tissues of internal organs.

Laboratory data for assessing the levels of autoantibodies to the tissues of the internal organs in a sample of the blood serum of a guinea pig that fell 70 hours after the start of the experiment and received 2-fold administration of the test substance are presented in Table. five.

- 25 018848

Table 5

Experimental subgroup A heart Lungs Liver Kidney Guinea pig fallen after 70 hours of experiment 1:64 1:64 1: 1052 1: 128

From tab. 5 shows that the autoantibody titers to the heart and lung tissue are higher than normal, as well as the values of autoantibody titers to the kidney tissue, which may indicate acute damage or disease of the internal organs. But above all, the titer of autoantibodies to liver tissue attracts attention, which indicates its direct acute and severe damage and lysis. It should be noted that the test substance was used as a marker of the laboratory agent in the immune response.

The following are the results of staged electron microscopy of liver tissue exposed to the test substance. Liver tissue preparations for electron microscopy were prepared according to the procedure described above. Studies of liver tissue were carried out in stages in dead animals.

FIG. 3 shows the electron diffraction pattern of the liver tissue of a guinea pig, which has undergone a 2-fold administration of the test substance, and which has fallen after 100 hours of the start of the experiment. An increase of x8000.

We present the data from a study of the liver tissue of an experimental animal (guinea pig) that underwent and fell 100 hours after the start of the experiment (liver tissue at 8,000-fold magnification with an electron microscope).

FIG. 3 shows the result of the damaging action of the agent (the test substance according to claim 1). Visible balloon dystrophy of liver cells, lysis and damage to cell membranes, destruction and death of the nuclear apparatus, cytoplasm and cellular elements, with full or partial release of lysis products into the interstitial tissue and the internal environment of the organ and organism are visible. Declare IAV has a specific damaging effect on the liver tissue. Such changes can lead to subsequent tissue destruction, disruption of cellular relations within the tissue, with the possibility of hardening the damaged areas.

On the electron diffraction pattern (Fig. 4, magnification x5600) of another drug, damage occurring in the liver tissue of the guinea pig, which fell after 72 hours at the start of the experiment after repeated administration of the test substance, is visible. FIG. 4 shows the accumulation of nuclear chromatin and areas of rupture, damage to cell membranes.

On the next preparation, obtained from the liver tissue of a guinea pig, which also fell after 72 hours of the start of the experiment after re-injection of the test substance (Fig. 5 is an electron diffraction pattern with 8200-fold magnification), filament accumulation zones are visible, mosaically ordered, characterizing cytoplasmic condensation of protein structures , Collagenization of Disse space, scattered fragments of the filaments and the chromatin nucleus located outside the cell membrane, fat drops and a pattern of balloon degeneration of liver cells, a conglomer at-shadow cells, dystrophically altered parenchyma cells. Ultrastructural analysis of cell shadows showed that the first sign of destruction was the destruction of the nuclear membrane, then chromatin lysis occurred with still relatively intact cytoplasm, transformation of the cytoplasm and its consequent dystrophy, with the formation of a picture of balloon hepatocyte dystrophy. This staging of liver cell damage is visible in FIG. five.

Thus, hepatocytes of various sizes are visualized, their polymorphic and heart-shaped forms are absent. All hepatocytes have a round-oval shape, which may indicate an increase in intracellular pressure in response to damage to the intracellular structure and cellular elements. These circumstances may determine the varying degree of chromatin concentration inside the cell. Visible damage to cell membranes. The membranes have a different thickness, which also indicates their participation in the process of cell damage. Some hepatocytes have signs of balloon degeneration, and in a number of cells lysis of their cytoplasm is seen. The nuclei of stellate cells are deformed, their membranes are heterogeneous and damaged, the cytoplasm is heterogeneous. Drops of fat and chromatin nucleus in the interstitial tissue. The absence of nuclei within the vast majority of heptocytes. The picture shows the phasing of tissue damage, which in turn suggests that it is possible to carry out a phased and metered effect on tissue lysis processes, the ability to evaluate and effectively influence the strength and magnitude of damage inflicted.

FIG. 6 shows the electron diffraction pattern of the ultrastructure of guinea pig liver tissue, which fell 132 hours after the start of the experiment and received a 2-fold dose of the test substance. Electron diffraction pattern is made at 8000 magnification. FIG. 6 shows the seal of the interstitial tissue, Disse space, the seal of intracellular structures and cytoplasm, hepatocyte hardening. The rupture of the membrane of the lipotocyte and the pronounced areas of its thickening, along with the condensation and compaction of the internal structure of the star cell, can be characterized as the beginning of the process of sclerosis of tissue, subjected to dystrophy and necrosis. This may indicate intravital processes inside the organ leading to the restoration of tissue viability after injury.

- 26 018848 waiting, locality of damage.

Thus, it can be concluded that administration of the inventive substance being tested, obtained from guinea pig liver tissue, leads to isolated damage to the liver tissue parenchyma of an experimental guinea pig animal, without damaging the other internal tissues of the animal. Damage caused by a substance is isolated but irreversible, it can be adjusted according to the strength and volume of damage. An experimental animal is able to tolerate the administration of a substance.

The experimental subgroup, which united laboratory rats, suffered the heavier administration of the test substance. Among the animals of this subgroup, there were no surviving individuals who had undergone a 2-fold injection of the substance. All animals fell without surviving the 50 h experiment. After 120 h of the experiment in the experimental subgroup of laboratory rats, only one animal remained alive (control). The control animal was observed for up to 10 days and had no signs of disease or lifestyle changes.

Below is a typical result and the conclusion of the pathological and pathological-anatomical study of the corpses of experimental animals - rats.

Hemorrhages of mucous membranes and peritoneum were found, a red transudate in the pleural cavity, marked edema of the internal organs, swelling and swelling of the lymph nodes that have a smooth structure and gray-pink or gray-yellow color, hemorrhages in the lymph nodes, lemon color sclera and mucous. The spleen is hyperplastic: dense, with round edges, a strained capsule, with a bulging dense pulp; the color under the capsule is purple, red-brown on the cut. The liver is swollen, dense, fragile, orange-brown in color, the edges are rounded. The gallbladder is filled with a viscous dark red-brown bile. The kidneys are flabby, swollen, the capsule is easily removed, the edges of the incision do not converge, the cortical substance is dark red, and the brain is gray-red. The bladder contains urine of a thick red with a brown tint of color, which is not clarified on standing, that is, is colored with hemoglobin. The mucous membrane of the bladder is swollen, with hemorrhages. The heart has a rounded shape and loose pale red clots in the cavities. Myocardium is gray-pink with yellowish areas, dull, easily crushed by the handle of a scalpel; under the epicardium - hemorrhages. The lungs are pink, crepitating, with dense patches of red and white with a hemorrhagic rim elevated above the surface; with diffuse hemorrhages under the pulmonary pleura; with small foci of emphysema. The bronchi contain thick pink foam. The mucous membrane of the stomach is littered with black-colored hemorrhages. The mucosa of the small intestine is thickened, hyperemic, covered with a layer of thick yellowish mucus. Tin brain slightly swollen, wet, gyrus smoothed.

Pathoanatomical diagnoses: hemorrhagic diathesis, spleen hyperplasia, serous-hemorrhagic lymphadenitis, acute hepatitis, jaundice, anemia, protein nephrosis, hemoglobinuria, acute alterative myocarditis, hemorrhagic pneumonia, catarrhal enteritis, brain disease.

Experimental conclusion: acute toxic-allergic damage to the tissues of internal organs, with a primary lesion of the lungs, liver, kidneys.

Laboratory data assessing the levels of autoantibodies (autoantibody titer values) to the tissues of internal organs in a sample of blood serum of a laboratory rat male posthumously collected 50 h after the start of the experiment, and fallen 2 h after repeated administration of the inventive substance are presented in Table. 6

Table 6

Experimental subgroup A heart Lungs Liver Kidney Guinea pig fallen after 70 hours of experiment 1: 128 1: 256 1: 512 1: 512

The analysis of autoantibody titers to the tissues of the internal organs of an experimental rat allows to make a conclusion about an acute severe general lesion of the tissues of internal organs that are not compatible with life. Electron microscopy of rat liver tissue was not performed.

The totality of the obtained data from the survey, observation and laboratory studies allow us to draw the following conclusion.

The test substance has a severe overall effect on the body of an experimental animal of another species, leading to life-incompatible consequences and damage to internal organs. The damaging effect of the substance is based on a typical anaphylactic reaction, leading to shock and death of the animal, which develops in response to its administration, as a high-birth antigen. The predominant site of damage is the life support organs of the experimental animal. The existing damage to the liver of an experimental animal cannot convincingly indicate locally inflicted damage only to this tissue.

- 27 018848

Studies and experiments show that:

the claimed IAA is devoid of the allogeneic properties of the original tissue of the body, which has a highly specific effect on the body’s immune (autoimmune) system, manifested in the fact that the body’s immune system is capable of producing antibodies in response to its introduction;

The claimed IAA has at least one antigenic (partially or partially syngeneic, organosingen) site in its composition, which causes an isolated highly specific response of the immune system (autoimmune) of the organism of the same species, manifested in the lysis of the similar tissue inside a living organism, in response on its parenteral administration (immunization);

the claimed IAA causes a highly specific response of the immune (autoimmune) system in the organisms of individuals of other species, manifested in the anaphylactic shock reaction of a living organism in response to its parenteral administration (immunization), leading to death of the organism as a reaction to the introduction of a foreign antigen;

The claimed IAA has at least one antigenic (syngeneic) site in its composition, which is able to detect highly specific antibodies (autoantibodies) against a tissue of a living organism circulating in the body's internal biological fluids (including serum) of the same type, in laboratory immune responses, forming antigen-antibody complexes;

the resulting isolated lysis of a similar tissue inside a living organism in response to parenteral administration (immunization) of the claimed substance is due to the increasing titer of partially syngeneic anti-tissue, anti-organ antibodies (autoantibodies).

Thus, the claimed substance is characterized by the following combination of properties: the ability to induce an immune response, to cause isolated lysis of typical cells of the tissue to which it belongs and (or) from which it is obtained without causing damage to non-typical tissue (stroma of the organ tissue) and other organs in a living body. The claimed combination of properties allows you to use the claimed IAA as a marker in the implementation of laboratory control for:

the level of tissue damage by identifying the level of autoantibodies produced by the body to the substance and being anti-tissue autoantibodies of the body to the same tissue;

the state of health of an individual by detecting damage to the tissues of internal organs by laboratory determination of the level of anti-tissue autoantibodies produced by the body during illness and in health.

Example 1. Identification of a marker of human liver tissue using enzyme immunoassay.

Detection of antibodies to the substance marker of human liver tissue in serum using enzyme immunoassay.

The presence of specific autoantibodies (class IgM) to human liver tissue in human serum was tested and established using enzyme immunoassay. Conducted a series of experiments with a substance derived from human liver tissue. FIG. 11 shows the curves of the obtained data. ELISA revealed specific autoantibodies to human liver tissue of the IgG class.

Example 2. The use of substances according to claim 1, obtained by the method according to claim 2, for estimating the level of antibody antibodies to the tissues of organs in реакцииΓ реакции (reaction of indirect hemocoagulation).

The reaction of indirect hemocoagulation is carried out according to the classical method. For sensitization of formalized sheep erythrocytes, substances are selected by π. 1, obtained by the method according to claim 2. In the given example, substances obtained from tissues of the heart, liver, kidneys, stomach, vascular tissue of the placenta and chorion were used. The result of the interaction of substances according to claim 1, obtained by the method according to claim 2, and the anti-organ antibodies-autoantibodies in the dilutions of the patient's blood serum was evaluated visually 90 minutes after the reaction was set. The result was recorded in the protocol. Based on the following provisions:

+ - positive result, assessed visually by the characteristic shape of an umbrella;

± - boundary result, not an umbrella or a ring;

And - a negative result, is estimated visually by the characteristic form of the ring.

For the study, the patient's blood serum was selected, after the study a titer of autoantibodies to the tissues of the studied organs was detected (Fig. 12).

The result is summarized by the protocol, which is given in table. 7

-28018848

Table 7

Protocol of levels of antibodies-autoantibodies to tissues in the reaction of rnga

Patient O., age 36 years old, diagnosis: pregnancy 40 weeks, moderate gestosis

Substances under item 1, The titer of autoantibodies obtained by the method of p.2 used as Norm Chronic Acute process phase organ tissue marker: process or the threat of rejection 1: 4 1: 8 1:16 1:32 1:64 1: 128 1: 256 1: 512 Goal. brain (white tissue The study was not conducted substances) Thyroid Thymus The study was not conducted Heart (myocardial tissue) + + ± А№1 Lungs (alveolar tissue) The study was not conducted Liver (tissue + + ± parenchyma) E number 5 Stomach (smooth + + ± muscle tissue) C number 3 Kidney (parenchyma tissue) + + + ± В№2 Skin (Epidermis) The study was not conducted Testicles Tezbz (husband.) The study was not conducted Placenta (vessels) P No. 4 + + + + + + + + Placenta (shell) The study was not conducted Placenta (chorion) E№6 + + + + + + + +

The protocol contains high titers of antibodies — autoantibodies to placental tissue (chorion and vessels). The placenta is a transient organ that is rejected during childbirth. The examinee has 40 weeks gestation. Conclusion: Mature placenta, is its rejection. The patient entered childbirth. This patient began labor activity 6 hours after the analysis.

Example 3. HPLC analysis of substances according to claim 1, obtained by the method according to claim 2, from human liver and kidney tissue.

Studies have been conducted on prepared lyophilized samples of substances according to claim 1, obtained by the method according to claim 2, from the tissues of various organs. Samples were dissolved in 25 mM Tp g-HC1 buffer (pH 7.2) to a concentration of 0.5 mg / ml, and further desalted on an RE-10 column (containing Zeribekh C-25). HPLC analysis was performed using an 8taPG1pe 5000 chromatographic system (Kpieeg) on a T8K Vyuzzh! ^ Column (5x50 mm), under the following conditions: buffer — 25 tM Tp g-HC1 (pH 7.2); linear gradient -0 ^ 1 M №С1 within 60 min; speed - 0.5 ml / min; photometric detection at 280 nm; the volume of the applied sample is 1 ml.

In the course of the study, it was found that the characteristic for all samples of substances according to claim 1, obtained by the method according to claim 2, is the presence of 2-3 dominant fractions of alkaline components, weakly interacting with the anionic column at the pH value used, as well as different acid charge major components eluted in the middle and at the end of chromatography with stable peak ranking numbers in the range of 17 ± 2.5; 33.5 ± 2.5 and 52.5 ± 2.5 tAi (Fig. 7 and 8; kidneys and liver, HPLC charts).

Example 4. Electrophoretic analysis of substances according to claim 1, obtained by the method according to claim 6.

For electrophoresis, the samples of substances according to claim 1, obtained by the method of claim 6, were taken from rabbit stomach tissue, human stomach tissue, human liver tissue, human underwear tissue, human brain tissue, human testicular tissue. Samples are arranged for electrophoresis in the order given above, from left to right, and to the right of FIG. 8 shows an atomic mass scale (kDa) rating scale. Electrophoretic analysis of samples of substances was carried out by a standard method in a 12.5% polyacrylamide gel in the presence of sodium dodecyl sulfate. At the end of electrophoresis, the gels were stained with silver nitrate. The obtained analysis revealed the presence of at least two major bands with Mm of about 10 and 60 kDa characteristic of all samples (Fig. 9).

- 29 018848

Example 5. ELISA on a solid carrier, immunoDOT analysis.

Sequential double dilutions of the analyzed substances (taken in the initial concentration of 100 μg / ml) were applied to the nitrocellulose membrane in the center of the outlined square with a side of 5 mm and kept the samples in a dry-heat oven for 5 minutes at 40 ° C. After blocking the nonspecific sorption sites for 30 minutes at room temperature in 1% BSA solution (bovine serum albumin), the samples were placed in an envelope of RagaIl film and incubated in the patient's serum, phosphorus-buffered solution (1) 1) containing 0.01% BSA and 0.02% Tween-20 and incubated overnight at 4 ° C. After washing with PBS containing 0.02% Tween-20 (3 times 15 minutes each time), the samples were placed in a solution of peroxidase-labeled sheep antibodies to 1dC or 1dM human containing 0.01% BSA and 0.02% Tween-20 (final the concentration of these antibodies was 2 μg / ml). After 2-fold laundering of PBS (containing 0.02% Tween-20), the membranes with applied samples were placed in a substrate solution to evaluate peroxidase activity, which is 0.03% diaminobenzidine and 0.02% hydrogen peroxide in Tris-HC1 buffer (pH 8, 2). Evaluated the results.

It was found that even a short-term immobilization of samples on the solid phase allows to obtain reliable results for the detection of anti-organ, anti-tissue antibodies in healthy people, to give conclusions about the state of health, the examined tissues, to predict the immediate health results.

We give examples of the result of immunoDOT analysis in healthy volunteers I. and Sh. (Fig. 10) conducted on a solid carrier with anti 1dM and anti 1dS. at a dilution of substances according to claim 1, obtained by the method according to claim 6, from 200 μg / ml to 12.5 μg / ml, from 5 tissues (stomach of the rabbit, liver, kidneys, testicles and human brain).

The claimed technology can also be used to prevent complications during vaccination.

The use of the proposed method of diagnosis for the selection of patients during mass vaccination allows you to identify patients with hidden pathology who have no clinical signs of existing diseases. Most vaccines are intended to be administered to healthy people or patients for whom the course of the disease or condition of the vaccine will not have a negative impact. The situation when the complications after vaccination outweigh the positive effects is possible, but only if the body is not able to transfer the vaccine (there are hidden health disorders or diseases of the internal organs, which at the time of vaccination have no clinical manifestations and cannot be detected by conventional laboratory methods diagnostics). Therefore, vaccination has a risk for an externally healthy organism, and there is a possibility that immunization will not give the desired result when risk factors can significantly aggravate latent diseases in the human body, or even lead to death. Currently, relative (temporary) contraindications for mass vaccinations have been introduced for people suffering from any acute illness or having exacerbations of chronic diseases. Vaccines are not administered to such people in order to ensure their maximum safety and reduce the risk of developing complications under the vaccine action, since their disease can be significantly exacerbated by the negative effect of these drugs and lead to serious complications. However, there are currently no reliable ways to detect the exacerbation of chronic diseases or the latent period (prodrome) of an acute disease. The use of substances according to claim 1, obtained by the method according to claim 6, can be used to identify acute diseases and to identify the expected phases of exacerbation of chronic diseases in the latent phase of the process (the period of imaginary clinical well-being). For these purposes, the health of the prospective patient is diagnosed and a laboratory test is made at once with several substances according to claim 1, obtained from the tissues of various organs. Determine the level of protivotkanevyh, antiorganic antibodies, autoantibodies to the main (vital) organs of the individual (heart, vessels, liver, kidneys, lungs, stomach, intestines, thymus, thyroid and pancreas) by one of the methods described in the invention. The result obtained is analyzed, and if the detected levels of the detected autoantibody antibodies (on a scale) do not exceed the dilution titers of 1:32, and the absolute values of the parameters are determined lower than 25 μg / ml for all tissues examined, it is concluded that vaccination is possible. patient without the risk of severe complications. If, at least in one laboratory line characterizing the level of antibody-autoantibodies of the tissues listed above, dilution titers exceeding the above figures are detected, then such a patient should not be vaccinated because of the risk of developing complications.

The mechanism of early diagnosis is based on the ability to detect levels of specific 1dM organ tissues in a patient's biological samples using substances according to claim 1, obtained by the method according to claim 6, which are laboratory precursors of future exacerbations during the development of severe disease and are detected in blood for 48-120 h before its clinical manifestation.

As an example of the early detection of high levels of antibody-autoantibodies in a patient with normal laboratory clinical and biochemical parameters in biological samples

- 30 018848 fluids, here is a clinical case.

Clinical observation. Case history SarNIITO.

(Permission to publish medical history data is coordinated with the attending physician, the head of the medical institution and the patient’s relatives).

Patient D., 38 years old, diagnosed with Arnold Chiari syndrome. Spinal cord hypoplasia.

Medical history data: ill for 11 years when, against the background of complete health, muscular weakness of the right upper limb appeared and, after a long examination, the above diagnosis was established. Symptoms increased despite treatment. Consistently joined muscle weakness in all limbs, muscles of the body, respiratory muscles. Operated in 2006 on the spine. There was a short remission. The last exacerbation occurred 14 days ago, the patient was hospitalized in the hospital. Satisfactory condition. Complaints of pronounced weakness, subjective feeling of lack of air due to lack of strength for breathing and a restriction in the ability to take a deep breath, weakness in the limbs and heavy fatigue.

Objectively: 1 ° = 36.0 ° С. Consciousness is clear, oriented, adequate. Overall condition is satisfactory. Normostenic body type, normal nutrition. The skin is clean, flesh-colored, normal moisture. Visible mucous membranes of normal moisture, pale pink. The bone marrow system, the mammary glands are not changed. Peripheral lymph nodes, thyroid gland not enlarged. The chest is the correct form, both halves simultaneously take part in the act of breathing.

Forced breath is performed, in his participation takes the abdominal wall and the diaphragm. No shortness of breath. NPV - 18-20 in 1 min. Perkutorno over the lungs a clear pulmonary sound, no wheezing, no crepitus; the lower boundaries of the lungs are not changed. Auscultatory vesicular breathing. Borders of the heart are not changed by percussion. Tones sonorous, rhythmic. HELL - 120/70 mm Hg Pulse - 72 beats. in 1 min, synchronous, rhythmic, satisfactory filling and tension. The tongue is coated with white bloom, wet. The pharynx is not hyperemic. Pharyngeal tonsils are not enlarged. The abdomen is of the usual form, palpation is soft, painless. The liver is not enlarged. The spleen is not palpable. Physiological functions are not affected. Symptom tapping on the lumbar region is negative on both sides. Decreased tendon reflexes. Symptoms of irritation of the meninges are absent. There are no gross neurological symptoms.

Laboratory data:

Total blood test: er. - 4.96x10 ¹² / l, Hb - 155 g / l, CPU - 0.94, ESR - 17 mm / h; Leykots. - 5.7x10 ⁹ / l, eos. - 2%, and / i - 8%, c / i - 61%, limf. - 22%, mon. - 7%.

Biochemical blood test: Society. Protein - 72.9 g / l, albumin - 39.3 g / l, globulins - 33.6 g / l, total. Bilirubin - 14.5 μmol / l, creatinine - 68.0 μmol / l, urea - 5.0 mmol / l, glucose - 4.22 mmol / l, AlT - 30.6 units / l, AcT - 28, 8 units / l, TG - 1.59 mmol / l, HDL - 0.61 mmol / l, LDL - 7.0 mmol / l, cholesterol - 5.4 mmol / l, IA - 7.8, alpha amylase - 52.0 U / ml, urinary to-that - 219.0 µmol / l.

Total urine analysis: quantity - 200 ml, color - c / f, beats. Weight - 1010, prozr., Protein - traces, sugar, acetone, bile. pig-you-otrits., p-tion - kitty, leykots. - 2-4-6 in p / sr., Eritr. - 20-30-50 p / sp., Epit. square - in the sky. que.

Radiography of OGK No. 1904-5 On the R-gram of OGP in 2 pr-y the pulmonary fields are transparent, the roots have a fuzzy structure, the sinuses are free. Sog - moderately expanded left ventricle. There are signs of osteopathy and deforming spondylosis of the cervical and thoracic spine. Zakl .: Khr. bronchitis. Consult a neurologist.

Ultrasound of the abdominal organs. Conclusion: Diffuse changes in the liver and pancreas.

Ultrasound of the kidneys. Conclusion: Solitary cyst of the right kidney 45x42 mm. Sinus cyst of the right kidney 26x24 mm. Solitary cyst of the left kidney 6-22 mm.

Ultrasound of the thyroid gland. Conclusion: Diffuse enlargement of the thyroid gland.

ECG No. 396 of 10/20/09. Conclusion: Sinus rhythm with a heart rate of 75 per 1 min. EOS to the left (angle alpha = + 10 °). Transition zone _four . Compensated atrial hypertrophy, left ventricular hypertrophy, indirect signs of incomplete blockade of the right leg of P. His. Signs of chronic subendocardial myocardial ischemia. РЦ = 0.19, Р = 0.12, ЦЯ8 = 0.10, ЦТ = 0.37 (N. = 0.30).

Consultation of the neurologist. Neurologically: Ό = 8. normal eye reflexes, photoreaction preserved, full movement, no nystagmus, instability and no vertigo, steady in Romberg’s posture, no pathological and meningeal pathological symptoms, moderate tactile and pain sensitivity, more pronounced in the extremities, decrease in muscle strength (up to 60%) in the limbs, in the muscle groups of the flexors and extensors, there is no marked movement disorder, the muscle tone is reduced. Paresis and paralysis is not. According to electromyography, there are signs of pronounced disorders in neuromuscular conduction in all muscle fiber groups.

Ophthalmologist: Visual acuity in both eyes 1.0. Fields of view without pathology. Signs of mild internal 31 018848 ч riokrapny hypertension. ECHO Encephaloscopy - displacement of the median structures was not detected.

Diagnosis: Arnold Chiari syndrome in the stage of subcompensation. Spinal cord hypoplasia. Cerebellar syndrome. Dysmetabolic polyneuropathy, mixed form. An additional blood test was carried out on the level of specific anti-tissue antibodies-autoantibodies to the tissues of the internal organs in a given patient according to the decision of the council of doctors. The patient's serum was studied, which remained after conducting biochemical studies (0.4 ml of serum was used). The protocol with the results of the study is given below in table. eight.

Table 8

The protocol for assessing the levels of detected antibodies, autoantibodies and the state of the tissues of the organs: 10.21.08; patient: D., 38 years old, diagnosis: Arnold Chiari syndrome (examination)

The condition of tissues of 7 organs was investigated. High levels of anti-tissue antibodies and antibodies to the thyroid gland, thymus (thymus), and liver were detected. The level of anti-tissue antibodies-autoantibodies to the kidney tissue indicates the beginning of an acute process, and the revealed titer value of 1: 1024 to the brain tissue indicates its severe damage, which began the process of lysis and rejection of the tissue. Revealed signs of chronic damage to the tissue of the thyroid and thymus glands, liver. Acute damage to the kidney tissue. Severe damage to the tissue of the white matter of the brain, accompanied by its partial lysis and rejection began. The revealed value of this indicator is not a favorable prognostic sign and may pose a threat to the life of the whole organism. 32 hours after the study, a sharp deterioration in the patient’s general condition began, requiring resuscitation and intensive care measures. The patient was transferred to the intensive care unit and despite ongoing activities and intensive therapy with increasing signs of renal-hepatic failure after 120 h there was a lethal outcome. This clinical observation confirms the possibility of obtaining laboratory data on the state of health and the level of tissue damage. Illustrates the possibility of detecting early signs of tissue damage and damage to organs in which other methods of laboratory and instrumental diagnostics do not reveal pronounced disorders in the internal structures, organs and systems of the body. The ability to detect early signs of emerging health disorders in internal organs in patients without clinical signs of disease using substances according to claim 1, obtained by the method according to claim 6, applicable to identify patients at risk of developing complications during mass vaccinations to prevent development of these complications.

Positive effect.

A positive effect in the diagnosis is achieved by solving a number of tasks:

the expansion of the range of substances determined in the laboratory that directly assess the state of health of body tissues;

increasing the sensitivity of the obtained antigens, the possibility of obtaining markers of the native type and their use in liquid-phase or solid-phase immunoassay methods, in manual, semi-automatic or automatic laboratory analytical systems.

expansion of the working range of concentrations, which allows you to do without diluting the sample and samples of biological fluids;

reduction of time, increase of efficiency, accuracy and predictive value of research results when conducting analysis using hardware;

the ability to create diagnostic kits on solid carriers that do not require special

- 32 018848 instruments and hardware, allowing to apply the method in non-laboratory or home conditions;

The implementation of the method for the simultaneous determination of several substances, in one biological sample taken from one individual, allows you to create an idea of the overall health of the whole organism, assessed by the degree of regeneration levels or tissue damage of the main organs or body systems.

The claimed technical solutions can be applied not only in mammals, but also in other objects of the animal world (reptiles, amphibians, birds, etc.), which allows them to be used in veterinary medicine of objects of the animal world - not mammals.

Substances with improved (other) properties do not have a direct lytic effect on the tissue and do not immediately cause a hyper-hypergene reaction, tissue lysis and death of the individual. The inventive substance, obtained by a technically different method described in the invention, has new properties, manifested in the fact that it can simultaneously exhibit the properties of the direct antigen of the tissue, promoting the production of antibodies that, due to the properties of the partial organ and species syngene to the original tissue, are anti-fabric ( antiorganic autoantibodies of the body, participating in acute and chronic reactions of the body, in which the immune (or autoimmune) system manifests its reactivity. Antibodies (autoantibodies) of the immune (autoimmune) response, produced in response to a substance, can cause local lysis (damage and utilization) of a part of an individual's organ tissue, without disturbing its vital functions and not leading to death of the whole organism. The same substance can and should be used as a laboratory control marker for assessing the effectiveness and depth of exposure to tissue. The substance has the ability to partial and isolated lysis (according to electron microscopy).

New substances do not act directly, but indirectly, so they can be used as drugs, including for the treatment of cancer. Substances are universal - they are diagnostics and drugs (one substance of a substance according to claim 1 is suitable for use as a pharmacological composition and for implementation as a diagnostic test system). Such properties of the substance allow to realize the dream of many doctors about the individualization of the possibilities of diagnosis, medication and therapeutic effects. In addition to this, the claimed biotechnology, subject to the relevant ethical norms and rules, allows us to consider the object of the upcoming diagnosis and treatment, as well as a possible source of the original ingredients (initial tissue) for the substance.

Thus, in the proposed technical solutions are given the main characteristics of the IIA, the components of the syngeneic composition. Identified, established, systematized and fixed in the lines of tissue compositions are signs of organ and species alignment of tissue obtained from a specific allogeneic tissue of an individual of the same species for a similar organ of all individuals of the same species. Antigenic similarity was established and differences in the lines of syngeneic compositions obtained from antigenically different tissues of internal organs in various animal and human species were determined. Criteria have been determined and a new symptom of specific immune activity has been identified in the substances that make up the syngeneic composition and that provide high specificity and result of the reaction. The method of using syngeneic compositions as direct markers of tissue damage levels of the investigated internal organs, laboratory methods for detecting quantitative and qualitative damage levels of the organs under study, and an assessment method with the ability to predict the state of health or illness of the organ (s) and the organism as a whole are given. The characteristic and classification of each line of syngeneic compositions obtained from the tissues of the internal organs of an animal or human is given. The use of lines of syngeneic compositions for the production of new types of laboratory diagnostics - direct immunoactive markers of tissues of internal organs is disclosed.

The use of markers obtained from the syngeneic composition of the tissue allows for express diagnostics and to identify the health conditions, injuries or diseases of the internal organs of an individual that arise in it without differentiating between various forms of diseases, both infectious and non-infectious. Declare IAV can be used as a basis for creating tools for high-precision and early diagnosis of cancer, as well as medicines and systems for the treatment of cancer.

Claims (14)

CLAIM 1. Biotechnological product, which is an immunologically active dehydrated protein substrate with disrupted internal hydrophilic bonds, obtained from the original tissue of an individual's organ, characterized by organ and species antigenic similarity to the protein of the original tissue, while deviating to it the individual antigen properties and autoantigen having a specific activity against antibodies and antibodies in biological fluids of other individuals about Nogo biological species, wherein the macromolecule comprises a protein product of the substrate in the range of molecular weight 10-90 - 33 018848 kDa, in which, during denaturing electrophoresis, specific areas are detected in the form of at least two major bands corresponding to the values of the molecular weight scale in the range 9.0-11.0 and 59-61 kDa. 2. Biotechnological product according to claim 1, characterized in that it contains at least one immunoactive syngeneic epitope of the original organ and species of an individual with antigenic autoantigenic properties. 3. Biotechnological product according to claim 1, characterized by the presence during high-performance liquid chromatography of two or three dominant fractions of alkaline components, weakly interacting with the anionic column at pH 7.2, and differing in charge of acid major components eluted in the middle and at the end the chromatography process, which appear on the chromatography curve as peaks corresponding to the ranges of 33.5 ± 2.5 and 52.5 ± 2.5 tAy for the two dominant fractions of alkaline components or the corresponding ranges Onam values of 17 ± 2,5; 33.5 ± 2.5; 52.5 ± 2.5 for the three dominant fractions of alkaline components. 4. The method of obtaining immunologically active biotechnological product, including sampling of the original tissue, its typing, repeated freezing at a temperature of (-24) - (- 28) ° C and thawing, while thawing is performed by exposure to microwave radiation at a frequency of 2450-2500 MHz removing the moisture formed during the thawing process to reduce the mass of the original tissue by 30-60%, the initial tissue obtained as a result of such processing, characterized by a syngality to the species and organ antigenic codes of the original tissue and devoid of individual antigens nnyh individual properties of the original tissue, minced, homogenized and extracted for decontaminated Grasse, resulting supernatant was subjected to standardization of 20-25 mg protein per 1 ml and freeze-dried. 5. The method according to claim 4, characterized in that tissue disinfection is carried out by adding 0.5% aqueous formalin solution to the tissue homogenate to 5-10% of the total starting material, followed by mixing and exposing the mixture for 4-6 hours. 6. The method according to claim 4, characterized in that the microwave effect is carried out at a radiation source power of 600-1000 W for a time that is set at the rate of 10-50 s per 100 g of the mass of the original tissue, while in the process of microwave exposure These parameters are set based on the condition of preventing the initial tissue from heating to a temperature of 41 ° C. 7. The method according to claim 4, characterized in that the microwave effect is carried out during the time at which the water crystallized on the surface of the frozen tissue becomes liquid. 8. The use of a biotechnological product according to claim 1 as a marker of tissue of the internal, transient or transplanted organs of the individual. 9. Biozond, comprising a solid phase coated with a biotechnological product on its surface according to claim 1. 10. Biobond according to claim 9, characterized in that the biotechnological product is applied to the solid phase as a series of dilutions in the concentration range from 12.5 to 200 μg of dry matter per 1 ml of saline. 11. Biobond according to claim 10, characterized in that a set of series of biotechnological products obtained from different tissues of an individual's organs is applied to the solid phase. 12. A method for diagnosing the condition of a tissue of an internal organ of an individual, including taking a sample of a sample of a biological fluid of an individual, ensuring contact of a biological fluid with a biotechnological product according to claim 1 or a bioprobe according to claim 9 in a series of dilutions of the sample of the test fluid within a titer of 1: 4 up to 1: 2048, the definition in the titers of the interactions between antibodies and / or autoantibodies found in the diluted sample of the biological fluid of the individual and the biotechnological product according to claim 1, with the conclusion that According to the limiting positive result of the interaction in a series of dilutions of the sample of the test liquid, if the interaction is found within the dilution of the test sample within the values of [1: 4-1: 8], they conclude that the tissue is healthy; within the values [1: 16-1: 32], conclude that there are insignificant tissue damage, within the values [1: 64-1: 128] they conclude that there is damage to the tissue typical for chronic inflammation, within the values [1: 256-1: 512] conclude that there is damage to the tissue that is typical for acute inflammation, within the range of [1: 1024-1: 2048] they conclude that there is damage to the tissue that is typical for lysis and / or rejection of the tissue. 13. A pharmaceutical composition comprising a solution comprising a biotechnological product according to claim 1 and an acceptable solvent, while for parenteral administration 1 ml of the solution contains 100 μg of the substance. 14. A method of treating diseases of the tissues of the organs of an individual, caused by the uncontrolled growth of cells of a tissue of a malignant or benign nature, comprising administering to an individual parenterally by uncommon the pharmacological composition of claim 13 containing a biotechnological product - 34 018848 according to claim 1, starting with a dose of 50 μg of the substance, followed by an increase to the amount that provides sensitization of the body and the response of the body’s immune system to the individual, as judged by the formation of anti-tissue antibodies-autoantibodies to the substance administered according to claim 1, with this introduction of the pharmacological composition is carried out by monitoring the level of formation of antibodies-autoantibodies in the biological fluids of the patient, the sampling of which is carried out after each administration of the composition, the number of injections limit the antitelautoantitel titers reaching levels lying in the range from 1: 256 to 1: 512.