Vaccine for prevention or treatment mediated t - cell pathology or unregulated replication of clones of t - cells, isolation of the vaccine, the method of diagnosing or predicting susceptibility to rheumatoid arthritis or multiple sclerosis, the method of prevention or treatment of rheumatoid arthritis or multiple sclerosis and containing the sequence sgdqggne peptide, which is an agent for the detection, prevention or treatment of multiple sclerosis

 

(57) Abstract:

The invention relates to medicine, in particular to immunology. The proposed vaccine for the prevention or treatment mediated T-cell pathology or unregulated replication of clones of T cells in the mammal contains an active ingredient and pharmaceutically tolerable environment as the active substances it contains the receptor of T cells or a fragment corresponding to the receptor of the T cells present on the surface of mediating the pathology of T cells, or antiidiotypic antibodies, which is the internal image of this receptor or its above-mentioned fragment, and an active substance taken in immunogene effective amount. It is prepared by obtaining clones of T-cells, causing mediated T-cell pathology, determine the amino acid sequence of the receptor T-cell clones of T cells associated with the pathology of the selection of data segments receptors of T cells, characteristic for these receptors T cells, but not for receptor T cells, not associated with this pathology, and selection of these sequences, those sequences which are capable of inducing immunogenic re is of being diagnosed with or predicting the susceptibility of an individual to rheumatoid arthritis, which is that it includes the detection of T cells containing variable plot-threads MSNQVLCCVVLCFLGANTVDGGITQSPKYLFRKEGQNVTLSCEQNLNHDAMYWYRQDPGQGLRLIYYSQIVNDFQKGDIAEGYSVSREKKESFPLTVTSAQKNPTAFYLCASS or its fragment in a sample of the individual, and abnormal expression of T-cell containing such a plot indicates rheumatoid arthritis or susceptibility to rheumatoid arthritis, as well as a method of prevention or treatment of rheumatoid arthritis, which is that it includes the prevention of the binding of the receptor of T cells containing a sequence MSNQVLCCVVLCFLGANTVDGGITQSPKYLFRKEGQNVTLSCEQNLNHDAMYWYRQDPGQGLRLIYYSQIVNDFQKGDIAEGYSVSREKKESFPLTVTSAQKNPTAFYLCASS, or receptor T cells, containing mainly the sequence SGDQGGNE, with his partner, and a method of preventing or treating rheumatoid arthritis in an individual, which is that it includes a cytotoxic or cytostatic treatment of T cells, MSNQVLCCVVLCFLGANTVDGGITQSPKYLFRKEGQNVTLSCEQNLNHDAMYWYRQDPGQGLRLIYYSQIVNDFQKGDIAEGYSVSREKKESFPLTVTSAQKNPTAFYLCASS, or T cells, containing mainly the sequence SGDQGGNE in the individual. 5 C. and 16 h.p. f-crystals, 10 PL.

The invention relates to the immune system, in particular to a vaccine for prevention or treatment mediated T-cell pathology or unregulated replication lonamin arthritis or multiple sclerosis, the method of prevention or treatment of rheumatoid arthritis or multiple sclerosis and containing the sequence SGDQGGNE the peptide, which is an agent for the detection, prevention or treatment of multiple sclerosis.

Higher organisms are distinguished by the immune system, protecting them from possible invasion of harmful substances or microorganisms. If the substance is classified as an antigen enters the body and if it ozyvaetsya as foreign, the immune system triggers and mediated antibody response and cell-mediated reaction. Cells of the immune system, denoted as B-lymphocytes or B-cells produce antibodies that specifically recognize foreign substance and communicating with them. Other lymphocytes, called T-lymphocytes or T-cells, and cause, and regulate the cell-mediated reaction, ultimately leading to the destruction of the antigen.

In cell-mediated reactions are involved in different types of T cells. Some of them induce specific clones of B-cell proliferation and production of antibodies specific for the corresponding antigen. Others recognize and destroy cells that represent foreign antigens on their surface. Certain T cells system is accurately controlled, however, deviations are not rare. In some cases, the immune system is not working properly and responds to the component of the host, as if he were a stranger. This reaction leads to autoimmune disease, which is reflected in the fact that the body's immune system attacks the tissue of the host. T cells as key regulators of the immune system, directly or indirectly cause such autoimmune pathology.

Assume that numerous diseases are the result of autoimmune mechanisms. The most famous of these diseases - rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis, diabetes type I, myasthenia gravis and common bladderwort. Millions of people worldwide suffer from autoimmune diseases, and associated with these diseases costs, including the treatment itself, incidental expenses and loss of labor productivity are billions of dollars each year. There is currently no means for effective treatment of such autoimmune pathologies. You can usually treat only the symptoms, and the disease progresses, often leading to a serious weakening of the patient or his death.

In other cases, the lymphocytes multiply excessively and uncontrollably education is Auda T-cells, these tumors are referred to as lymphoma T-cells. Like other malignant tumors and lymphoma T cells effectively treat hard.

Thus, already for a long time need effective tool for the treatment or reduction of the intensity of pathologies mediated by T-cells. In the ideal case, such a treatment would rather control inappropriate response of T cells than only reducing the intensity of symptoms. The present invention satisfies this requirement and at the same time has side benefits.

The first object of the present invention is a vaccine for prevention or treatment mediated T-cell pathology or unregulated replication of clones of T-cells in mammals, containing pharmaceutically tolerable environment and the active substance representing the receptor of T cells or a fragment corresponding to the receptor mediating the pathology of T-cells present on their surface, or antiidiotypic antibodies, which is the internal image of this receptor T cells or the above-mentioned fragment, and an active substance taken in immunogene effective amount.

Under the concept of "mediated T-cell is onetom pathology. This notion here covers and disease, directly mediated by T-cells and disease, e.g. myasthenia gravis, which primarily occur in the violation caused by the binding of the antibody, as well as diseases caused by an inappropriate response of T cells leading to the production of such antibodies. I.e., this notion covers and autoimmune disease mediated by T-cells, and unregulated replication of clones of T cells.

The following turnover "in the main sequence" in connection with the sequence of amino acids should be understood or described sequence, or other sequence in which added, omitted or replaced by separate units, and these changes do not significantly affect the ability of the sequence to induce an immune response against the desired sequence receptor T cells. Thus, the area described by immunizing sequence can be used always when he is sufficiently characteristic for the desired receptor T cells in order to induce an effective immune response against the desired receptors of T cells, but not against unwanted receptors of T cells. Such variations posively you can check e.g., by immunization of mammals.

The term "fragment" in the following covers fragments in combination with additional sequences or parts, in which, e.g., the peptide is linked to other amino acid sequences or with a carrier, or fragments associated with them. The terms "fragment" and "peptide" may therefore be used in this text as interchangeable concepts, because the peptide will be the most common fragment of the receptor of T cells. Each fragment according to the invention may have a modified sequence as described above in section "main sequence".

When in the following we are talking about the "fragment or segment of the receptor T cells", then this does not mean that they must be derived intact receptor T cells. Such fragments or sections can be performed using various known methods, e.g., by manual or automated peptide synthesis or by cloning.

In the following in connection with the relationship between the proposed fragment peptide and sequences of receptors of T cells, the word "corresponding" means that the peptide fragment has the sequence aminoquinolones reaction in the individual. However, the sequence is not necessarily identical with the sequence of the receptor T cells, as seen in examples II and III.

Under the words "immunogene effective" here refers to the number of receptor T cells or its fragment, which effectively causes an immune response for the prevention or treatment mediated T-cell pathology or unregulated replication of clones of T cells in the individual. Needless to say that this number can vary depending on the substance and of the individual, and many other factors. For example, in General a person for an effective immune response requires a large dose than the mouse.

As the availability of vaccine in a liquid state, the concentration of the active substance is about 1 μg - 100 mg / ml In the case of the proposed use of the vaccine in the solid state active substance take appropriate concentration.

In the following under "V17" refers to a specific variable plot-chain receptor T cells. V17 has the following amino acid sequence: MSNQVLCCVVLCFLGANTVDGGITQSPKYLFRKEGQNVTLSCEQNLNHD AMYWYRQDPGQGLRLIYYSQIVNDFQKGDIAEGYSVSREKKESFPLTVTSAQK NPTAFYLCASS. Hypervariable and binding sites are the most useful for accused particularly useful. Modifications of this sequence did not affect the ability of the receptor to act as an immunogen, inducing the desired immune response, are also covered by this term. Variable plot can be sawsan with any D - or J-segment receptor T cells. In addition, immunogene active fragments V17 also covered by the definition of "V17".

In the following under "partner (to link)" refers to a composition capable of interacting with the receptor of T cells. In General this structure is the main antigen tissue compatibility (MHC), but it can be any composition, with the usual binding receptor of T cells leading to activation or proliferation of T cells.

The term "ligand" in the following refers to any molecule that reacts with another molecule to form a complex.

The word "selectively binds to" the following should be understood to mean that the molecule binds to one type of molecules, but mostly not with others. In relation V17 "selectively binds to" include binding with receptors on T cells containing V17 , but mostly not with receptors on T cells that do not contain V17.

The immune system is g is DNAME agents can be, e.g., pathogenic factors, e.g., bacteria or viruses, and also modified its own cells, including infected with the virus cells, tumor cells and other abnormal cells of the host. All these objectives of the immune system is generally called antigens. In the recognition of antigen by the immune system immediately activates the immune mechanism for the destruction of the antigen, thereby protecting the health of the host.

The main types specific for a particular antigen immune response is a humoral (mediated by antibodies) and cellular (cell-mediated) immunity. Each of these immunologic mechanisms embarks activation of helper (CD4+) T-cells. Data CD4+ T-cells stimulate B-cells capable of antibody synthesis by binding to antigens, with the result that they proliferate and secrete antibodies. Secreted antibodies bind to antigens and facilitate their destruction by other immune mechanisms. Similarly CD4+ T-cells give stimulating signals cytotoxic (CD8+) T-cells that recognize and destroy the corresponding cells (e.g., infected with the virus host cell). Thus, activation of CD4+ T cells is Cescau for a particular antigen activated CD4+ T cells, it is important in any attempt selective modification of immunological actions.

The specificity of T cells against certain antigens is provided by receptor T cells, expressed on the cell surface. Receptor T cells is a heterodimeric glycoprotein consisting of two polypeptide strands, each of which has a molecular weight of approximately 45 base pairs (p. O.). There are two forms of the receptors of T cells. One consists of alpha-strands and beta-strands, and the other from a gamma-thread and the Delta thread. Each of these polypeptide strands receptor T cells encoded with a specific genetic locus containing complex discontinuous segments of genes, representing, e.g., variable (C) linking (J) or not change (C) plots. Beta - and Delta-thread contain an additional element, denoted as diversicornis (D) segment. (Because diversicornis segments (D) and items are found only in certain genetic loci receptors of T cells and the polypeptides, below they are given in parentheses in order to show that these sites are only in their respective threads receptors of T cells. Thus, V(D)J refers or to the sequence of the VDJ filaments with a D-phase, the segments will regroup and form a functional gene, determining the amino acid sequence of the receptor T cells expressed the cell. Because the stock V-, (D) and J genes, which can be converted contains many members and due to the fact that the individual elements of this reserve can regroup with any combination of common stock receptors of T cells is highly diversicornis and capable of linking countless partners that can occur on the body. However, each T-cell has only one molecule of the T-receptor, which mostly, if not completely, determines the specificity of this T-cell relative to its partner.

Model animals has contributed significantly to the understanding of the immunological mechanisms of autoimmune diseases. One of the models is experimental allergic encephalomyelitis (EAE), an autoimmune disease of the Central nervous system that can be called in mice and rats by immunization with myelin basic protein (ICBMs). The characteristics mentioned diseases the following: clinical paralysis and mild malnutrition, and histologically - perivascular mononuclear infiltration of the parenchyma cells of the Central nervous system. Pathogenesi specific in respect of the basic protein of myelin T cells were isolated from animals, suffering from experimental allergic encephalomyelitis, and amplified by successive cultivation. After stimulation in vitro myelin basic protein data T cells immediately called experimental allergic encephalomyelitis, when their adaptive transferred into a healthy host. It is important that the data-inducing experimental allergic encephalomyelitis with T-cells specific not only in terms of this particular antigen (a basic protein of myelin), but usually against a single epitope on the antigen. All of this means that small amounts of autoaggressive T cells are responsible for the pathogenesis of experimental allergic encephalomyelitis.

The analysis of the receptors of T cells, inducing experimental allergic encephalomyelitis, revealed that the data structure associated with the disease receptors is reduced heterogeneity. During the analysis, which explored 33 interacting with myelin basic protein T-cells, found only two contain variable parts of the segment alpha-strands and a single segment alpha-strands containing the binding site. In this population of T cells t the and contain variable parts of the segment of the beta-strands and two segments, contains binding sites. More importantly, about 80% of clones of T cells at the V-D-J-parts of beta-strands had identical amino acid sequence. These results confirm that the receptors of T cells with a similar specificity antigens also have the same structure, and show that the receptor of T cells is a suitable target for immunotherapeutic strategies in the fight against pathogenesis of experimental allergic encephalomyelitis.

Various methods tried to use the specificity of autoaggressive T cells relative to the antigens to develop strategies for the treatment of experimental allergic encephalomyelitis. For example, used passive cottage monoclonal antibodies specific for receptors on the surface of inducing experimental allergic encephalomyelitis with T-cells. In the case of a murine model of experimental allergic encephalomyelitis in the infusion of a monoclonal antibody specific for V,8, representing more variable plot beta-strands, used in relatively specific basic protein of myelin T-cells, the susceptibility of mice to the subsequent individual, and urban and other, Cell, N 54, pages 577 - 592, 1988). This protection was achieved in the case of experimental allergic encephalomyelitis in rats by using monoclonal antibodies capable of interacting with unidentified idiotypical determinant of receptor T-cell-specific relative to the basic protein of myelin T-cells (see Barnes and others, J. Exp. Med., N 169, pp. 27 - 39, 1989 ). Although therapy passive antibodies apparently has a positive effect on susceptibility to experimental allergic encephalomyelitis, it is associated with potential problems. Achieved protection disappears, and therefore requires re giving antibodies. When multiple infusions of antibodies, however, increases the likelihood that the host develops an immune response against fed antibodies, in particular then, if they got in a xenogeneic animal. In addition, the response of antibodies to pathogenic clones of T-cells represents only one part of the entire immune response and prinebregaete possible role of cellular immunity in overcoming autoreactivity.

The role of cellular immunity in reducing the activity of autoaggressive T cells in experimental allergic encephalomyelitis regulatory T cells, then again given to the owner under immunotherapy. E.g., most recently allocated clone of CD8+ T cells from convalescent rats, in which by adoptive transfer of specific regarding the basic protein of myelin line CD4+ T cells caused experimental allergic encephalomyelitis (see San and others, Nature, N 332, pages 843 - 845, 1988). This clone CD8+ T cells in vitro showed cytolytic activity against containing CD4+ T cells used for the induction of disease. In addition, adoptive transfer of this CTL clone reduced the susceptibility of the respective rats on the subsequent submission of the basic protein of myelin. The leader and others (see Science, No. 239, page 181 -183, 1988) also highlighted the clones of CD8+ T cells with suppressive activity relative to the cause of experimental allergic encephalomyelitis with T-cells. This clone CD8+ T cells were isolated from rats vaccinated with attenuated clones of T-cells that cause the disease, and, although he did not show zitoliticescoy activity in vitro, it was capable of suppression is dependent on the basic protein of myelin proliferation of T cells, inducing experimental allergic encephalomyelitis. Although these analyses show that the containing CD8+ T cells capable of CD8+ CTL will play an important role for long-term resistance recovered rats because Sedgwick and other (Eur. J. Immunol., N 18, PP 495 - 502) clearly showed that the deletion containing CD8+ cells with monoclonal antibodies does not affect the progress of the disease or recovery.

In the above experiments, Sana and others, and the Leader and other cottage regulatory T cells leads to the overcoming of the important problems of passive antibody therapy, because it allows you to achieve long-term regulatory response in vivo. However, it requires cultivation in vitro using attenuated T-cells that cause the disease in order to develop clones of such regulatory T cells, which is a costly and time-consuming process. In addition, in the case of a person because of the identical, the main antigen tissue compatibility, this requires a high degree individualizovane therapeutic strategy, because for each individual patient must obtain regulatory clones, which are then only give it to the patient in order to exclude a possible graft-versus-host.

Immediate vaccination attenuated clones of T-cells that cause the disease, used for therapy of experimental allergic encephalomyelitis. T-cells, relatively specific about the practical fixing and then they were used for vaccination of uninfected rats. In some cases vaccinated animals developed resistance with respect to subsequent attempts to elicit experimental allergic encephalomyelitis (Leader and others, see above, as well as in Cohen and Weiner, Immunol. Today, N 9, pp. 332 - 335, 1988). However, the efficacy of this vaccination is variable, and the degree of protection varies considerably. T cells contain many antigens that elicit an immune response, when the whole T-cell is used as a vaccine. It showed Offner and others (J. NeuroimmunoL, No. 21, pages 13 - 22, 1989), who showed that immunization with the use of a T cells contributes to allergic reactions of the delayed type, defined on the ear swelling, data on T-cells, which increases with the number of vaccinations. However, a clear allergic reactions of the delayed type installed in protected and unprotected animals. Rats showed a similar response after vaccination and causing encephalitis T cells and control T cells. Conversely, specific vaccination relative to purified protein derivative of the T-cells of relatively specific purified protein derivative line caused an allergic reaction of the delayed type and cells of the vaccine, and nazyvausia disease cells and control cells, defined on the allergic reactions of the delayed type (which is a measure of cell-mediated immunity), indicates that many data antigens T cells trigger an immune response. Thus, the disadvantages of vaccination attenuated T-cells that cause the disease are the lack of specificity regarding protective antigen on the surface of the T-cells, as well as inconsistent results regarding immunity against the antigen. With regard to the application of this method to treat people, it is as laborious as described above, the infusion of CD8+ cells, and as this last method requires individualizovani therapy.

The present invention provides an effective way immunotherapy of pathologies mediated by T-cells, including autoimmune disease, not associated with those problems that are associated with the above known methods of treatment. By vaccination and not by passive villas heterologous antibodies mobilized own immune system of the host to the suppression of autoaggressive T cells. Thus, suppression is permanent and can cover any and all immunological mechanics is the suppression, achieved by passive use of monoclonal antibodies or clones of regulatory T cells.

Because the proposed vaccine used to combat autoimmune diseases, they contain receptors of T cells mediating autoimmune disease. Vaccines can be either integer receptors of T cells, mainly purified from the clones of T-cells, or individual threads receptors of T cells (e.g., alpha -, beta-, gamma-threads), or parts of such filaments, or separately, or in combination with each other. The vaccine can be either homogeneous, e.g., to include only one peptide, or contain more than one type of peptide, each peptide refers to different parts of the receptor. In addition, these peptides can occur from different receptors, when both receptors contribute mediated T-cell pathology.

According to a special form of execution of the invention immunizing peptide can have an amino acid sequence SGDQGGNE, if it is intended to serve for the treatment of multiple sclerosis. Any immunogenic portion of this peptide can be effective. Thus, it is possible to replace amino acids without loss of immunogenicity of the peptide. In addition, this peptide can be associated with the carrier, however, the purpose of man, suffering from rheumatoid arthritis, you can use the receptors of T cells or fragments of receptors containing V17 , for the treatment or prevention of this disease. Caused by the human immune response can neutralize or destroy T-cells with V17, and thus to prevent or to treat their harmful effects. In addition, to the extent V17 there are also receptors for pathogenic T-cells, mediating autoimmune disease in General, the proposed vaccine can be used to deal with other autoimmune diseases.

The word "mostly clean" means that the receptor T cells are essentially free from other biochemical components with which it is normally associated in nature. Alternative vaccines contain peptides of different lengths, corresponding to the receptor of T cells or parts thereof. The peptides can be obtained either by synthesis or by recombination using known for specialist methods. Preferably a peptide vaccine corresponding sections of the receptors of T cells, which this receptor differs from other, non-pathogenic receptors. These specific areas can be separate(ohms) sections(s) of the respective polypeptide strands Renou response to T-cells, contains the determinant.

Vaccines give in if the owner shows an autoimmune reaction, or is in such danger. Accurate clinical diagnosis of certain autoimmune diseases justifies giving the necessary specific for a given disease vaccines containing receptors of T cells. Preventive cottage justified where autoimmune mechanisms precede the beginning of the open clinical disease (e.g., diabetes type I). Thus, these vaccines can be before disease prophylaxis be used in people with a family history of a corresponding disease, and who, judging by reliable performance, are at risk of the disease.

Containing receptors T-cell vaccines can be used with many possible formulations with pharmaceutically tolerable environment. In case of a short peptide of the latter may be associated with a carrier, e.g., isolated from the platter by hemocyanin, in order to increase its immunogenicity. The vaccine can be given in combination with adjuvant (there are several known adjuvants). After the initial immunization the vaccine can be submitted amplifier. Vaccine appliciruut known methods in doses that are sufficient for the purpose peptides can find in the following way. Clones causing disease T-cells that interact with antigens that want to fight, isolated from infected individuals. Such T-cells, preferably in a place where there was a strong autoaggressive activity, i.e., in place of the pathological changes in the case of ordinary water from the Central nervous system in the case of multiple sclerosis, synovial fluid or tissue in the case of rheumatoid arthritis or from the appropriate blood of an infected individual. Then determine the sequence of amino acid receptor gene autoaggressive T cells. Then vaccino used in this way, you can choose polypeptides corresponding receptors of T cells or portions of receptors available, in particular in causing disease T-cells (compared with non-pathogenic T-cells).

Instead, the vaccine may contain antiidiotypic antibodies, which is the internal images of the above peptides. Methods of preparation, selection and application of such antiidiotypic vaccines are known (see, e.g., Eichmann and others, CRC Critical Reviews in Immunology, N 7, PP 193 - 227, 1987).

Mediated by T-cells with malignant pathology etiolate, and caused T-cell lymphoma is a pathology amenable to this treatment. The application of this technology in the treatment of T-cell lymphoma can be done in essentially identical to the method. However, this technology is used rather in T-cell proliferative disease, because it is easier to make the selection of pathogenic T cells. After selection of clones of technology carried out the following way. In particular, determine the sequence of amino acid receptor gene in T-cells T-limfoma and determine the appropriate sections of these receptors, which are used as vaccines. Vaccines can contain either a simple or complex peptides, and can be applied in a known manner into pharmacologically tolerable composition with or without adjuvant.

Multiple sclerosis

Causing multiple sclerosis T-cells is still not identified, although because of the clinical and histological similarities of multiple sclerosis and experimental allergic encephalomyelitis was suggested that T-cells that interact with myelin basic protein, play a role. In experimental allergic encephalomyelitis in rats and mice encephalocele T cells, wsimages is as VDJ-plot-threads, despite known differences in restriction of the main antigen tissue compatibility and antigenic specificity of the basic protein of myelin. The present invention is based on the discovery that the line T-cells that interact with myelin basic protein and derived from patients with multiple sclerosis, has-thread-receptor T cells with VDJ sequence of amino acids corresponding to the sequences on the threads of the T-cells that interact with myelin basic protein and cause pathogenesis in experimental allergic encephalomyelitis, which is a model of multiple sclerosis in animals. This line is specific for a different epitope of the basic protein of myelin. This discovery shows that interacting with myelin basic protein T cells participate in the pathogenesis of multiple sclerosis and that the receptor peptides similar to those described for the prevention of experimental allergic encephalomyelitis, can be used for the treatment of multiple sclerosis.

Rheumatoid arthritis

Rheumatoid arthritis is an autoimmune disease mediated by T-cells. The invention describes oligoclonal infiltrate the cells in the diseased tissue of all examined patients, their oligoclonality and cytotoxic activity of one of these T cells to synovial attached cells show a Central role containing V17 T-cells in the pathogenesis of rheumatoid arthritis.

The activated population of T cells in synovial tissue of patients with rheumatoid arthritis investigated by analyzing mRNA receptor T cells isolated from synovial T cells, positive relative to the receptor of IL-2 (IL-2R+). mRNA receptor T cells amplified in a known manner by chain reaction using Taq polymerase (PCR). This analysis revealed that oligoclonal V17 rearrangement enriched in the population IL2-R+, which indicates the probability of participation containing V17 T cells in the pathogenesis of rheumatoid arthritis. From the sample of synovial tissue were isolated clone of T cells containing CD4+, V17 and the fact that he has cytotoxicity in vitro relative to the synovial attached cells, confirms a direct part containing V17 T cells in rheumatoid arthritis.

As already indicated, the present invention provides an extremely important discovery, namely, that specific variable plot-threads receptor T cells, referred to as V17, closely tie the rheumatoid arthritis with the use described in this invention. The specialist will be able to apply in relation to rheumatoid arthritis therapeutic methods such as those described above in connection with experimental allergic encephalomyelitis methods.

Thus, a second object of the invention is a method of diagnosing or predicting susceptibility to rheumatoid arthritis or multiple sclerosis individual. In the case of rheumatoid arthritis, the method includes detection of T cells containing denoted as variable V17 plot-strands in the sample of the individual, and the presence of abnormal quantities containing V17 T-cells indicates rheumatoid arthritis or susceptibility to rheumatoid arthritis. Containing V17 T-cell can qualitatively or quantitatively be compared to a normal cell of the individual. This diagnosis can be performed by detecting part V17, which is not available in variable plot-threads receptors of T cells that are not associated with rheumatoid arthritis. V17 can be detected, e.g., by bringing V17 in contact with the detector ligand capable of specific binding with V17. Numerous detector ligands, e.g., associated with the enzyme antibody. Instead, for detection saturnotaku nucleic acids (see example IX).

In the case of diagnosing or predicting susceptibility to multiple sclerosis proposed method comprises detecting in a sample of individual T cells, which mainly sequence SGDQGGNE, and the presence of this sequence indicates multiple sclerosis or susceptibility to multiple sclerosis. The sequence can be detected, e.g., by bringing into contact with the detector ligand. Numerous detector ligands, e.g. associated with the enzyme antibody. Instead, for the detection of T-cells can also be used nucleotide probes complementary to the coding sequence of the nucleic acid (see example IX).

The third object of the invention is a method of preventing or treating rheumatoid arthritis or multiple sclerosis. In the case of rheumatoid arthritis, the method includes preventing binding containing V17 T cells with her partner. According to one form of the invention the binding is prevented by binding of the ligand with V17. According to another form of execution of the invention the binding is prevented by binding of a ligand with a partner V17. Linking can be prevented world is ivania.

According to another possible implementation of the invention the prevention or treatment of rheumatoid arthritis can also be done by cytotoxic or cytostatic treatment containing V17 T cells in the individual. According to one variant containing V17 T cells treated with a cytotoxic or cytostatic agent that selectively binds V17. The agent may be a radioactive or chemotherapeutic agent. Such linking and suitable for this purpose, the agents are widely known (see, e.g., Harlow E. and lane, Antibodies, a Laboratory Manual, Cold spring Harbor, Laboratory, 1988).

In the case of prevention or treatment of multiple sclerosis, the method includes preventing binding containing mainly the sequence SGDQGGNE T cells with her partner. According to one form of the invention the binding is prevented by binding of the ligand sequence. According to another form of execution of the invention the binding is prevented by binding of a ligand with a partner. Linking can be prevented with known methods, e.g. by linking the antibody sequence to the physical blocking of the binding.

According to another possibility th processing containing mainly the sequence SGDQGGNE T cells in the individual. According to one variant of the T-cells treated with a cytotoxic or cytostatic agent that selectively binds the sequence. The agent may be a radioactive or chemotherapeutic agent.

The fourth object of the present invention is a method of allocating vaccines for the treatment mediated T-cell pathology, including the production of clones of T-cells that cause this condition, the determination of the amino acid sequence of the receptor T-cell clones of T cells associated with the condition, the selection of data segments receptors of T cells, characteristic for these receptors T cells, but not for receptor T cells, not associated with this condition, and the selection of these sequences, those sequences which are capable of inducing immunogenic response in respect of these receptor T cells, thus taking the vaccine.

The fifth object of the present invention is containing a sequence SGDQGGNE peptide, which is an agent for the detection, prevention or treatment of multiple sclerosis.

The sixth object of the invention is the method according to PP. 25-27 claims.

Invented the myelitis rats

Female rats varieties Lewis (Charles river laboratories, Raleigh-Durham, USA) were immunized by filing in the paw of each of the rear legs 50 ál of the basic protein of myelin Guinea pigs in the form of an emulsion in full stimulator's adjuvant. The first symptoms of the disease usually found for 9-11 days after immunization. The degree of disease was assessed by using a scale with three sections: 1 - lame tail, 2 - weakness in the hind legs, 3 - paralysis of the hind legs. After the duration of illness by approximately 4 - 6 days most of the rats spontaneously recovered and was resistant to the subsequent induction of experimental allergic encephalomyelitis.

Example II

The allocation of vaccines

Vaccination was performed with peptide receptor T cells, the sequence of which is deduced from the DNA sequence of a beta-receptor gene in T-cells, prevailing among inducing experimental allergic encephalomyelitis with T-cells of mice varieties B10.PL/L. the DNA Sequence corresponds to the instructions Urbana and others (see above). The peptide containing nine amino acids and having a sequence of the VDJ-threads receptor T cells from mouse, synthesized by a known method. The sequence of this peptide is as follows: SGDAGGGYE. For all who eat a column chromatography on a column on a Sephadex G-25 (Pharmacia Fayne, Chemicals, Piscataway, new Jersey, USA) using as eluent 0.1 M acetic acid, and the solvent was then removed by lyophilization in two cycles. Part of the peptide using glutaraldehyde conjugatively with allocated from the platter by hemocyanine (LH) in the ratio of 7.5 mg of peptide per mg of hemocyanin. The conjugate is dialyzed using a saline phosphate buffer (RVS).

Example III

Vaccination against experimental allergic encephalomyelitis

Used in these experiments vaccines consist of free peptide with VDJ-section and of the peptide with VDJ-plot conjugated with allocated from the platter by hemocyanine. They were diluted in saline phosphate buffer and emulsiable equal parts, or (1) incomplete stimulator's adjuvant or (2) full stimulator's adjuvant, obtained by suspension 10 mg/ml killed by heat treatment of the dried Mycobacterium tuberculosis H37ra (Difco the laboratories, Detroit, USA) in incomplete stimulator's adjuvant. Emulsion gave a rats-females varieties Lewis age 8 - 12 weeks in total amount of 100 µl per animal (50 μl in each rear paw feet). Each rat was given 5 μg unconjugated VDJ peptide. The conjugate is selected from a saucer of hemocyanin and VDJ gave vdeu rat gave 50 mg of the basic protein of myelin Guinea pigs in full stimulator's adjuvant in the front paws. Animals were monitored every day, starting from the 9th day to discover the clinical symptoms of experimental allergic encephalomyelitis, and they were evaluated as described above. The results of the experiment are shown in table I (see the end of the description). They show not only that the vaccinated animal disease revealed less, but that in the diseased animals, the disease progressed less difficult and/or its start has been delayed. The degree of protection were different, as well as the composition of the vaccines, and vaccines containing the full stimulator's adjuvant, showed the greatest degree of protection.

Example IV

Vaccination against experimental allergic encephalomyelitis with VDJ peptides obtained from rats varieties Lewis

VDJ peptide used in the previous examples, was synthesized according to the sequence of molecules in the chain of the receptor of T cells found in the calling of experimental allergic encephalomyelitis with T-cells of mice varieties B10.PL. Data VDJ sequences are homologous, but not identical to the sequences found in T cells of mice varieties B10. PL. Peptides rats synthesized according to the DNA sequences, called Bernson etc. and Cliveden below, together with the sequence of mouse types B10.PL used in examples I and III (VDJ).

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Receiving, giving and evaluation of these vaccines was carried out as described in examples I - III follows, with the following exceptions. In vaccine compositions containing the full stimulator's adjuvant, and used 50 mcg separate VDJ peptide; never used vaccines in incomplete stimulator's adjuvant or vaccine containing conjugate selected from the platter by hemocyanine (LH). The control animals prior to submitting the basic protein of myelin (ICBMs) were not or were vaccinated emulsion phosphate salt solution (PBS) and complete the stimulator's adjuvant (CFA) in order to evaluate the protective effect of one adjuvant. The results are shown in table. II (see below).

In table II the results show that in unvaccinated control animals disease already in the 10th day. The disease manifested itself in severe paralysis and impairment continued for 4 to 6 days and then spontaneously stopped. Rats vaccinated with PBS-CFA, the disease proceeded in essentially the same manner as in the unvaccinated control animals. In contrast to some of the animals vaccinated with IR1, IR2 is abolitionist disease. However, in General vaccination VDJ peptides rats (IR1-3) was slightly less effective than vaccination VDJ peptide mouse (see example III). However, vaccination IR9b resulted in complete protection in all four animals in which it was investigated. Important is the fact that none of the four animals vaccinated IR9b, did not show any histological changes characteristic of the disease, which indicates that they were also prevented subclinical symptoms of the disease.

Example V

Vaccination with peptides specific regarding the variable segment

Specific for a family of V8 peptide genes investigated as a vaccine against experimental allergic encephalomyelitis. V8 is the most common family-thread, and it is used in encephalitogenic T-cells of rats and mice. The peptide was synthesized according to a unique sequence of DNA that is found in the gene V8 , and not in other genes of the rat, the sequence of which is shown Morris and others (Immunogenetics, No. 27, pages 174 - 179, 1988). The sequence of this V8 peptide, referred to as the IR7, the following:

IR7 DMGHGLRLIHYSYDVNSTE.

The effectiveness of this peptide was studied in eksperimentalnoi with full stimulator's adjuvant (CFA). Vaccination with incomplete stimulator's adjuvant (IFA) or a conjugate of peptide with allocated from the platter by hemocyanine (IFA) was not implemented. The results of these experiments are shown in table. III (see the end of the description).

The result of the vaccinations carried out by the peptide of the rat, similar to the results of vaccinations IR1, IR2 and IRS peptides in rats and mice. One animal was found reduced intensity and duration of the disease, and one animal was protected completely.

Example VI

Vaccination containing J-areas peptides

The peptide was synthesized according to the segment J-strands, TO found among receptors encephalitogenic T cells and in rats and mice. The sequence of this peptide, referred to as visa ir5, the following:

VISA IR5 RFGAGTRLTVX.

Efficiency JTA39 peptide was studied in experimental allergic encephalomyelitis in rats varieties Lewis (example I) described in examples II and III. 50 μg of the peptide was studied in full stimulator's adjuvant (CFA). Vaccination with incomplete stimulator's adjuvant (IFA) or a conjugate of peptide with allocated from the platter by hemocyanine (IFA) was not implemented. The results of these experiments are shown in table. IV (see below).

The results of the peptide. Two of the three animals were fully protected, and in the third animal disease onset was significantly delayed. In addition, in this animal the intensity of the disease was also reduced, although the disease had normal duration of 5 days. Important is the fact that both are fully protected animals did not show any histological symptom infiltration of T cells into the Central nervous system. This result shows that vaccination JTA39 very effectively calls the regulatory response aimed at encephalitogenic T cells. Even went undetected subclinical symptoms of the disease.

Example VII

Vaccination mixtures of peptides receptors of T cells

Vaccination was carried out with a mixture of peptides receptor T cells. This mixture contained 50 µg of peptides IR1, IR2 and IR3 (VDJ peptides three rats and peptide J39 rats).

The effectiveness of this mixture of peptides was studied in experimental allergic encephalomyelitis in rats varieties Lewis (example I) described in examples II and III. The peptides investigated in full stimulator's adjuvant (CFA). Vaccination with incomplete stimulator's adjuvant (IFA) or a conjugate of peptide with allocated from the blue is="ptx2">

The results of vaccinations performed by using a J39 peptide rats and three VDJ peptides, are almost as effective as the results of vaccination IR9b given in table II. All three animals were fully protected. Along with the absence of clinical symptoms of experimental allergic encephalomyelitis two of the three animals were completely free from histological symptoms of infiltration of T cells into the Central nervous system, and the third saw only two small hearth lymphocytic infiltration at the base of the spinal cord.

Example VIII

Vaccine against multiple sclerosis

Interacting with myelin basic protein T-cell human

Line interactive with myelin basic protein T-cells obtained from peripheral mononuclear blood cells (RVMS) of nine patients with chronic progressive multiple sclerosis and two healthy control people. Cells for 3 days were kept in culture with regular stimulation cleaned the basic protein of myelin person and irradiated autologous peripheral mononuclear blood cells, and then for 4 days in containing IL-2 environment.

Ampli is them with myelin basic protein T-cell

T cells were isolated from cultures obtained after logarithmic phase of growth, and received RNA, amplified V16-dimensional example and example C for the implementation of 55 cycles according to example IX.

Sequence-threads receptor of T-cells that interact with myelin basic protein

Using the example Cseq determined the sequence of the amplified V16-Merom genes threads receptor lines of T-cells that interact with myelin basic protein. The amplification product was purified using the gel was denaturiruet Foundation and by example Cseq determined its sequence. Of the five lines have been able to read the DNA sequence, which showed that the dominant clones of T cells obtained through long-term migration in vitro. One of the data sequences from the cell line Re (see tab. VI at the end of the description), had VDJ sequence of amino acids is the thread in which five of the first six and six of the nine in total identical residues with the corresponding residues VDJ sequence of amino acids of the thread stored in encephalogram T-cells that interact with myelin basic protein in experimental allergic ence is ruppirovka receptor T cells, found in the remaining four lines of T-cells that interact with myelin basic protein.

To determine the possible presence of similar sequences in the thread lines of the T-cells that interact with myelin basic protein, other multiple sclerosis patients was carried out by PCR amplification with degenerate (n = 1024) 21-nucleotide sample (VRe), corresponding to the seven amino acids of the sequence (see table VI). Was carried out by reverse transcription of RNA, which V16-dimensional example and example Cext amplified reaction in 20 cycles (stage I). 1 ál aliquot of sample product of these reactions stage I was reamplification 35 cycles examples VRe and Cint. 1 ál aliquot of sample product of these reactions was investigated by hybridization to Southern labelled32P-sample person. This study showed amplificatory product with 300 p. O. in cell lines of Re and in one other patient with multiple sclerosis, but not in T-cells that interact with myelin basic protein, control of individuals or lines and clones do not interact with myelin basic protein T-cells. The presence of this sequence in two of the nine investigated the STN, remains among encephalogram T cells in experimental allergic encephalomyelitis, its detection in T-cells that interact with myelin basic protein, patients with multiple sclerosis shows that the determinant of T-cells play a role in the pathogenesis of multiple sclerosis.

Immunogenic peptides having the sequence SGDQGGNE, can be obtained by the synthesis described in example II follows, and can be used for immunization of people described in example III method. Such immunization may lead to an effective immune response.

Example IX

The allocation of oligoclonal infiltrates II activated T-cells containing V17, Zinovii patients with rheumatoid arthritis

Obtaining T cells from the synovial tissue

Samples of synovial tissue obtained from patients in whom rheumatoid arthritis was confirmed by radiography, and which was treated with prosthetic joints. Using magnetic pearls and antibodies that interact with IL2-R ( IL2-R) person, in the following way got activated T-cells. Synovial tissue was digested at a temperature of 37oC for 4 h in RPMI medium with 10% serum fetal cow (FBS), containing the Product of digestion filed through 80-mesh sieve, and single cells were collected by centrifugation in a density gradient according to Picollo. Cells at the interface were washed and incubated in saline phosphate buffer (PBS) containing 2% FBS (PBS-FBS), 5 μg/ml of immunoglobulin IgG control mice (Coulter Immunology, Hayali, Florida, USA) at a concentration of 106/ml at a temperature of 0oC for 30 minutes. The cells three times washed, and incubated with magnetic beads conjugated with anti-mouse goat IgG (Advanced Magnetics, Cambridge, Massachusetts, USA). The magnetic beads were separated and washed three times by PBS-FBS. This preliminary selection using mouse IgG (mIgG) and magnetic beads used to control non - specific adsorption of T-cells. Remaining in the initial suspension cells additionally incubated with 5 μg/ml monoclonal mouse IgG interacting with IL2-R T-cells at a temperature of 0oC for 30 minutes (Coulter, Immunology, Hayali, Florida, USA). Cells were washed and selected as described above by using magnetic beads. Balls preliminary adsorption of IgG and selected antibodies IL2-R immediately resuspendable in the acidified mixture guanidine, phenol and chloroform, and RNA was obtained according Homesince and Nao which should accurately reflect the number of T cells in synovial tissue at the time of surgical removal. Only half of the mIgG and balls with IL2-R patient 1012 immediately processed for RNA. The rest were cultured in medium RPMI 1640 with 5% FBS, 20% HL-1 (Ventrex the laboratories Inc. , Portland, USA), 25 mm HEPES, glutamine, antibiotics and as a source of IL-2 20% supernatant LAK (Allegretto and others, Science, N 247, page 718, 1990) for 5 days. RNA was extracted from cultures balls IL2-R (1012IL2.d5), and not from the sample 1012mIgG, because there was no viable cells at the end of the 5-day cultivation.

Clone T cells were isolated from centrifugate of the patient 1008. Available in centrifugate cells were cultured with concentration of 2 to 106/ml in medium containing IL-2 for two weeks. Unattached cells from this culture were cloned using the method of limiting dilution on autologous monolayers synovial cells. Got the clone 1008.8 CD4+ cells, which was used for cultivation by regular stimulation of autologous synovial monolayers for 3 days in medium containing IL-2, and then were cultured for 4 days in medium containing supernatant LAK.

Lysis synovial attached to 1008.8 cells

Lysis synovial attached cells 1008.8 displayed the Tr. 219, 1989) using35S as a marker in the experiments CTL. Cells were treated with trypsin, washed and put on the bottom microtitre plate with 96 recesses, and each recess has filed 2000 cells. Cells 1008.8, cultured for 3 days before the study together with synovial attached cells and containing supernatant medium was added to the markers with the following ratio of effectors and markers. Cultures were incubated at temperature 37oC overnight, centrifuged at 300 g for 2 minutes and the radioactivity was determined in 50 µl of the supernatant liquid. Specific lysis was calculated using known formulas based lysed in surface-active substance markers. This clone is a cytotoxic relatively synovial attached cells in experiments CTL (see tab. VII at the end of the description).

Gene amplification-threads receptor T cells by chain reaction using a polymerase

Genes threads receptor T-cells amplified with examples, in various combinations, are given in table. VIII (see the end of the description). V16-dimensional example is degenerated V sample (n = 256), which presumably is associated with 85% of the amplification-threads receptor T cells more than 25 different clones, lines or drugs primary human tissues. Determined the duration of many different genes V data amplified DNA, despite the significant deviation of the sample relative to some groups of V. Thus, PCR amplification using V16-dimensional example facilitates the study of populations of T cells, genes V which specialist is unknown.

Genes threads receptor T cells amplified by two-step reactions with the pairs of examples in table VIII. RNA was subjected to reverse transcription at a temperature of 42oC 40 pmol using Cext example 12 IPF in the conditions described by HART and others, The Lancet, page 596 (1988). The product of the reaction was diluted with a mixture containing 40 pmol V16-dimensional example, nucleotides and buffer, as described above, but without MgCl2in order to achieve a final concentration of 3.6 mm. Samples were denaturiruet at a temperature of 95oC for 15 minutes, was added 1 unit of heat resistant recombinant DNA polymerase (manufactured by Sites Corporation, Emeryville, California, USA, title: AmpliTaqTM) and was carried out by 20 cycles of PCR. Each cycle consisted of denaturation at a temperature of 95oC for 1 minute, renaturation for 2 minutes and respostaoC and 45oC, and the balance is at a temperature of 50oC. One microgram aliquot of sample result data of stage I reactions were added to 100 μg amplificating stage II (see Sites, Gene Amp KitIM) containing 100 pmol Cint example, and 100 pmol of examples V8,V17 or 5C or 100 pmol of examples V dimensional example. The amplification stage II was carried out as described above at a temperature of renaturation 50oC and without treatment at a temperature of 37oC and 45oC.

Samples of RNA from cultures 1012IL2. d5 and 1008.8 amplified using V16-dimensional example and example Cext in the reactions of phase I and using V16-dimensional example and example Cint in the reaction of 35 cycles of phase II. Product reactions with glass beads (Biola, San Diego, CA, USA) was purified from agarose gel with a low melting point, its Foundation was gratuitiously, and its sequence was determined by T7 polymerase (Sequenase, United States Biochem, Cleveland, Ohio, USA) using Cseq example. Numerically predominant sequence V, the corresponding individual rearrangement V17 (see table IX), in a sample 1012IL2.d5 easily moved to reading. Other, less common rearrangement was weak, not readable background stripes in g is away IL2 environment without adding additional cells or antigen does not re-activation of T cells. Thus, the predominance of a single rearrangement V17 in this sample reflects clonal expansion in vivo containing V17 T cells in this patient. Sequencing DNA strands receptor T-cells amplified with the use of cytotoxic clone of T cells, 1008.8, also indicates a rearrangement V17 (see tab. IX at the end of the description). The presence of these two types of samples of synovial T-cells derived from two patients with rheumatoid arthritis, involves containing V17 T cells in the pathogenesis of rheumatoid arthritis.

The presence of rearrangements V17 in other samples of synovial RNA was determined by amplification by PCR using specific regarding V17 example (see table VIII). DNA containing V17 receptor T-cells amplified with the use of samples with magnetic beads obtained from seven patients with rheumatoid arthritis. Using processing products electrophorese the ethidium bromide was found a large amplification V17 in four of the samples containing IL2-R than in the corresponding control samples containing mIgG. This enrichment was not the result of the allocation process, since the amplification containing V8 receptor T cells were not found RA is tov, containing RNA IL2-R, amplified by examples V17 and Cint, and the sequence of the product of the reaction was determined using the example of the Cseq. Samples 1014 and 1015 contained a separate sequence (see table IX), which, like the sample 1012IL2. d5 indicate clonal expansion containing V17 T cells in vivo. In contrast, direct sequencing rearrangements amplified using specific relative V8 example, was not possible due to considerable heterogeneity-filament product.

V17 has the amino acid sequence of MSNQVLCCVVLCFLGANTVDGGI - TQSPKYLFRKEGQNVTLSCEQNLNHDAMYWYRQDPGQGLRLIYYSQIVNDFQ - KGDIAEGYSVSREKKESFPLTVTSAQKNPTAFYLCASS.

HLA-DR-DNA analysis of patients with rheumatoid arthritis

HLA-DR-DNA analysis of patients with rheumatoid arthritis was carried out as follows. DNA from each patient was prepared by boiling 105synovial cells in 200 ál dH2O. 10 μl amplified in 35 cycles in 100 μl of medium (Situs, gene Amp kitTHEM) containing 100 pmol of each example to implement a chain reaction of DR using the polymerase (see table X). 1/10 ál of this environment was reamplification 10 cycles of 10 µl, containing only an example DR2 and 17 pmol 32P-dCTP as the sole source dCTP other (J. Immunol., N 138 p. 1947, 1987), was hybridisable obtaining traces containing 10 pmol of HLA-DR allele-specific oligo (positive fibers). Traces twice washed with chloride of Tetramethylammonium (wood and others, Proc. Natl. Acad. Sd USA, No. 82, page 1585, 1985) at a temperature of 65 - 68oC for 20 minutes, and then treated them with x-rays.

Each patient in this analysis had at least one allele of the genes HLA-DR, namely DR4w4, DR1, DR4w14 or DR4w15 known as causing susceptibility to rheumatoid arthritis factors (see table X in the end of the description).

Containing V17 receptors of T cells or their fragments, which are immunogenic or you can give immunogenicity, can be used for immunization of people using the methods described in example VII. Such immunization may lead to an effective immune response.

Although this invention is described in one preferred embodiments thereof, it is necessary to indicate that various modifications without departing from the scope of the inventive concept. Therefore, the invention is not limited to those disclosed in the text description, but only by the claims.

1. Vaccine for the prevention or treatment rheumatoid fact, as the active substance it contains the receptor of T cells or a fragment corresponding to the receptor of the T cells present on the surface of mediating rheumatoid arthritis T cells, or antiidiotypic antibodies, which is the internal image of this receptor or its above-mentioned fragment, and an active substance taken in immunogene effective amount.

2. The vaccine under item 1, characterized in that the said receptor T-cell contains an amino acid sequence MSNQVLCCVVLCFLGANTVDGGITQSPKYLFRKEGQNVTLSCEQNLNHDAMYWYRQDPGQGLRLIYYSQIVNDFQKGDIAEGYSVSREKKESFPLTVTSAQKNPTAFYLCASS, its modifications not affecting the ability of the receptor to act as an immunogen and immunogene active fragments.

3. The vaccine under item 1, characterized in that said fragment contains the sequence of the variable segment of the above-mentioned receptor of T cells.

4. The vaccine under item 3, characterized in that the above-mentioned sequence of the variable segment is a variable plot-threads.

5. Vaccine for p. 4, characterized in that the said verebelyi plot-thread contains mainly the amino acid sequence of MSNQVLCCVVLCFLGANTVDGGITQSPKYLFRkegqnvtlsceqnlnhdamywyrqdpgqglrliyysqivndfqkgdiaegysvsrekkesfpltvtsaqknptafylcass fragments.

6. Vaccine for p. 5, characterized in that the said variable plot-threads mainly contains sequence SQIVNDFQK.

7. The vaccine under item 1, characterized in that said fragment contains the sequence V(D)j

8. The vaccine under item 1, characterized in that said fragment contains the sequence of the binding site.

9. The vaccine under item 1, characterized in that it further contains an adjuvant.

10. The vaccine under item 1, characterized in that it contains more than one type of receptor T cells or fragments.

11. The vaccine under item 1, characterized in that it contains more than one fragment, corresponding to different sequences of the same receptor of T cells.

12. The vaccine under item 1, characterized in that said fragment is associated with the carrier by conjugation.

13. A method of obtaining a vaccine for the treatment of rheumatoid arthritis, characterized in that you get clones of T-cells that cause the condition, determine the amino acid sequence of the receptor of T cells associated with a condition selected data segments receptors T-cell-specific receptors of T cells, but not for rezeptesammlungen reaction with respect to the receptor of T cells, and thus obtained active ingredient with pharmaceutically tolerable environment implicit known methods.

14. The method of detecting the susceptibility of the individual to rheumatoid arthritis, characterized in that determine T-cell containing variable plot-threads MSNQVLCCVVLCFLGANTVDGGITQSPKYLFRKEGQNVTLSCEQNLNHDAMYWYRQDPGQGLRLIYYSQIVNDFQKGDIAEGYSVSREKKESFPLTVTSAQKNPTAFYLCASS, or its fragment in a sample suffering from rheumatoid arthritis of an individual, which is generally not found in not suffering from rheumatoid arthritis individual, and abnormal expression of T-cell containing such a plot, point to rheumatoid arthritis or susceptibility to rheumatoid arthritis.

15. The method according to p. 14, characterized in that the test is taken from synovial tissue.

16. The method according to p. 14, characterized in that the said variable plot-thread or mentioned sequence is determined using the detector ligand.

17. The method according to p. 14, characterized in that the presence of the abovementioned variable plot-thread or mentioned sequence set using the nucleotide sample, the coding above section or referred to the sequence.

18. Method of prevention and treatment of the containing sequence MSNQVLCCVVLCFLGANTVDGGITQSPKYLFRKEGQNVTLSCEQNLNHDAMYWYRQDPGQGLRLIYYSQIVNDFQKGDIAEGYSVSREKKESFPLTVTSAQKNPTAFYLCASS, with his partner by stimulation of the immune response to receptor T cells, or binding ligand sequence MSNQVLCCVVLCFLGANTVDGGITQSPKYLFRKEGQNVTLSCEQNLNHDAMYWYRQDPGQGLRLIYYSQIVNDFQKGDIAEGYSVSREKKESFPLTVTSAQKNPTAFYLCASS, or with a partner receptor of T cells.

19. The method according to p. 18, characterized in that the partner is a HLA-DR, which causes susceptibility to rheumatoid arthritis.

20. A method of preventing or treating rheumatoid arthritis in an individual, wherein the individual give the agent that is cytotoxic or cytostatic way associated with T-cell containing a sequence SNQVLCCVVLCFLGANTVDGGITQSPKYLFRKEGQNVTLSCEQNLNHDAMYWYRQDPGQGLRLIYYSQIVNDFQKGDIAEGYSVSREKKESFPLTVTSAQKNPTAFYLCASS, or individual induce cytotoxic or cytotoxic immune response against T-cells, containing a sequence MSNQVLCCVVLCFLGANTVDGGITQSPKYLFRKEGQNVTLSCEQNLNHDAMYWYRQDPGQGLRLIYYSQIVNDFQKGDIAEGYSVSREKKESFPLTVTSAQKNPTAFYLCASS.

21. The method according to p. 20, characterized in that the said agent is an antibody bound with a substance from the group, including radioactive substances, chemotherapeutic and hemotoksicheskie substances.

Priority points and features:

21.03.89 p. 1 part of the active substance receptor T cells or its fragment match is also in part of the active substance, specified in paragraph 1;

18.07.89 p. 1 part of the active substance concerning antiidiotypic antibodies, which is the internal image receptor T cells or its fragment, corresponding to a receptor of T-cells found on the surface of mediating rheumatoid arthritis T cells, GD.3, 4, 7, 9 - 12 in the part specified in paragraph 1 of the active substance and p. 8 related to the vaccine, containing as active substance a fragment of the receptor of T cells.

 

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