Treatment and diagnosis of mental disorders

Abstract

To provide a method of treating mental disorders.SOLUTION: Testis development related protein (TDRP; also called Immuno-moodulin or Imood herein) is a circulatory anxiogenic factor that modulates anxiety-like behaviour in mammalian models through the regulation of the immune system. Provided is a method of treating mental disorders comprising administering a TDRP antagonist to an individual in need thereof.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to methods and compounds for the treatment and diagnosis of psychiatric disorders, such as anxiety disorders. [Background technology]

[0002] The difficult life of patients diagnosed with autoimmune diseases is due to the occurrence of psychiatric disorders as a major comorbidity It is often made worse by 1,2 For example, approximately 40% of patients with multiple sclerosis Attempting suicide 3,4 , Over 30% of patients with autoimmune hepatitis suffer from schizophrenia 5 ,6 Most notably, immunomodulatory therapies for the treatment of autoimmune conditions have been shown to address these issues. May exacerbate the outbreak 7 , presenting a paradoxical dilemma to both clinicians and patients. The physical symptoms of autoimmunity, even when effectively alleviated, come at the cost of a worsening emotional state and well-being. For example, interferon-α, which is currently used as an effective treatment for multiple sclerosis, Interferon-β (IFN-β) may be used, but its use is associated with autoimmune disease in a significant proportion of patients. Limited by increased expression of homicidal ideation 8 Functional crosstalk between the brain and immune system has been investigated in several studies 9,10 , how one system affects the other Is there a common root or determinant for the development of psychiatric disorders in autoimmune pathologies? It is not yet clear whether this will be the case.

[0003] Annexin-A1 (AnxA1) is a marker of inflammation 11~14 from autoimmune 15,16 and cancer 1 7~20They are endogenous modulators of a variety of physiological and pathological processes ranging from Like other multifunctional mediators, AnxA1 exerts both positive and negative functions depending on the context. They play a homeostatic role in the immune system, as they can contribute to the maintenance of immune responses. Indeed, AnxA1 is a positive modulator of T cell activation. 21~30 and negative modulation Tar 30~33 Studies have produced contrasting results showing that it can act both as a All of these studies have focused on exogenously administered recombinant AnxA1 (or its mimetics). ) or in AnxA1-deficient mice, in which this protein is absent in all immune cells. It is being done. Summary of the Invention

[0004] We demonstrate that testis development-related proteins regulate anxiety-like behavior in mammalian models. Testis development related protein (TDRP) is recognized as a factor that induces circulatory anxiety. TDRP (referred to herein as Immuno-moodulin or Imood) (also referred to as 'anti-psychotic') are useful, for example, as therapeutic targets in the treatment of psychiatric disorders or for the diagnosis of psychiatric disorders, They may be useful as biomarkers in prognosis, monitoring or assessment.

[0005] A first aspect of the present invention is a method for treating a psychiatric disorder, comprising administering a TDRP antagonist to a subject. and administering to an individual in need thereof.

[0006] A second aspect of the present invention relates to a TDRP antagonist for use in a method according to the first aspect. provide.

[0007] A third aspect of the present invention relates to the use of TDR in the manufacture of a medicament for use in the method according to the first aspect. The present invention provides the use of P antagonists.

[0008] A fourth aspect of the present invention relates to a method for treating a rheumatoid arthritis (HAT) disease comprising administering to a patient a therapeutically effective amount of a TDRP antagonist and a pharmaceutically acceptable salt thereof. and an excipient.

[0009] The pharmaceutical composition of the fourth aspect of the present invention is a pharmaceutical composition comprising the compound of the first aspect, the compound of the second aspect and the compound of the third aspect of the present invention. It may be useful in

[0010] Suitable TDRP antagonists for use in the first to fourth aspects include antibody molecules. The children are listed and described elsewhere herein.

[0011] A fifth aspect of the present invention provides a method for assessing anxiety in an individual, comprising: The present invention provides methods for detecting TDRP in a mammalian cell, comprising determining the level or amount of TDRP in the mammalian cell.

[0012] A sixth aspect of the present invention provides a method for diagnosing a psychiatric disorder in an individual or for determining risk of suffering from a psychiatric disorder. A method for identifying an individual with a high risk of developing TDRP, comprising measuring the level of TDRP in a sample obtained from the individual. or determining the amount.

[0013] A seventh aspect of the invention is a method of monitoring an individual undergoing treatment, comprising: determining the level or amount of TDRP in a sample obtained from an individual receiving Well, we provide a method.

[0014] The individual may, for example, be undergoing treatment for an immune condition or a psychiatric disorder.

[0015] An eighth aspect of the present invention is a method for screening compounds having therapeutic activity against psychiatric disorders. The method comprises measuring the level or amount of a test compound in a non-human mammal. and determining the effect of the compound on the psychiatric disorder, wherein a reduction in the level or amount of TDRP indicates that the compound is effective against the psychiatric disorder. The present invention provides a method for determining whether a compound has a therapeutic activity.

[0016] Therapeutic activity against psychiatric disorders may include anti-anxiety activity.

[0017] A ninth aspect of the present invention is a method for determining T cell activation in an individual, comprising: determining expression of TDRP in a sample of T cells obtained from the do.

[0018] Other aspects and embodiments of the invention are described in more detail below. [Brief explanation of the drawings]

[0019]

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Figure 7

[0020] The present invention provides a novel mechanism for the identification of TDRP in T cells, which regulates emotional and anxiety behaviors in mammals. TDRP is a neurotransmitter that is released by the nervous system and is an anxiogenic factor. Biomarkers for the diagnosis and prognosis of psychiatric disorders, and targets for interventions in the treatment of psychiatric disorders It can be useful as a target.

[0021] Testis development-related protein (TDRP) (Gene ID number: 157695; 2610 019F03Rik; also as Inm01, TDRP1, TDRP2 and C8orf42 known as "Immuno-moodulin" or "Imood" herein. The TDRP (as defined above) can be a mouse or, more preferably, a human TDRP.

[0022] Human TDRP is identified in the database accession number NP_001243042.1 ( sequence number 1), the amino acid sequence of NP_778250.2 (SEQ ID NO: 2), or any of these TDRP can have one of the mutations listed in the database accession number NM_00 1256113.1, NM_175075.4, or any of these The polypeptide may be encoded by any variant thereof, such as an isoform or an allelic variant.

[0023] Mouse TDRP is identified by database accession number AAI45287.1 (SEQ ID NO: 17) or a variant thereof. Mouse TDRP can be found in the database the nucleotide sequence of accession number NM_001361625.1, or a variant thereof; It can be encoded, for example, by an isoform or an allelic variant.

[0024] A variant of a reference TDRP amino acid or nucleotide sequence is a variant of the reference amino acid or nucleotide sequence. At least 80%, at least 85%, at least 90%, at least 90% The sequences may have 5% or at least 98% sequence identity. Sequence identity is generally , algorithm GAP (GCG Wisconsin Package™), Acceler ys, San Diego CA). The two complete sequences were then combined using the scrub algorithm (J. Mol. Biol. (48): 444-453 (1970)). Aligns to maximize the number of matches and minimize the number of gaps. Generally, the default with the parameters: gap creation penalty = 12 and gap extension penalty = 4 Although using GAP may be preferable, the default parameters are generally used. Other algorithms that are used include BLAST or TBLASTN (Altschul et al. (1990) J. Mol. Biol. 215: 405-410), FASTA (using the method of Pearson and Lipman (1 988) PNAS USA 85: 2444-2448), or by using the Smith-Waterman algorithm. Rhythm (Smith and Waterman (1981) J. Mol. Biol. 147: 195-197) may also be used.

[0025] Specific amino acid sequence variants include 1 amino acid, 2 amino acids, 3 amino acids, 4 amino acids, 5 amino acids, 6 amino acids, 7 amino acids, 8 amino acids, 9 amino acids, 10 amino acids, 11 amino acids, 12 amino acids, 13 amino acids, 14 amino acids, 15 amino acids, 16 amino acids, 17 amino acids, 18 amino acids, 19 amino acids, 20 amino acids, 21 amino acids, 2 , 8, 9, 10, 10 to 20, or 20 to 30 amino acid insertions or additions, It may differ from the given sequence by substitutions or deletions. In some embodiments, the variant sequence is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more The reference sequence may contain up to 15 inserted, deleted, or substituted residues. , insertions, deletions of up to 20, up to 30, up to 40, up to 50, or up to 60 residues It may be missing or substituted.

[0026] In some embodiments, TDRP is a biomarker for the diagnosis or prognosis of a psychiatric disorder. For example, the detection of TDRP protein in a sample obtained from an individual may be useful as a target. or the level or amount of the encoding nucleic acid may provide diagnostic or prognostic information regarding a psychiatric disorder in an individual. may provide post-diagnostic information or may be an indicator of an individual's emotional or anxiety state. These factors may be predictive.

[0027] In some embodiments, the level or amount of TDRP in a sample from an individual is For example, an increase in the level or amount of TDRP may be an indicator of the severity of a psychiatric disorder in an individual. Increased levels of TDRP may be an indicator of increased severity. For example, TDRP levels of 1 ng / ml to 5 ng / ml Plasma concentrations of 50ng / ml to 80ng / ml of TDR are indicative of mild psychiatric disorders. Plasma concentrations of P can be an indicator of severe psychiatric disorders.

[0028] In another embodiment, the level or amount of TDRP in a sample derived from an individual is determined by measuring the level or amount of TDRP in the individual. This may suggest or diagnose the type of mental disorder present.

[0029] (i) diagnosing or prognosing a mental disorder in an individual, or (ii) suffering from a mental disorder (iii) identifying individuals at risk or (iv) determining an individual's anxiety level. This may involve determining the presence, level or amount of TDRP in a sample obtained from the body.

[0030] In some embodiments, the level or amount of TDRP in a sample obtained from an individual is determined. A suitable control is the level of TDRP in a control sample. or a control sample set obtained from a healthy population not suffering from a psychiatric disorder. A suitable control sample may comprise the average level or amount of TDRP in a subject suffering from a psychiatric disorder. (i.e., not diagnosed with a mental disorder), age-matched, and sex-matched. It can be obtained from healthy volunteers.

[0031] The level or amount of TDRP in a sample obtained from an individual is increased compared to a control. is whether an individual suffers from a psychiatric disorder, is at high risk for a psychiatric disorder, or has a high level of anxiety This could be an indicator that

[0032] In other embodiments, the level or amount of a TDRP is measured over a period of time, e.g., a day, a week, or It can be determined in samples obtained from an individual over a period of one month. Increased levels or amounts of DRPs, e.g., increased levels or amounts of TDRPs in a sample obtained from an individual An increase in the amount of α-glucan at the second time point compared to the first time point indicates that the individual is suffering from a psychiatric disorder. or may be an indicator of increased risk of mental disorders or elevated levels of anxiety. obtain.

[0033] Suitable samples for use according to the present method include serum, plasma, lymph, leukocytes, blood, blood fractions, and the like. Examples of samples include urine, synovial fluid, spinal fluid, saliva, mucus, tears, and sweat. In some cases, plasma, serum, saliva or urine may be preferred.

[0034] In some embodiments, the sample is peripheral blood mononuclear cells (PBMCs) from an individual. The blood cells may include blood cells such as:

[0035] The presence, level or amount of TDRP in a sample can be determined by any convenient means. and many suitable techniques are known in the art.

[0036] The mode of determining binding is not a feature of the present invention and is left to the discretion of those skilled in the art according to their own preference and general knowledge. A suitable method for determining the presence or amount of TDRP as described above can be selected depending on the circumstances. Possible approaches include protein-based methods, such as Western blotting and immunoassay. immunofluorescence, enzyme-linked immunosorbent assay (ELISA), mass spectrometry (MS), radioimmunoassay (RIA), immunoradiometric assay (IRMA), fluorescence activated cell sorting (FACS), flow -cytometry (FC), mass cytometry (CyTOF) and immunoenzymatic analysis (IE) MA) (Sandwich assay using monoclonal and / or polyclonal antibodies) and nucleic acid-based methods such as Northern blotting, RT-PCR, and and microarray analysis. All of these approaches are known in the art. In some preferred embodiments, ELISA, FACS, or RT-PCR are used. It can be done.

[0037] In some embodiments, a sample obtained from an individual is contacted with a recognition agent, such as an antibody molecule. The binding of TDRP in the sample to the recognition agent can then be determined.

[0038] Any suitable means can be used to determine binding of the recognition agent. One possibility is to tag the reporter molecule with a detectable, preferably can directly or indirectly generate a measurable signal. The bond may be covalent, directly or indirectly, for example via a peptide bond, or non-covalently. Linkage via a peptide bond may be used to link the antibody and the reporter molecule. For example, a recognition agent, such as an antibody, can be the result of recombinant expression of a gene fusion that recognizes the FIT fluorophores such as C or rhodamine, radioisotopes, or biotin or digoxin The recognition agent may be labeled with a non-isotopic labeling reagent such as sigenin, and may contain biotin. Using a "detection reagent" such as avidin conjugated to any desired label such as a fluorescent dye Another possibility is to use a second recognition agent, e.g. in an immunoassay system. The goal is to detect binding of the recognition agent to TDRP using a method similar to that described above. Depending on the assay format used, Optionally, the second recognition agent can be immobilized or labeled with a detectable label.

[0039] The recognition agent is preferably an antibody molecule. The presence or amount of TDRP in a sample can be determined by, for example, Determined by measuring the formation of immune complexes between antibody molecules and TDRP in the sample The presence or degree of binding can be determined, for example, by agglutination, or a color change or fluorescence, etc. A visible change in, for example, immunostaining, or radioimmunological or enzyme-linked immunosorbent assay This can be shown by quantitative methods such as the use of

[0040] In some preferred embodiments, the presence of TDRP in a sample is detected using an immunoassay. Examples of immunoassays include antibody capture assays, double antibody sub-assays, and the like. These are sandwich assays, ELISA assays, and antigen capture assays. In immunoassays, two antibodies capable of binding to TDRP are generally used, e.g. For example, one molecule may be immobilized on a solid support and the other may be free in solution and labeled with a detectable chemical compound. Examples of chemical labels that can be used on the second antibody include radioisotopes, fluorescent compounds, and the like. , spin labels, colored particles such as colloidal gold and colored latex, and reactants or enzyme substrates. an enzyme or other molecule that produces a colored or electrochemically active product when exposed to When a sample containing TDRP is placed in this system, TDRP is immobilized. The antibody binds to both the immobilized antibody and the labeled antibody, forming a "sandwich" immune complex on the surface of the support. Unbound sample components and excess labeled antibody are washed away, and the antibody is then bound to the surface of the support. The amount of labeled antibody complexed with the protein is measured to determine the amount of complexed protein. Alternatively, the protein can be labeled with a chemical moiety, e.g., a hapten. Detectable binding of free antibody in solution to free antibody or, for example, a hapten linked thereto The immunoassay can be detected by a third antibody labeled with a suitable moiety. Preferably, the immunoassay is a ELISA-based immunoassay. Alternatively, the immunoassay may be any of the methods known in the art. One of the well-known immunoprecipitation methods, such as nephelometric immunoassay or turbidimetric immunoassay When Western blot analysis or immunoassay is used, conjugated enzyme labels can be used. It is preferable to include the method.

[0041] The recognition agent is conveniently an antibody, although other recognition agents may be known or become available. For example, antibodies such as Fab fragments can be used in the present invention. The antigen-binding domain fragment of the antibody can be used. Therefore, unless the context specifically dictates otherwise, The term "antibody" as used herein is intended to include other recognition agents. Optionally, the antibody may be polyclonal or monoclonal. generating antibodies by the method to recognize preselected epitopes derived from can be done.

[0042] In another embodiment, the presence or amount of a nucleic acid encoding a TDRP is determined in a sample. Suitable methods are known in the art.

[0043] In some embodiments, the individual has a disease state, such as an immune condition, an immune-related condition, or a psychiatric disorder. may have

[0044] The individual may be undergoing treatment for a disease condition. For example, the methods described herein include: May be useful in determining the onset or risk of onset of a psychiatric disorder in patients undergoing treatment The method for monitoring an individual receiving treatment involves measuring the level of a protein obtained from the individual receiving treatment. This may include determining the level or amount of TDRP in the sample.

[0045] The individual may be undergoing treatment for an immune condition, such as an autoimmune disorder. Anti-inflammatory drugs, such as nonsteroidal anti-inflammatory drugs (NSAIDs) or immunosuppressants, such as IFN-β , corticosteroids, mTOR inhibitors, calcineurin inhibitors, Janus kinase inhibitors agents, IMDH inhibitors, anti-CD3 antibodies, anti-CD25 antibodies, anti-TNF antibodies, e.g., adalimumab and anti-TNF receptor agonists, such as infliximab.

[0046] Autoimmune disorders, such as T cell-dependent autoimmune diseases such as multiple sclerosis, autoimmune hepatitis, I Type 2 diabetes, celiac disease, Graves' disease, inflammatory bowel disease (IBD), psoriasis, rheumatoid arthritis , systemic lupus erythematosus, myasthenia gravis, idiopathic thrombocytopenic purpura, ankylosing spondylitis, and For example, an individual may be receiving IFN-β treatment for multiple sclerosis. may be receiving medical treatment.

[0047] The individual may be undergoing treatment for an immune-related condition. The immune-related condition may be atherosclerosis. These may include dementia, Alzheimer's disease, and Parkinson's disease.

[0048] The individual may be undergoing treatment for a psychiatric disorder, such as an anxiety disorder. For example, the individual may be receiving antipsychotics, anxiolytics, tranquilizers, mood stabilizers, or or antidepressants, or non-steroidal antidepressants such as talking therapies, cognitive behavioral therapy (CBT) or relaxation techniques. Treatment with medication may also be undertaken.

[0049] In some embodiments, in a sample obtained from an individual with an autoimmune disorder The presence of an elevated level or amount of TDRP compared to a control indicates that an individual has an autoimmune disorder. In addition to treating autoimmune disorders, e.g. For example, the treatments described above, or the TDRP antagonists described herein, e.g., anti-TDRP Antibodies such as 1B10 or 1C4 may be used to treat an individual with a psychiatric disorder. elevated levels in samples obtained from individuals with autoimmune disorders compared to controls or The absence of these amounts of TDRP can be an indication that an individual does not have a psychiatric disorder. For example, an individual may be treated for an autoimmune disorder using the treatments described above.

[0050] Furthermore, the identification of TDRP as a biomarker for psychiatric disorders will be useful for understanding the progression of psychiatric disorders. provides an easily measurable proxy for monitoring progression or regression of a mental disorder or Treatments intended to reduce or eliminate the symptoms should also reduce or eliminate TDRP. be.

[0051] Therefore, a method for assessing the effectiveness of a drug in treating a psychiatric disorder in an individual is For example, the effect of a drug on the expression, level, amount, or concentration of T The expression, level, amount or concentration of the DRP was measured from the individual before and after administration of the drug. The decrease in the expression, level, amount or concentration of TDRP can be determined in the sample. It is an indicator of the effectiveness of drugs in treating mental disorders.

[0052] The increase or decrease in expression, level, amount or concentration of the TDRPs described herein may be a TDR The significance can be a significant increase or decrease in P. Significance can be measured, for example, by Student's t-test or Weinstein's test. This can be measured using a t-test such as the Luci t-test, and a significance level of p<0.001 is significant. In other embodiments, p<0.05, e.g., p<0.01 or In some cases, a significance level of p<0.005 indicates a significant increase or decrease.

[0053] In some embodiments, the increase or decrease in the expression, level, amount, or concentration of TDRP is: The expression, level, amount or concentration of TDRP in a test sample is higher than that of TDRP in a reference or control sample. At least 110%, at least 115%, at least At least 120%, at least 125%, at least 130%, at least 135%, at least At least 140%, at least 145%, at least 150%, at least 155%, at least At least 160%, at least 165%, at least 170%, at least 175%, at least at least 180%, at least 185%, at least 190%, at least 195% or less It can be significant if both are 200%.

[0054] The mental disorders described herein are characterized by distress, incapacity, or loss of life, pain, incapacity, or freedom. It is a clinically significant behavioral or psychiatric condition characterized by an increased risk of loss.

[0055] Psychiatric disorders may include anxiety disorders, depression, suicidal ideation, and schizophrenia. Characterized by feelings of anxiety and fear. Anxiety disorders include obsessive-compulsive disorder (OCD), generalized anxiety disorder, and generalized anxiety disorder. May include agoraphobia, panic disorder, phobias, post-traumatic stress disorder and social anxiety disorder In some embodiments, the psychiatric disorder described herein is attention deficit hyperactivity disorder (ADHD). In some embodiments, the cognitive impairment described herein may not include ADHD. Mental disorders may not include schizophrenia.

[0056] The diagnostic criteria for mental disorders, including anxiety disorders, are based on the Diagnostic and Statistical Manual of Mental Disorders, 5th Edition (American Society of Mental Disorders). The DSM-5 (Diagnostic, Medical, and Psychological Disorders; also known as DSM-5) lists the disorder as a mental disorder.

[0057] In other embodiments, TDRP may be a therapeutic target for the treatment of psychiatric disorders. For example, a method for treating a psychiatric disorder in an individual in need thereof may include administering a TDRP antagonist. A related aspect is a TDRP for use in such a method of treatment. antagonists and TDRP antagonists in the manufacture of medicaments for use in such methods of treatment. Provide use of the strike.

[0058] TDRP antagonists inhibit the expression, activity, level, amount, and / or is the concentration, particularly the expression, activity, level, amount and / or concentration of TDRP in the plasma or serum of an individual. is an agent or compound that reduces

[0059] TDRP antagonists inhibit the interaction between TDRP and its receptor, ligand, or binding partner. capable of inhibiting, blocking or disrupting an interaction or into the plasma or serum of an individual The secretion of TDRP can be reduced or inhibited.

[0060] The TDRP antagonist may be used to inhibit the expression of TDRP in an individual, for example, in the individual's blood cells, such as PBMCs. The expression of a DRP can be reduced or the expression of a TDRP in the blood or serum of an individual can be reduced. The level, amount, concentration or activity can be decreased or reduced. The TDRP antagonist may be an inverse agonist of TDRP.

[0061] Suitable TDRP antagonists include small organic molecules, receptors, antibody molecules, aptamers, C Targeted gene editing systems such as RISPR / Cas9, and support systems such as RNAi and antisense Lesser nucleic acids are included.

[0062] In some preferred embodiments, the TDRP antagonist specifically inhibits TDRP. It may be an antibody molecule that binds.

[0063] Antibody molecules may be natural, or partly or wholly synthetically produced. The term refers to any polymer that contains an antibody antigen-binding site. The antibody may be isolated from a natural source or purified. obtained by genetic engineering or by chemical synthesis. and may contain unnatural amino acids.

[0064] An antigen-binding site is the part of a molecule that recognizes and binds to all or part of a target antigen. In an antibody molecule, this is called the antibody antigen-binding site or paratope, which binds the entire target antigen. This includes the portion of an antibody that recognizes and binds to the antigen or part of it. An antibody may bind only to a specific part of an antigen, which is called an epitope. The binding site may be provided by one or more antibody variable domains. An antibody antigen-binding site comprises: It preferably comprises an antibody light chain variable region (VL) and an antibody heavy chain variable region (VH).

[0065] An antigen-binding site can be provided by the arrangement of complementarity-determining regions (CDRs). Generally, a structure carrying a set of CDRs is one in which the CDR or set of CDRs is a rearranged immunoglobulin. CDRs or VL of naturally occurring VH and VL antibody variable domains encoded by globulin genes is an antibody heavy or light chain sequence, or a portion thereof, at positions corresponding to a set of CDRs. The structure and location of immunoglobulin variable domains are described in Kabat et al. (1987) (Sequences of f Proteins of Immunological Interest. 4 th Edition. US Department of Health and H uman Services.), and is currently available on the Internet (at immuno.bme.nwu.edu or Use any search engine to find "Kabat" and refer to the revised version. can be determined.

[0066] The CDR region or CDR is a region of interest that is a sequence of amino acids that are identical to those of the amino acid sequence of the CDR described by Kabat et al. (1987) Sequences of Proteins of Immunol ocular Interest, 4 th Edition, US Department of Health and Human Services(Kabat) et al., (1991a), Sequences of Proteins of Immunological Interest, 5 th Edition, U S Department of Health and Human Services, Public Service, NIH, Washington and its subsidiaries The term "antibody" refers to the hypervariable regions of immunoglobulin heavy and light chains defined by the nucleotide sequence of the nucleotides (later versions of the nucleotide sequence ... immunoglobulin). It is contemplated that antibodies typically contain three heavy chain CDRs and three light chain CDRs. "CDR" The term "CDR" or "CDRs", as the case may be, refers to the antigen or epitope it recognizes. These contain most of the amino acid residues involved in the affinity binding of the antibody to One or several of the regions, or even all of these regions, may be indicated.

[0067] Of the six short CDR sequences, the third CDR (HCDR3) of the heavy chain has a larger sequence. Variability in size (essentially due to the mechanisms of gene arrangement that give rise to it) ) HCDR3 can be as short as 2 amino acids, but the longest known size is 26 Functionally, HCDR3 plays a part in determining antibody specificity ( Segal et al., (1974), PNAS, 71:4298-4302, Amit et al., (1986), Science, 233:747- 753, Chothia et al., (1987), J. Mol. Biol., 196:901-917, Chothia et al., (1989), Nature, 342:877-883, Caton et al., (1990), J. Immunol., 144:1965-1968, Sharon e. t al., (1990a), PNAS, 87:4814-4817, Sharon et al., (1990b), J. Immunol., 144:486 3-4869, Kabat et al., (1991b), J. Immunol., 147:1709-1719).

[0068] Because antibodies can be modified in many ways, the term "antibody" refers to, for example, TDR Any specific antibody having an antigen-binding site with the required specificity and / or binding ability for P Thus, the term should be interpreted to include naturally occurring binding members or substances. any antibody containing an antigen-binding site, whether artificial or wholly or partially synthetic and antibody fragments, particularly antigen-binding fragments and derivatives, containing any polypeptide. Therefore, different polypeptides (e.g., belonging to different antibody classes or subclasses) may be used. Chimeric antibodies include chimeric molecules containing an antibody antigen-binding site or equivalent fused to a The cloning and expression are described in EP 0 120 694 and EP 0 120 695. 25023, and in numerous subsequent publications.

[0069] As mentioned above, fragments of a whole antibody can perform the function of binding to an antigen. Examples of binding fragments include (i) Fab fragments consisting of VL, VH, CL and CH1 domains. (ii) an Fd fragment consisting of the VH and CH1 domains; and (iii) (iv) an Fv fragment consisting of the VL and VH domains of a single antibody; dAb fragments consisting of dAb fragments (Ward et al. (1989) Nature 341, 544-546, McCaffer ty et al., (1990) Nature, 348, 552-554, Holt et al. (2003) Trends in Biotechnolo gy 21, 484-490), (v) isolated CDR regions, and (vi) two linked Fab fragments. (vii) a F(ab')2 fragment, which is a bivalent fragment comprising two domains The VH domains are separated by a peptide linker that allows them to associate to form the antigen-binding site. Single-chain Fv molecules (scFv) in which the VL and VL domains are linked (Bird et al. (1988) Sci ence, 242, 423-426, Huston et al. (1988) PNAS USA, 85, 5879-5883), (viii) Bispecific single-chain Fv dimers (International Application No. PCT / US92 / 09965), (ix) "Diabodies" (national standards) are multivalent or multispecific fragments constructed by gene fusion. International Publication No. 94 / 13804, Holliger et al. (1993a), Proc. Natl. Acad. Sci. USA 90 6444-6448), and (x) each of the VH and VL domains in the set is short or "unusual." The diabody format is a single-chain diabody connected by a "mobile" peptide linker. The V, scFv or diabody molecule comprises a disulfide bridge linking the VH and VL domains. It can be stabilized by incorporating a bridge (Reiter et al. (1996), Nature Bio tech., 14, 1239-1245). Single-chain Fv (scFv) can be prepared, for example, by the method described in (Li et al., (1997), Prot Miniimmunoglobulins or small immunoglobulins as described in (Immunoglobulin Engineering, 10: 731-736) The SIP may be comprised within a homodimeric miniimmunoglobulin protein (SIP). It fuses to the CH4 domain of the human IgE secretory isoform IgE-S2, which forms the human antibody molecule. The combined scFv molecules (ε S2 -CH4;Batista et al., (1996), J. Exp. Med., 184: 2 197-205). A minibody (M) containing an scFv joined to a CH3 domain. It is also possible to generate inibodies (Hu et al. (1996), Cancer Res., 56(13):3055-61 Other examples of binding fragments include those containing one or more cysteines from the antibody hinge region. The addition of a few residues to the carboxyl terminus of the heavy chain CH1 domain results in a Fab fragment. cysteine ​​residue(s) in the Fab' and constant domains that differ from the is a Fab' fragment having a free thiol group, Fab'-SH. The antibody molecule may comprise a human or humanized antibody molecule.

[0070] Suitable antibody molecules are those capable of specifically binding to TDRP, for example in immunoprecipitation experiments. For example, it can reduce anxiety-like behavior in mouse models.

[0071] Antibody molecules that specifically bind to TDRP can be generated using conventional techniques, including, for example, genetic immunization. (See, e.g., Bates et al. Biotechniques (2006) 40(2) 199-208) (See references below.) Suitable proteins for immunization to generate antibody molecules that specifically bind to TDRP. Protein sequences can include SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:17.

[0072] Antibody molecules suitable for use as described herein include 1B10 and Examples include monoclonal antibodies such as 1C4.

[0073] In some embodiments, the 1B10 antibody molecule comprises a VH domain and a VL domain. wherein the VH domain is VHCDR1 of SEQ ID NO: 3 or a variant thereof, SEQ ID NO: 4 or a variant thereof, and VHCDR3 of SEQ ID NO: 5 or a variant thereof. , the VL domain is VLCDR1 of SEQ ID NO: 6 or a mutant thereof, VLCDR2 of SEQ ID NO: 8 or a variant thereof, and VLCDR3 of SEQ ID NO: 10 or a variant thereof. wherein the VL domain of the 1B10 antibody is VLCDR1 of SEQ ID NO: 7 or a variant thereof; VLCDR2 of SEQ ID NO: 9 or a variant thereof, and VLCDR3 of SEQ ID NO: 10 or a variant thereof It may contain variants.

[0074] The antibody molecule is capable of specifically binding to TDRP.

[0075] The VH domain of the 1B10 antibody molecule described herein has the amino acid sequence of SEQ ID NO:11. or variants thereof, or independently one or more, e.g., two or three, in the framework region or the amino acid sequence of SEQ ID NO: 11 having four or more amino acid substitutions, deletions or insertions The substitution may be a conservative substitution. The VH domain may comprise a nucleotide sequence of SEQ ID NO: 12. The nucleic acid sequence may be encoded by a nucleic acid sequence or a variant thereof.

[0076] The VL domain of the 1B10 antibody molecule described herein is selected from the group consisting of SEQ ID NO: 13 and the sequence Amino acid sequence of No. 14, or any variant thereof, or framework region independently one or more, for example, two, three, or four or more amino acid substitutions, deletions, or The amino acid sequence of SEQ ID NO: 13 or SEQ ID NO: 14 may be included with an insertion. The VL domain may be a nucleotide sequence of SEQ ID NO: 15 or SEQ ID NO: 16. The gene may be encoded by a nucleotide sequence, a nucleotide sequence, or a variant of any one of these.

[0077] The 1C4 antibody molecule may comprise a VH domain and a VL domain, VHCDR1 of SEQ ID NO: 18 or a variant thereof, VHCDR2 of SEQ ID NO: 19 or a variant thereof variants, and the VHCDR3 of SEQ ID NO: 20 or variants thereof, wherein the VL domain is VLCDR1 or a variant thereof of SEQ ID NO: 21, VLCDR2 or a variant thereof of SEQ ID NO: 22, and and VLCDR3 of SEQ ID NO: 23 or a variant thereof.

[0078] The antibody molecule is capable of specifically binding to TDRP.

[0079] The VH domain of the 1C4 antibody molecule described herein has the amino acid sequence of SEQ ID NO: 24 or variants thereof, or independently one or more, e.g., two, three or more, amino acids in the framework regions. or the amino acid sequence of SEQ ID NO: 24 having four or more amino acid substitutions, deletions or insertions The substitution may be a conservative substitution. The VH domain may comprise the nucleotide sequence of SEQ ID NO: 25. It may be encoded by a nucleotide sequence or a variant thereof.

[0080] The VL domain of the 1C4 antibody molecule described herein has the amino acid sequence of SEQ ID NO:26 or a variant thereof, or one or more, e.g., two or three, independently in the framework region or the amino acid sequence of SEQ ID NO: 26 having four or more amino acid substitutions, deletions or insertions The VL domain may comprise a sequence of nucleotides of SEQ ID NO: 27. The substitutions may be conservative substitutions. The nucleic acid sequence may be encoded by a nucleic acid sequence of the ...

[0081] The present invention provides a method for the preparation of a CDR, VH, or VL domain sequence, which is a variant of a reference sequence, such as a CDR, VH, or VL domain sequence, as described herein. The proteins described herein may contain one or more amino acid residues or nucleotides that have been altered compared to a reference sequence. For example, 20 or less, preferably 15 or less, and 10 or less , 5 or less, 3 or less, 2 or 1 amino acid residues or nucleotides are different from the reference sequence For example, the variants described herein may have 20 or fewer amino acids, 15 or more amino acids, or more than 20 amino acids. 10 or less, 5 or less, 3 or less, 2 or 1 amino acid residues or nucleotides The variants described herein may include sequences of mutated reference sequences. For example, the variants described herein may include 20 The following amino acid residues are arranged: 15 or less, 10 or less, 5 or less, 3 or less, 2 or 1 Amino acid sequence changed compared to any one of sequence numbers 1 to 9, 11, 14 to 20, and 22 The one or more altered residues may include any of the VH or VL domain sequences described herein. Preferably, the VH or VH domains are within the framework regions of the VH or VH domains described herein. Variants of the VL domain sequences include VHCDRs 1-3 and VLCDRs disclosed herein. The VH or VL domain sequences may comprise a set of 1 to 3. can contain, for example, one, two, three or four amino acid substitutions in the framework regions.

[0082] Amino acid residues in the reference sequence may be inserted, deleted or substituted, preferably with different amino acid residues. It may be altered or mutated by substitution. Such alterations may occur at one or more sites in the encoding nucleic acid. It may be caused by one or more of the above nucleotide additions, insertions, deletions or substitutions.

[0083] A VH or VL domain sequence described herein that is a variant of a reference sequence, such as a VH or VL domain sequence described herein. The proteins or polynucleotides described herein may have at least one amino acid sequence that is identical to the reference amino acid or polynucleotide sequence. at least 50% sequence identity, at least 55%, at least 60%, at least 65%, At least 70%, at least about 80%, at least 90%, at least 95%, or less For example, the sequences described herein may share 98% or at least 99% sequence identity. The variants of the protein to be tested have at least 50% sequence identity with the reference amino acid sequence. amino acid sequence, for example, SEQ ID NO: 1 to 9 or 11, at least 55%, At least 60%, at least 65%, at least 70%, at least about 80%, at least 9 0%, at least 95%, at least 98% or at least 99% sequence identity Variants of the VH or VL domain sequences may include the amino acid sequences disclosed herein. It is preferable to include a set of VHCDRs 1 to 3 and VLCDRs 1 to 3. Sequence variations relative to the reference sequence occur outside the CDRs in the framework regions of the variable domain. It is preferable to do so.

[0084] Sequence identity is generally determined using the algorithm GAP (Wisconsin GCG package ge, Accelerys Inc, San Diego, USA). The Dolman-Wunsch algorithm was used to align two complete sequences and identify matches. Maximize the number of gaps and minimize the number of gaps. Generally, use the default parameters and Gap creation penalty = 12 and gap extension penalty = 4. GAP is preferred. Other algorithms that generally use default parameters, e.g. For example, BLAST (using the method of Altschul et al. (1990) (Altschul et al., 1990)) , FASTA (using the method of Pearson and Lipman (Pearson and Lipman, 1988)), or is the Smith-Waterman algorithm (Smith and Waterman, 1981), or TBLAS The TN program (Altschul et al., 1990, supra) may also be used. In particular, psi-Bl The AST algorithm can be used (Altschul et al., 1997). Similarities were calculated using Genomequest™ software (Gene-IT, Worcester MA USA). ) can also be used to determine the

[0085] Preferably, sequence comparisons are made over the entire length of the relevant sequences described herein.

[0086] The terms "immunoglobulin," "antibody molecule," and "antibody" refer to an antibody that specifically binds to TDRP. Used interchangeably to refer to any protein containing an antibody antigen-binding site that has the ability to bind to an antibody. It can be done.

[0087] Suitable anti-TDRP antibodies include those approved by Professor Fulvio D'Acquisto under the Budapest Treaty. , deposited in the European Collection of Authenticated Cell Cultures (ECACC), B The antibody produced by the hybridoma known as LP-1B10-G2, or Fulv Deposited at ECACC by Prof. io D'Acquisto, BLP-1C4-B7 / F6-D3 Examples include antibodies produced by hybridomas known as

[0088] Suitable individuals for the treatment described above include rodents (e.g., guinea pigs, hamsters, rats, etc.). , mice), murines (e.g., mice), canines (e.g., dogs), felines ( cats), equids (e.g., horses), primates, simians (e.g., monkeys) monkeys (e.g., marmosets, baboons), apes (e.g., gorillas) , chimpanzee, orangutan, gibbon), or human. .

[0089] In some preferred embodiments, the individual is a human. and has traditionally been used as a model to demonstrate therapeutic efficacy in non-human mammals, particularly humans. mammals (e.g., murines, primates, porcines, canines, or Lagomorphs) may be used.

[0090] An individual with a mental disorder is diagnosed with the mental disorder according to clinical criteria known in the art. The individual may exhibit at least one discernible sign, symptom, or laboratory finding sufficient to identify the individual. Examples of clinical criteria can be found in medical textbooks such as the Diagnostic and Statistical Manual of Mental Disorders, 5th Edition. In some embodiments, the individual has previously been identified or diagnosed as having a psychiatric disorder. Alternatively, the methods of the invention may involve the use of a subject that has been obtained from an individual, e.g., as described herein. determining the level or amount of TDRP in the sample obtained to determine the level or amount of TDRP in the individual Identifying or diagnosing the presence of a mental disorder, prognosing a mental disorder, or This may include assessing the risk of developing harm.

[0091] Treatment may involve any number of different conditions, whether human or animal (e.g., in veterinary applications). Any treatment or therapy in which a desired therapeutic effect is achieved, such as inhibiting or slowing the progression of a mental disorder. may be a therapy, reducing the rate of progression, stopping the rate of the mental disorder, or Improvement of one or more symptoms, cure or remission of the mental disorder (whether partial or complete) or to prevent, delay, alleviate or reduce one or more symptoms and / or signs of a mental disorder includes stopping.

[0092] Treatment may include prophylactic treatment (i.e., prophylaxis), i.e., the individual being treated is at risk of developing a prophylactic infection. The individual may not have a mental disorder at the time of the incident or may not have been diagnosed with a mental disorder. For example, individuals susceptible to or at risk of developing or recurring a psychiatric disorder are considered to be individuals described herein. Such treatment may reduce the occurrence of a psychiatric disorder in an individual. or recurrence, or to prevent or delay the onset or recurrence of the symptoms or severity thereof after onset or recurrence. In some embodiments, the individual has a reduced risk of developing autism compared to the general population. may have previously been identified as having an increased susceptibility or risk of a neurological disorder, or may include identifying individuals at increased susceptibility or risk of a mental disorder. Prophylactic or preventative treatment may be preferred in some embodiments. do.

[0093] Although a single TDRP antagonist may be administered, the use of more than one TDRP antagonist is not recommended. The above pharmaceutically acceptable carriers, adjuvants, excipients, diluents, fillers, buffers, stabilizers , preservatives, lubricants, or other materials known to those skilled in the art, and optionally other therapeutic or prophylactic agents. Preferably, the material is provided as a pharmaceutical composition (e.g., a formulation) that also contains the compound. The carrier or other additives must be non-toxic and must not interfere with the efficacy of the active compound. The exact nature of the material may be administered as a bolus, infusion, injection, or any other suitable method, as discussed below. The preferred route will depend on the route of administration. Suitable materials are sterile and pyrogen-free. It has suitable isotonicity and stability. 0.9% NaCl), water, dextrose, glycerol, ethanol, etc., or combinations thereof The composition may also contain auxiliary substances, such as wetting agents, emulsifying agents, pH buffering agents, and the like. That's fine.

[0094] Thus, the methods of the present invention involve the combination of a TDRP antagonist described herein with a pharmaceutical The method may include combining the composition with an acceptable carrier, adjuvant, or excipient. In some embodiments, the TDRP antagonist may be the only active ingredient in the pharmaceutical composition. .

[0095] As used herein, the term "pharmaceutically acceptable" means within the bounds of good medical common sense. within 24 hours without undue toxicity, irritation, allergic reaction, or other problems or complications. Use in contact with tissues of a subject (e.g., human) that is commensurate with a reasonable benefit / risk ratio. The present invention also relates to compounds, materials, compositions, and / or dosage forms suitable for the treatment of various conditions. Each carrier must also be "acceptable" in the sense of being compatible with the other ingredients of the formulation. or other materials, the exact nature of which may be administered, for example by injection, e.g., cutaneous, subcutaneous or intravenous injection. The route of administration depends on the route of administration, which may be parenteral.

[0096] TDRP antagonists can be administered systemically or peripherally, e.g., subcutaneously, intradermally, intramuscularly, or intravenously. Intra-arterial, intracardiac, intrathecal, intraspinal, intra-articular, subcapsular, intraorbital, intraperitoneal, intratracheal, superficial Injections, including subcutaneous, intraarticular, intrathecal and intrasternal, preferably subcutaneous injections; e.g., intradermal, subcutaneous or parenterally by implantation of a depot into the muscle. The TDRP antagonist can be administered to a subject by any convenient route. Preferably, the TDRP antagonist is administered intravenously (IV), intramuscularly (IM), or intradermally (ID). The compound can be administered as a depot injection, for example an intradermal depot.

[0097] For intravenous, cutaneous or subcutaneous injection, or injection into the affected area, The formulation is pyrogen-free and parenterally acceptable with suitable pH, isotonicity and stability. Those skilled in the art will recognize that the formulations of these compounds are in the form of aqueous solutions that are acceptable for various purposes, such as sodium chloride injection, Ringer's solution, or A suitable solution may be prepared using an isotonic vehicle such as lactated Ringer's solution. Preservatives, stabilizers, buffers, antioxidants and / or other additives may be included as required. These may include buffers such as phosphate, citrate and other organic acids; ascorbic acid and methionine; preservatives (octadecyldimethylbenzylammonium chloride) Chloride;Hexamethonium chloride;Benzalkonium chloride;Benzethonium chloride;Phenol butyl alcohol or benzyl alcohol; methylparaben or propylparaben, etc. Alkylparabens; Catechol; Resorcinol; Cyclohexanol; 3'-Penta and m-cresol, etc.); low molecular weight polypeptides; serum albumin, gelatin, or is a protein such as immunoglobulin; a hydrophilic polymer such as polyvinylpyrrolidone; glycine amino acids such as glutamine, asparagine, histidine, arginine or lysine; monosaccharides, disaccharides and other carbohydrates, including saccharide, mannose or dextrin; sugars such as sucrose, mannitol, trehalose or sorbitol; sodium salt-forming counterions such as Zn; metal complexes (e.g., Zn-protein complexes); and / or TWE EN®, PLURONICS® or polyethylene glycol (PE Suitable carriers, excipients, etc. are described in standard pharmaceutical textbooks. Books such as Remington's Pharmaceutical Sciences, 18th edition, Mack Publishing Company pany, Easton, Pa., 1990.

[0098] Pharmaceutical compositions and formulations may conveniently be presented in unit dosage form and are well known in the art of pharmacy. It can be prepared by any known method. Generally, the composition comprises bringing into association the active ingredient(s) with the carrier which constitutes one or more accessory ingredients. The formulations are prepared by uniformly and intimately bringing the compound into contact with a liquid carrier. For example, it may be in the form of a liquid or solution.

[0099] The TDRP antagonist may be administered in a therapeutically effective amount as described herein. A "therapeutically effective amount" is a dose that produces some desired therapeutic effect commensurate with a reasonable benefit / risk ratio. Active compounds, or combinations, materials, and compositions containing active compounds, effective to produce a therapeutic effect The appropriate dosage of the active compound may vary from individual to individual. Determining the appropriate dosage generally depends on the balance between the level of therapeutic benefit and any risk or deleterious side effects of administration. The dose level selected depends on the route of administration, timing of administration, and activation status. the excretion rate of the substance, other drugs, compounds and / or materials used concomitantly, and the individual's age, sex, and physical condition. The severity of your condition may vary depending on a number of factors, including but not limited to your condition, overall health, and medical history. The amount of active compound and the route of administration are ultimately at the discretion of the physician, but generally the dosage is Therapeutic efficacy of the active compound without causing substantial harmful or deleterious side effects The aim is to achieve therapeutic plasma concentrations.

[0100] Generally, a suitable dose of the TDRP antagonist is about 1 kg of body weight per day of the subject. 00 μg to about 400 mg per kg of subject body weight per day, preferably 200 μg to about 20 mg per kg of subject body weight per day 0 mg, for example, in the range of 5 mg / kg / day to 10 mg / kg / day. In the case of esters, prodrugs, etc., the dosage is calculated based on the parent compound. The actual weight used will increase proportionately.

[0101] Pharmaceutical compositions containing the active compound are formulated in dosage unit formulations appropriate for the intended route of administration. It is possible.

[0102] In vivo administration may be in one dose, continuously or intermittently (e.g., at appropriate intervals). This may be achieved in divided doses.

[0103] Methods for determining the most effective means and dosages for administration are known in the art; The composition used in the treatment, the therapeutic purpose, the target cells to be treated, and the subject to be treated will determine the therapeutic effect. Single or multiple doses may be administered depending on the dose level and dosing pattern selected by the physician. A single dose may be administered.

[0104] Multiple doses of the TDRP antagonist may be administered, e.g., two, three, four, five, or more times. The TDRP antagonist may be administered over a prolonged period of time. For example, treatment with a TDRP antagonist can be continued for at least one year. for at least two weeks, at least three weeks, at least one month, or at least two months Treatment with a TDRP antagonist may reduce or eliminate the symptoms of a psychiatric disorder. Treatment may continue for as long as necessary to achieve complete remission.

[0105] TDRP antagonists may be used alone or in combination with other treatments depending on the individual situation. The TDRP antagonist can be administered simultaneously or sequentially. For example, the TDRP antagonist can be administered in combination with one or more with additional active compounds, such as immunomodulators, e.g., immunosuppressants, antipsychotics, or other treatments. Combinations can be administered.

[0106] In other embodiments, the TDRP has therapeutic activity for psychiatric disorders, e.g., anti-anxiety activity. The compounds identified by the screening may be useful for screening compounds having the The compounds may be useful in the development of therapeutics for the treatment of psychiatric disorders.

[0107] A method for screening for compounds having therapeutic activity, e.g., anti-anxiety activity, comprises: and contacting the isolated TDRP with the test compound and / or the isolated TDRP. and determining neutralization of the isolated TDRP by a test compound, Alternatively, neutralization is indicative that the test compound has therapeutic activity for treating the psychiatric disorder.

[0108] In other embodiments, compounds with therapeutic activity against psychiatric disorders, e.g., anti-anxiety activity. The method for screening for TDRP includes screening for the expression, level, amount or The method can include determining the effect of the test compound on the concentration of TDRP, and A decrease in the level, amount or concentration of the compound is indicative of the compound having anxiolytic activity.

[0109] Test compounds identified as having anxiolytic activity may be useful in the development of treatments for psychiatric disorders. obtain.

[0110] Test compounds that can be screened using the methods described herein include drug screening compounds. It can be a natural or synthetic chemical compound used in cleaning programs. Suitable compounds includes TDRP antagonists and variants or derivatives thereof.

[0111] One or more initial screening tests were used to identify compounds that were found to have the ability to bind or neutralize TDRP. Test compounds identified through this process may be further evaluated using one or more secondary screens. Secondary screening can be performed in vitro and / or in vivo, e.g. This may involve testing biological function or activity in animal models. For example, rodent models. The ability of a test compound to reduce anxiety-related behavior in a mouse model can be determined.

[0112] After identifying a test compound that binds or neutralizes TDRP, the compound is isolated and / or purified. or alternatively, the compounds can be synthesized using conventional techniques of chemical synthesis. The pharmaceutical properties of the compound may be optimized using modeling techniques known in the art. Furthermore, the compounds can be manufactured and / or compositions can be prepared using the same method. The compounds can be used in the manufacture or formulation of pharmaceuticals for the treatment of psychiatric disorders. These compounds may be useful as TDRP antagonists in the development of therapies for:

[0113] The data herein also demonstrate that TDRP expression is upregulated upon T cell activation. Therefore, TDRP may be a useful biomarker for T cell activation. The method for determining activation of T cells in a sample is to measure the expression of TDRP in T cells in the sample. This may include determining.

[0114] Increased expression of TDRP in T cells compared to controls indicates that the T cells are activated. It can be an indicator.

[0115] The sample contains T cells, e.g., CD4 + T cells or CD8 + It may be a sample of T cells.

[0116] In some embodiments, the expression of TDRP in a sample of T cells obtained from an individual is determined by TDRP in a sample of T cells compared to non-activated or resting T cells can be determined. Increased expression of may be indicative of the presence of activated T cells in a sample derived from an individual. .

[0117] In another embodiment, the T cells in the sample expressing high levels of TDRP are High levels of TDRP expression can be determined in resting or non-activated T cells. High levels of TDRP expression may occur in only 20% of T cells in a population. An increase in the amount or percentage of T cells in a sample that expresses a specific TDRP is indicative of an increase in the number or percentage of T cells in the sample. This can be an indicator of activation.

[0118] TDRP expression can be determined using standard techniques. In this study, TDRP expression can be measured using immunological techniques, for example, by combining a sample with an anti-TDRP antibody. In another embodiment, the protein level can be determined by contacting the TDRP expression was measured using Northern blotting or RT-qPCR. This can be decided at A level.

[0119] Determining T cell activation using the TDRPs described herein can be used to, for example, diagnose a disease state. Therefore, it may be useful, for example, to assess the level of immune system activation in an individual.

[0120] Other aspects and embodiments of the present invention are defined by the term "consisting of" The aspects described above with the term "comprising" replaced by Modes and embodiments, and the use of "consisting essentially of" The above-described terms "comprising" are replaced by the words The present invention provides aspects and embodiments.

[0121] This application does not, unless otherwise required, cover all of the above aspects and interoperability of the embodiments described above. Likewise, this application is intended to disclose all combinations of the above unless otherwise required. Unless otherwise specified, all preferred and / or optional features alone or in any combination with any other aspect are expressly excluded. Disclose the match.

[0122] Variations of the above embodiments, further embodiments and variations thereof may be found by reading this disclosure. will be apparent to those skilled in the art and as such are included within the scope of the present invention.

[0123] All literature and sequence database entries referred to herein are the property of their respective owners for all purposes. and are incorporated herein by reference in their entireties.

[0124] As used herein, "and / or" refers to each of the two specified features or components. is understood as a specific disclosure of one or the other, including or excluding the other. For example, "A and / or "A" and "B" refer to (i) A, (ii) B, and (iii) a specific disclosure of each of A and B. and are to be understood as if each were individually set forth herein. [Example]

[0125] experiment Materials and Methods Animals and husbandry Mice were housed in groups of six per cage under specific pathogen-free conditions with free access to food and water. Mice were housed for at least 7 days before testing. Wild-type C57BL / 6 Mice were purchased from Charles River. All experiments were performed during the light phase of the light-dark cycle. The following two studies were carried out: All studies were carried out with Home Office permission and in accordance with the UK Animal (Farm) Regulations 1986. The procedure was carried out in accordance with the (biological treatment) method.

[0126] To monitor transgenic colonies, genomic DNA was extracted from ear clips. Extraction from the pool using the REDExtract N-AMP-XNAT kit (Sigma, UK) and AnxA1 tg The samples were analyzed by PCR using the following specific primers: Forward primer 5'-GTATGGAATCTCTCTTTGCCAAGC-3' ; reverse primer: 5'-ACHGATATGCACATCAGGAGGG-3' (Thermo Scientific, UK). PCR reaction parameters are as follows: Initial denaturation at 94°C for 3 minutes, followed by denaturation at 94°C for 45 seconds and annealing at 60°C for 45 seconds. 30 cycles of ring and extension at 72°C for 15 seconds, followed by a final cycle of 72°C for 7 minutes. Elongation process.

[0127] Flow cytometry analysis Thymocytes and lymphocytes were cultured in 100 μl of FACS buffer (5% fetal bovine serum and 0. The staining was performed in phosphate-buffered saline containing 0.2% NaN2. The antibody used was anti-C D3 phycoerythrin (clone 145-2C11), anti-CD4 fluorescein isothiocyanate cyanate (clone GK 1.5), anti-CD8 Cy5 (clone 53-6.7) (all (available from eBioscience, San Diego, CA, USA). Cells were cultured in appropriate media as previously described. The cells were labeled with a conjugated antibody at a concentration of 0.05% for 1 hour at 4°C. After labeling, the cells were washed and then incubated for 1 hour at 4°C. Results were analyzed using a Scalibur flow cytometer. ) software (Tree Star, Ashland, OR, USA, Oregon Corporation) was used for analysis. Ta.

[0128] T cell activation assay Lymph node T cells (1 × 10 5 cells / 200 μl) were incubated with medium alone. or in 96-well plates with plate-bound anti-CD3 and anti-CD28. Lymph node T cells were stimulated with IgG for 24 hours. Stimulation with plate-bound anti-CD3 and anti-CD28 was performed as indicated. After 16 hours, FACS was performed. PE conjugates diluted in buffer (PBS containing 1% FCS and 0.02% NaN2) were Conjugated anti-CD69 (clone H1.2F3) and FITC-conjugated anti-CD25 ( The cells were stained with IgG (clone PC61.5). Intact cells were analyzed using forward and side scatter. The data was then analyzed using FlowJo™ software (Tree Star, Ashland, OR, USA, Oregon). IL-2 production was measured after 24 or 48 hours of stimulation using a standard E. coli ELISA. Measurements were performed using an LISA kit according to the manufacturer's instructions (eBioscience).

[0129] Intracellular staining and cytometric bead assay Th cell phenotype was studied by intracellular staining. Lymphocytes were treated with MOG 35-55 immune mouse Lymphocytes derived from lymph nodes and spleen (1 0×10 6 cells / ml) were treated with medium alone (CTRL) or anti-CD3 and anti-CD28 antibodies ( 1 μg / ml; plate-bound) or specific antigen MOG 35-55 (100μg / ml) Both were cultured for 72 hours. On day 3, the cells were pelleted and the supernatant was stored at -20°C. The cells were then treated with concanavalin A (ConA, 5 μg / ml; Sigma) to detect protein export. Recirculation was performed for 4 hours in the presence of the transport inhibitor Brefeldin A (1:1000; eBioscience). Mononuclear cells isolated from the spinal cord were instead treated with ConA and Bref after collection. Direct induction with eldin A.

[0130] Cells were pelleted and then stained for CD4 (1:500) for 30 min in 1% PFA. The cells were then fixed for 10 minutes with IFNγ, IL-17, and GM-CSF. and conjugated antibodies against cytokines such as IL-10 (dilution: 1:250). Permeabilization buffer containing 0.1% saponin and 0.09% sodium azide in PBS The staining was performed for 30 minutes in a 5% CO2-containing solution (eBioscience) (see Table 6 for details). Finally, the cells were washed and transferred to a FACS buffer for flow cytometry analysis. The mixture was suspended in a buffer.

[0131] Cytometric Bead Array Cytokine production was measured by a bead-based analytical assay in flow cytometry. Mouse Th1 / Th2 10plex and custom-designed Mouse T The h1 / Th2 / Th17 / Th22 13plex kit (eBioscience) was used. The former is GM-CSF, IFNγ, IL-1α, IL-2, IL-4, IL-5, IL-6 , IL-10, IL-17, and TNFα-conjugated beads, the latter of which is IFN- γ, IL-1α, IL-2, IL-4, IL-5, IL-6, IL-10, IL-13, Antibody-conjugated beads for IL-17, IL-21, IL-22, IL-27, and TNFα To this were added beads conjugated with antibodies against GM-SCF and IL-23. A sample (25 μl of cell culture supernatant) was added to 50 μl of the beads mixture and biotin-conjugated After two washes, the plates were incubated with 50 μl of a mixture of the diluted antibodies for 2 hours. Streptavidin PE conjugated antibody was added (Figure 2.11) and the sample was incubated in the dark for 1 Finally, the samples were washed and stored overnight at 4°C. Standards were prepared and processed simultaneously. Table 2.3 lists the parameters used for the analysis on the BD LSR Fortessa. Show the meter.

[0132] Adoption Purified CD4 + T cells were isolated from male wild-type or AnxA1tg mice (6 weeks old) by the manufacturer. Following the instructions, negative selection was performed (Dynabeads Untouched Mouse CD3 Cells and Dynabeads Untouched Mouse se CD4 Cells;Invitrogen, Invitrogen Life Technologies Ltd, Paisley, Purity was tested by fluorescence-activated cell sorting and was greater than 98%. Blood was collected by intracardiac puncture under anesthesia. Plasma was separated from clotted blood by centrifugation (8 Freshly isolated plasma was obtained by centrifugation at 1000 rpm for 5 minutes and stored at 4°C until injection. and cells resuspended in phosphate-buffered saline (2 × 10 6 (300□l) as a recipe The cells were transplanted into male C57BL / 6 mice (6 weeks old) by intraperitoneal injection.

[0133] histology Intact spinal cords were first fixed in 4% paraformaldehyde for 72 hours and then paraffin-embedded. Before embedding, the specimens were incubated with a decalcifying solution containing EDTA (0.1 mM in PBS) for 14 days. Tissue sections (5 μm) were deparaffinized in xylene and analyzed by hematopoietic analysis in our in-house histology facility. Histological evaluation was performed in various ways depending on the severity of the disease. The study was performed on paraffin-embedded sections sampled at various time points. A minimum of three sections per animal were evaluated. Phase contrast digital images were taken using Image I. Image-Pro (Media Cybernetics, Rockville, MD, USA) analysis software package The images were taken using a camera.

[0134] MOG 35-55 Experimental autoimmune encephalomyelitis induced by Male C57BL / 6 mice were given M OG 35-55 (300 g) was injected intradermally, and the results were obtained on days 0 and 10 as previously reported. Pertussis toxin (PTX) was administered twice on days 1 and 2. Disease severity was graded on a scale of 0 = no neurological signs; 1 = tail weakness; 2 = tail paralysis; 3 = loss of righting reflex (after the mouse was turned over on its back) 0 = unable to stand up), 4 = paralysis of hind legs, 5 = quadriplegia and 6 = death. It was scored on a scale of 6.

[0135] Isolation of leukocytes from the central nervous system The spinal column was incised from the lumbar to cervical regions and washed several times with PBS to remove traces of blood. The spinal cord was extracted by hydraulic pressure into the spinal canal using a 1 ml syringe and a 19-gauge needle. The tissue was mechanically strained using a 70 μm mesh cell strainer (Falcon) in sterile PBS. The mononuclear cells and lymphocytes were separated by pressure. The cells were isolated by density gradient centrifugation at 400 x g for 5 min. The 30% Percoll solution was diluted with 70% Percoll solution. The mixture was carefully layered on top of the col solution at a ratio of 1:2 and centrifuged at 500 × g for 30 minutes. In this density gradient, mononuclear cells settle at the interface between the 30% and 70% Percoll layers. Carefully remove the top fat layer from the centrifuge tube and add approximately 2-3 ml of interface fluid. Purified mononuclear cells were collected and treated with 100 U / ml penicillin and streptomycin and 1 The cells were washed twice with RPMI (Invitrogen) supplemented with 0% FCS.

[0136] Pristane-induced lupus Wild-type or AnxA1tg male mice were injected with sterile pristane (Sigma-Aldrich, St. Louis, MO). A single ip injection of 0.5 ml of 100 mg ... In some experiments, mice were sacrificed after 2 weeks and peritoneal lavage was performed. The effusions were collected in PBS / EDTA and the levels of inflammatory cytokines were measured and analyzed as previously described. The phenotype of the recruited activated T cells was analyzed as shown.

[0137] Digging and marble burying tests The marble burying and burrowing tests were performed as previously described, with some modifications. Briefly, the container was filled with wood chip bedding to a depth of approximately 5 cm and gently pressed down. The mouse was placed in a flat, transparent plastic box (14 cm × 10 cm × 11 cm). The same bedding substrate was used for all mice and flattened after each test. The marbles were placed on the surface in five rows of three marbles each. The time spent digging, the number of digging movements, and the number of glass marbles hidden (up to 2 / 3 of the depth) were measured for 15 minutes. Two trials were conducted, the second 24 hours after the first. went.

[0138] Climbing behavior test Vertical activity and exploratory behavior were assessed using a climbing test. The test was performed as previously described. The procedure was carried out as described above, with some modifications. Briefly, mice were placed one at a time. Then, place the cylindrical climbing mesh on a thin layer of fresh wood chip bedding on the laboratory bench. Each was observed for 5 minutes and recorded. The number of climbing events and the total duration of the climbing behavior were assessed. All four of the mouse's legs must be on the wire mesh, and as soon as one leg touches the platform, This test was conducted in the early evening, when mice are known to be more active. I went there.

[0139] Light and Dark Shuttle Box In this study, exploration activity reflects a combination of risk and risk aversion (44). The station consists of a 45cm x 620cm x 621cm box with two different compartments: It was divided into two parts: one third (15 cm long) painted black with a black lid on top, and the remaining Two-thirds of the container is painted white and has no lid. A 2.5cm x 62.5cm opening divides the container into two compartments. To enable behavioral assessment, one side of the light box was made transparent, and a Additional lighting provided by a 50W lamp placed 45cm above the The test was conducted according to a previously published protocol. The rats were placed in the light compartment with their backs to the opening and allowed to explore the box for 5 minutes. The dependent variable was the duration of stay in the light area. Dwell time, time taken to move into the dark area (entering all four paws) and total number of transitions between compartments The equipment was cleaned after each test.

[0140] Open field activity test The open field test was performed as previously described with some modifications. The open field was a white PVC arena divided into 10 cm × 10 cm squares. The mice were brought into the experimental room 15 minutes before the test. Each mouse was placed in one of the corner squares facing the wall and observed and recorded for 5 minutes. The total number of squares reached, the time taken to first stand up, and the total number of stands up were recorded. The arena was cleaned with water after each trial to reduce and standardize olfactory tracking. A second test was conducted 24 hours after the end of the first test.

[0141] Microarray analysis Total RNA was analyzed using the RNeasy Microarray Tissue Mini Kit Extracted from the brains of wild-type and AnxA1tg mice using Qiagen (West Sussex, UK). For purified CD4+ T cells, use the RNeasy Mini Kit from the same manufacturer. Total RNA was purified using Affymetrix Mouse Immunoprecipitation (UCL Genomics, London, UK). Gene 1.0 ST array chips are loaded using standard Affymetrix protocols. Hybridization was performed using the Chip Fluidics Station 450. ffymetrix GeneChip Scanner(Affymetrix,Santa Clara,C The data were scanned using the Bioconductor package affy. Differentially expressed genes were normalized by the RMA of Bioconductor. The limma package was used to identify the disease, taking into account the false positive rate (adjusted P value 0.05). Gene and sample scoring systems were generated by canonical correspondence analysis. Canonical correspondence analysis is a variant of correspondence analysis that uses the primary data The regression data was then analyzed by correspondence analysis. In this study, the entire data set was analyzed using the "average" wild type and "average" Anx A1 tg The regression was performed on explanatory variables defined as the difference between the Signaling pathway impact analysis is performed using the Bioconductor package SPIA. Using wild-type and AnxA1 tg This was done by comparing the

[0142] Real-time polymerase chain reaction Total RNA was analyzed using the RNeasy Microarray Tissue Mini Kit Whole brains (n ​​= 1 / for each mouse strain) were analyzed using Qiagen (Qiagen) according to the manufacturer's protocol. Extract from 4-6), 2 mg of oligo(dT)15 primer, 10 U of AMV reverse transcription transcriptase, 40 U of RNase inhibitor (all from Promega Corporation, Madison, WI, USA), and and 1.25 mM of each dNTP (Bioline, London, UK) at 421°C for 45 min. Real-time polymerase chain reaction was performed using ABsolute™ QPCR®. OX Mix (Thermo Scientific, Epsom, UK) and fluorescent QuantiTect primers Cycling conditions were set according to the manufacturer's instructions. Sequence-specific fluorescence Signals were analyzed using a 7900HT Fast Real-Time PCR System (Applied The mRNA data were analyzed by a chromatographic method (Digidesign Biosystems, Warrington, Cheshire, UK). Expression levels were calculated after normalization to ceraldehyde 3-phosphate dehydrogenase. The comparative Ct method was used to measure gene transcription in the samples. The results were analyzed using 2□DD. It is expressed as a relative unit based on the calculation of Ct, which is the difference with the endogenous control (glyceraldehyde 3-lysine). of genes normalized to the lowest expressing sample, which was set as 1. Relative amounts are given. Total RNA was also analyzed from PBMCs of OCD subjects and healthy controls using Chomczynsk The RT-PCR reaction was performed according to the method of RevertAi and Sacchi (1987). d H Minus First Strand cDNA Synthesis Ki Relative abundance was determined using an iQ S (Thermo Scientific, Waltham, MA, USA). RT-qPCR using YBR Green Supermix (Hercules, CA, USA) DNA Engine Opticon 2 Continuous Fluo rescence Detection System(MJ Research,Waltham,MA,US A) was assessed. To ensure accurate quantification of the initial target in each PCR reaction, the amplification process Production is defined by examining the cells and assigning a fluorescence threshold above background. The point in the early exponential phase of accumulation was defined as the threshold cycle, or Ct. The difference in numbers was used to quantify the relative amount of PCR target in each tube. Dissociation curves (melting curves) were generated in the range of 0°C to 95°C to evaluate the specificity of the amplified products. The relative expression of the different amplicons was calculated by the delta-delta Ct (ΔΔCt) method and statistically For analysis, the relative expression ratio (2 -ΔΔCt ) All human data were converted to the endogenous reference Normalization was performed against a combination of the genes β-actin and GAPDH.

[0143] Western blotting analysis Lymph node and splenic T cells or purified CD4+ T cells were stimulated as indicated in the figure. After incubation, cells were lysed in ice-cold lysis buffer (1% NP-40, 20 mM T ris(pH7.5), 150mM NaCl, 1mM MgCl2, 1mM EGTA , 0.5 mM PMSF, 1 μM aprotinin, 1 μM leupeptin, 1 μM pepstatin Dissolved in 50 mM NaF, 1 mM NaVO4, 1 mM μ-glycerophosphate The supernatant was collected and stored at -20°C. To immunoprecipitate dulin, 5 μl of rabbit polyclonal anti-2610019F 03Rik antibody (Origene, USA) and 35 μl of Protein A / G Sepharose beads , and stimulated with plate-bound anti-CD3 / CD28 (0.5 μg / ml) for 24 or 48 hours. 1×10 7 The sample was added to 500 μl of culture supernatant obtained from T cells at 100 μg / ml. The cells were incubated overnight at 4°C under continuous rotation and then washed with PBS. The precipitate was denatured in hot 6x sample buffer and then subjected to SDS-12% polyacrylamide gel electrophoresis. The samples were then transferred to a PVDF membrane and deionized using a 5% ethanol solution. A Tris-buffered saline solution containing Tween®-20 with dry milk (T TBS:130mM NaCl;2.68mM KCl;19mM Tris-HCl; Antibodies diluted with 0.001% (v / v) Tween®-20, pH 7.4 Both were incubated overnight at 4°C. Immunoblotting and enhanced chemiluminescence (ECL) Protein visualization was performed using a chromatographic method (Amersham Biosciences, Piscataway, NJ, USA) according to the manufacturer's instructions. I followed the instructions.

[0144] Generation of T cell-specific AnxA1tg mice To generate VACD2 Anx-A1 FLAG transgenic mice, The mouse Anx-A1 gene was extracted, amplified, and tagged with the FLAG epitope. into the TOPO pcDNA3.1 vector for validation of its expression in vitro and finally cloned into the VACD2 vector for T cell-specific expression in mice. Finally, the VACD2 Anx-A1 FLAG construct was modified to Purified for pronuclear microinjection into the mouse genome.

[0145] Subject OCD tertiary outpatient Clinic of the University Department of Psychiatry of M Patients of any gender and age who are undergoing treatment and follow-up at Policlinico Hospital, Iran Twenty outpatients with OCD participated in the study. Diagnosis was based on DSM-5 criteria (SCID 5 research edition). In the case of psychiatric comorbidity, the following was assessed: OCD is the primary disorder that causes the most significant distress and impairment and is the primary cause of treatment seeking. The patient had recent or current alcohol or drug abuse. If you have any medical conditions, including autoimmune diseases, For the same reason, patients were excluded from the study because of the possibility that their lifetime Extensive trauma history (according to DSM-5) and current related psychological stress are also exclusion criteria. The clinical assessment included collection of the following demographic and clinical variables: Gender, age, age at onset, and current pharmacological treatment. In addition, disease severity was assessed by Yale-Brown scale. Patients had to be stable for at least 1 month to be enrolled in the study. Control subjects (n=20) were maintained on the same pharmacological treatment as determined by SCID 5. No established psychiatric diagnosis and a positive family history of major psychiatric disorders in first-degree relatives Not (assessed by family interview for genetic studies), gender, age and ethnicity matched All subjects were volunteers. All consent was given to the use of personal and clinical data and to the inheritance of Written consent to participate in a study that includes blood sampling for genotyping and methylation analysis The study protocol was approved by the local ethics committee.

[0146] result T cell-specific AnxA1 tg Mouse generation and phenotypic characterization Pronuclear injection of VACD2 Anx-A1 FLAG construct in 129 FVB mice T cell-specific transgenic mice were generated by the following method. None of the littermates showed any gross signs of disease. Backcrossing and outcrossing onto the C57BL / 6 background to generate mice with After mating, female litters from one of the two transgenic founders exhibited cannibalism ( It was found that the incidence of maternal cannibalism was unusually high (almost 80%). have previously reported on other autoimmune-prone and anxiety-prone mouse strains, such as DBA / 2J. As reported 34 has been successfully controlled by administering perphenazine during pregnancy. The newborns of this strain were kept in foster care by C5 to avoid colony loss. Despite these efforts, it was not possible to maintain this colony. There were problems with the colony and for this reason we ended it. 1 tg Analysis of the immune repertoire of mice revealed CD4 + also is CD8 + No significant difference was observed in the lineage commitment of the cells. AnxA1 compared to controls tg Selective increase in total cell number (approximately 60) in mouse lymph nodes %) was found in the spleen and thymus, but did not increase.

[0147] AnxA1 tg T cell activation and increased autoimmunity in mice Consistent with our previous report 22,23,25,26 , AnxA1 tg T cells , a lower threshold for upregulation of CD25 and CD69 (Fig. 1A) and anti-CD3+CD28 stimulation The mice showed a clear inflammatory phenotype, as evidenced by increased IL-2 production after stimulation (Fig. 1B). In vivo, AnxA1 tg Mice showed increased expression of AnxA1 compared to wild-type tg Worsening severity of clinical scores and weight loss after disease onset (day 12) in mice MOG was associated with increased inflammatory infiltrates in the spinal cord, as evidenced by increased inflammatory cytokines. 35-55 Inducible Experimental This demonstrated an exacerbated inflammatory response in autoimmune encephalomyelitis (EAE) (Figure 1C). In terms of control and AnxA1 tg No differences were observed between the two mice (Table in Figure 1C). To expand and confirm these findings in other models of autoimmune inflammation, we investigated the effects of AnxA 1 tg Mice were subjected to an experimental model of systemic lupus erythematosus 35 Figure 1D (right panel) As shown in Fig. 1, AnxA1 tg Injection of pristane into mice resulted in effective The mice lost approximately 30% of their initial weight over the same period. Consistent with these results, approximately 30% of AnxA1 was reduced during 35 days of treatment. tg Mouse dies However, control animals showed 100% survival (Fig. 1D; left panel).

[0148] AnxA1 tg Selective accumulation of Th1 / Th17 double-positive cells in inflamed tissues of mice AnxA1 in EAE mice tg To further examine the activation status of T cells, The effector phenotype of these cells at day 16 (disease onset) and day 16 (disease peak) was investigated. This allowed us to determine whether the priming phase occurred in the draining lymph nodes on day 9 or in the spinal cord on day 16. We were able to distinguish the effects of AnxA1 overexpression during differentiation. 36 Day 9 Control and AnxA1 tg Comparison of the draining lymph nodes of mice revealed that the latter group had CD4 + A significant increase in the number of T cells (approximately 85%) was observed, but IL-17 + / IFN-γ + or I L-17 + / GM-CSF + No difference was observed in the percentage of double-positive or single-positive T cells. On day 16, CD4 cells recovered from the spinal cord + The number of T cells was significantly higher compared to controls. xA1 tg in the former compared with the latter, but in this case IFN- gamma + / IL-17 + or GM-CSF + / IL-17 + pathogenic 37 Increase in double-positive T cells The fate mapping reporter of Th17 cells in this autoimmune inflammation model was -Research has shown that these double-positive cells are IL-17 single-positive to IFN-γ single-positive at the site of inflammation. or GM-CSF double-positive T cells. 3 8 Consistent with these findings, AnxA1 tg Mice express IL-17 + Cell Percentage Increased levels of IFN-γ (AnxA1 tg Approximately 10% of cases vs. 18% of controls and GM-CSF (AnxA1 tg Approximately 16% of single positive cells (vs. 20% of controls) The corresponding percentage reduction is shown.

[0149] AnxA1 tgIncreased basal anxiety-like behavior in mice AnxA1 in cages tg Direct observation of mice reveals an increased tendency for compulsive burrowing. The behavioral changes associated with the treatment were clearly evident (Wild-type mice in Video 1 and AnxA1 mice in Video 2). tg Ma To thoroughly measure this increase in anxiety, a series of tests were administered to assess anxiety behavior. The classic test of the marble burying test was used to measure burrowing. 3 9 , AnxA1 tg When applied to the The increase was quantified, approximately doubling the time spent in that activity (Figure 2A). In the box test 40 , AnxA1 tg Mice spent more time in the illuminated area than wild-type mice. The number of crossings between the two compartments was significantly reduced (Fig. 2B). Behavioral differences were observed 41 AnxA1 tg The time it takes for a mouse to make its first climb AnxA1 showed a significant increase in the activity of the serotonin receptor β (SPR) and a significant decrease in the time spent in this activity (Fig. 2C). tg Mau The open field test 42 i) the number of squares traversed or ii) The number of rearing episodes did not differ compared to wild-type animals (Figure 2D ), all of these changes were not secondary to a general impairment of locomotor activity.

[0150] AnxA1 tg Whole-brain genetic fingerprinting revealed increased expression of anxiety-related genes It is clear CD4 + T cells exert a direct influence on genes in the brain 45by a number of mechanisms, including Providing homeostatic control of anxiety-like behavior 43,44 This is AnxA1 tg Also applies to the mouse We wondered whether this might be a problem, so we investigated the gene expression profile of the whole brain using microarray analysis. A comparative analysis of the files was performed. The results showed significant differences in the expression levels of 15 genes. , 8 of which were AnxA1 compared with the whole brain of WT mice tg Regulated in the whole mouse brain Of these, erythroid differentiation regulator 1 (Erdr1) was down-regulated. ) 46 and gamma-aminobutyric acid receptor subunit alpha-2 (Gabra2) 47 etc., several It has been associated with alcoholism and affective disorders, including anxiety. RT-PCR analysis of these genes confirmed these results, with approximately 78% of Erdr1 Downregulation of Gabra2 and approximately three-fold upregulation of Gabra2 were observed. T cells exert persistent regulation of distinct gene sets in the brain without any experimental manipulation. can bring about.

[0151] AnxA1 in C57BL / 6 mice tg CD4 + Adoptive transfer of T cells inhibits anxiety-like behavior Increase CD4 + To determine the causal relationship between AnxA1 overexpression in T cells and the anxiety phenotype, To achieve this, control C57BL / 6 or AnxA1 tg CD4 + T cells in C57BL / We generated chimeric mice adoptively transferred into six hosts. tg CD4 + T cells were given Only mice showed a time-dependent increase in anxiety-like behavior, whereas control C57BL / 6 CD4 + T thin Most interestingly, the initial changes in behavior were observed after transfer. It was measurable at 1 week (data not shown) but became prominent at 3 weeks, and this The continued presence of these cells or the sustained release of factors they may produce may contribute to increased anxiety. It is shown that this is possible.

[0152] To address this issue, purified CD4 + Microanalysis of gene expression profiles of T cells Under resting conditions, AnxA1 tg and control CD4 + T cells and No statistical difference was observed between the anti-CD3 Eight genes were identified that were significantly regulated in anti-CD28 stimulated cells. Among them, AnxA1, interferon-inducible 203 and 2610019F03Rik ( Foci development-related protein (Tdrp) was upregulated. It encodes a small protein of approximately 21 kDa (similar to many cytokines) present in the Since I had been studying the function of T cells, I became interested in the last one and decided to investigate its function. We identified this gene and identified AnxA1 tg hypothesized to be involved in the phenotype of anxiety behavior in mice. They named it Immuno-moodulin (Imood).

[0153] AnxA1 tg CD4 + T cells are able to express a new molecule called immunomoodulin. express high levels of anxiogenic factors The microarray results were then analyzed using a commercially available polyclonal antibody against Imood. CD4 + Initially validated using immunoblotting and FACS intracellular staining of T cells The results showed that AnxA1 tg Imood mRNA and protein in The results consistently showed a significant increase in the expression of both proteins (Figures 3A, 3B, and 3C, respectively). C).

[0154] in resting or activated T cells derived from non-transgenic C57BL / 6 mice We first investigated I mood expression. Under basal conditions, approximately 20% of cells expressed high levels of I Activation of CD4+ T cells by the TCR results in a clear increase in the number of these cells. After induction with signal 1 (anti-CD3) and signal 2 (anti-CD28), These numbers doubled (44%) after 1 h (Fig. 7A). By comparing the expression of Imood in + T cells (Fig. 7B), compared with wild-type, An The percentage of high Imood cells in xA1tg mice (approximately 55% of AnxA1tg) % of the wild type and their median fluorescence intensities (approximately 15% of AnxA1tg). These differences gradually decreased, but the anti-C Approximately 80% of AnxA1tg cells were still observed after activation with D3 / CD28. Imood expressing cells had a median fluorescence intensity of 26, whereas approximately 70% of wild-type cells had a median fluorescence intensity of 26. These results were confirmed at the mRNA level, demonstrating that anti-CD3 / CD28 T Upregulation of Imood mRNA and AnxA1tg compared to wild type after cell activation Increased expression of these proteins was observed under both resting and stimulated conditions (Fig. 7C). These findings were confirmed by Western blot. Cells showed increased levels of intracellular Imood compared to controls (Fig. 3C). From cell supernatants of wild-type and AnxA1tg T cells under both resting and activating conditions Immunoprecipitation of Imood in AnxA1tg T cells revealed increased secretion of this protein. The results showed that the α-glucan-1-phosphate dehydrogenase (α-glucan-1) increased the

[0155] To investigate the possible role of Imood in regulating anxiety behavior, we analyzed this gene The recombinant product was administered to wild-type C57BL / 6 mice. , AnxA1 tg The difference between the WT and WT mice was the greatest. As shown in Figure 3D, the intact protein (r- Mice injected with Imood showed a significant increase in anxiety-like behavior, whereas mice injected with denatured (95°C) Mice injected with protein (d-Imood) or PBS showed no increase. Similar to the results observed in adoptive transfer experiments, this change occurred within a few hours after administration. However, it started to become noticeable on the third day after injection and became extremely noticeable on the seventh day. AnxA1 tg To confirm that the compound is involved in increasing anxiety-like behavior in mice, a commercially available Polyclonal antibodies and control serum were administered to these mice. The serum was also administered to wild-type C57BL / 6 mice to test its effects in control mice. As shown in Fig. 1, polyclonal anti-Imood antibodies were given to AnxA1 tg Mouse only However, the time spent in the light (approximately 249%; p<0.001) and the number of crossings (approximately 194%; p<0.0 01), whereas mice given control serum showed no increase. In contrast, administration of the same antibody in wild-type C57BL / 6 mice resulted in a similar increase in the number of hours in light (approximately This resulted in a significant increase in the number of crossings (approximately 76%; p<0.01) and the number of stenosis (approximately 24%; p<0.05). Therefore, there may be an endogenous level of Imood that can regulate basic anxiety behavior in mice. It was suggested that there is.

[0156] To further verify our hypothesis and increase the specificity of the effect, we used genetic immunization. We generated a monoclonal antibody against Imood. 48 (Figure 6). Consistent with previous data. and the highly selective purified monoclonal anti-Imood antibodies 1B10 or 1C4. A1 tg administration to the IgG control (approximately 123% for 1C4, p <0.01; approximately 178% for 1B10, p<0.001) and time in the light (1C 4, approximately 208%, p<0.01; 1B10, approximately 396%, p<0.00 1) significantly increased AnxA1. tg The phenotype of Rescue to the wild-type mouse phenotype.

[0157] Similar to the results observed with polyclonal antibodies, C 57BL6 control mice spent approximately 75% of their time in the light (1C4, p<0.01; 1 86% for B10, p<0.001) and crossing counts (approximately 41% for 1C4) , p<0.05; for 1B10, approximately 68%, p<0.001). Taken together, these data suggest that Imood is a potential therapeutic agent for treating anxiety behavior in mice. This clearly represents an innovative target for the strategy (Figure 4A).

[0158] Increased expression of Imood in patients with OCD Finally, we investigated Imood expression in humans. To this end, we examined 20 subjects with obsessive-compulsive disorder (OCD). The first cDNA samples from PBMCs obtained from patients diagnosed with HIV and 20 healthy controls were analyzed. As shown in Figure 4B, the Imood The expression level was significantly higher than in the control group (approximately 6-fold).

[0159] A growing body of research suggests that mood disorders are caused by cellular and biochemical events rooted in the immune system. There is support for the assumption that this can be caused by 10,49,50 Mental health The lack of new therapeutic opportunities to treat the problem is extremely problematic, affecting the immune system and behavioral responses. Detailed investigation of the mechanism(s) linking the response to HIV-1 may lead to the development of new therapies. Thus, this study aims to clarify the complex network of crosstalk that exists in the neuro-immune axis. This study adds further evidence for the network and suggests that a novel factor produced by T cells may be involved in anxiety-like behavior. This indicates that it has a strong regulating effect on the

[0160] Note the behavioral phenotype of mice in which a single abnormality is higher expression of AnxA1 in T cells. Therefore, we speculate on the existence of a novel mechanism and plausible non-specific factors. The research tool will be used to further explore the specific properties of this mediator on adaptive immune responses. It was created for 16,25,26 Retrospectively, our novel observations support the idea that AnxA This is consistent with the emerging concept that AnxA1 may modulate psychiatric disorders. A genome-wide association study search (GWAS Central at www.gwascentral.org) revealed Sixty-two studies were reported on the protein, many of which were related to psychiatric disorders. In fact, reports have linked AnxA1 gene duplication to autism. 51 or in the AnxA1 gene Single nucleotide polymorphisms in schizophrenia 52 , bipolar disorder or depression 53 There is a significant connection between Most interestingly, all of these conditions are associated with immune dysfunction or inflammatory conditions. It is often related 54~56 .

[0161] Furthermore, AnxA1 is a ligand for the formyl peptide receptor (FPR) 57~60 These prototype sensors of the innate immune system were initially released by bacterial pathogens. Identified as a cellular antenna for the capture of formylated peptides 61~64 Interesting Notably, these receptors have more than one way of helping the host detect danger. Studies have shown that their expression in the olfactory system allows mice to detect infection-related This allows the animal to "sense" the presence of an olfactory stimulus and move away from the source of infection. 65~67 Other evidence also supports a role for FPR in regulating behavior. FPR1 and FP Studies in mice with knockouts of both the R2 and ALX receptors have shown that anxiety is significantly reduced. A decrease 68,69 , e.g., AnxA1 tg The opposite phenotype in mice is shown. FPR influences host behavior at both the cellular and somatic levels, and influences the external environment. This could be a prototype for a signaling molecule whose ultimate goal is to protect the host from problems. 70 .

[0162] Adoptive transfer experiments revealed a delay in recipient mice showing a significant increase in anxiety-like behavior. A similar delayed response was observed in recombinant Imood. In this case, the effect is not a direct effect on the effects of neurotransmitters in the brain, but rather the expression of genes related to anxiety. Downstream regulation of β-glucan was suggested.

[0163] AnxA1 tg Time course of occurrence of the modulating effect on anxiety by cells or after administration of Imood The imlag effect is a mechanism by which classical drugs for the treatment of depression and anxiety are shown to have clinical efficacy. This is in line with the concept that the onset of the disease is delayed by 5 to 7 days. 71 In recent research In some cases, delayed onset may allow the immune system to respond and / or adapt to the administration of these drugs. It has been suggested that this may be related to the time required for 72 , the immune system regulates gene expression This strengthens the hypothesis that cognitive control regulates emotional states and directly influences emotions.

[0164] As a novel fine-tuning factor of psychiatric disorders, Imood may have novel prognostic and diagnostic value. This will serve as a biomarker for the precise psychiatric disorder (e.g., one related to immune components) and allow patients to be identified as having the disorder. It may be possible to stratify or identify subgroups of patients who are suitable for particular drug treatments. In addition, for patients with higher Imood expression, immunomodulatory agents and Immunoglobulins such as 1B10 are recommended. Combinatorial therapy using OOD-neutralizing antibodies to achieve "healthy mind, healthy body" Thus, it may provide a unique opportunity to investigate the role of protein mediators of emotional behavior. The identification of these proteins and the availability of biological therapies that modulate their levels could be a promising approach to treating psychiatric disorders. Indeed, biologics for the treatment of psychiatric disorders target the immune system rather than the CNS. It acts specifically at the system level, preventing side effects associated with daily administration of standard treatments for psychiatric disorders. Avoiding some of the effects 73~75 .

[0165] References 1. Khandaker GM, Dantzer R, Jones PB. Psychol Med 2017; 47(13): 2229-2237. 2. Coutinho E, Vincent A. Handb Clin Neurol 2016; 133: 269-282. 3. Feinstein A, Pavisian B. Mult Scler 2017; 23(7): 923-927. 4. Pompili M et al.. J Psychosom Res 2012; 73(6): 411-417. 5. Butt AA, Evans R, Skanderson M, Shakil AO. J Hepatol 2006; 44(5): 864-868 . 6. Cournos F, McKinnon K, Sullivan G. J Clin Psychiatry 2005; 66 Suppl 6: 27 -33. 7. Bosche K, et al.. Neuropharmacology 2015; 96(Pt A): 83-93. 8. Goeb JL, Even C, Nicolas G, Gohier B, Dubas F, Garre JB. Eur Psychiatry 2 006;21(3):186-193. 9. Marin IA, Kipnis J. Neuropsychopharmacology 2017; 42(1): 28-35. 10. Dantzer R. Physiol Rev 2018; 98(1): 477-504. 11. Sheikh MH, Solito E. Annexin A1: Int J Mol Sci 2018; 19(4). 12. Leoni G, Nusrat A Biol Chem 2016; 397(10): 971-979. 13. Qin C, Yang YH, May L, Gao X, Stewart AG, Tu Y et al. Pharmacol Ther 201 5; 148: 47-65. 14. Perretti M, D'Acquisto F. Nat Rev Immunol 2009; 9(1): 62-70. 15. Bruschi M, Petretto A, Vaglio A, Santucci L, Candiano G, Ghiggeri GM. Int J Mol Sci 2018; 19(5). 16. D'Acquisto F. Curr Opin Pharmacol 2009; 9(4): 521-528. 17. Moraes LA, Ampomah PB, Lim LHK. Cell Adh Migr 2018: 1-7. 18. Boudhraa Z, Bouchon B, Viallard C, D'Incan M, Degoul F. Clin Sci (Lond) 2 016; 130(4): 205-220. 19. Biaoxue R, Xiguang C, Shuanying Y. Int J Biol Markers 2014; 29(1): e8-20. 20. Guo C, Liu S, Sun MZ. Future Oncol 2013; 9(11): 1773-1793. 21. D'Acquisto F, Piras G, Rattazzi L. Biochem Pharmacol 2013; 85(9): 1213-12 18. 22. Paschalidis N, et al. R. Cell Cycle 2010; 9(4): 784-793. 23. Paschalidis N.J Neuroinflammation 2009; 6:33. 24. Huggins A et al FASEB J 2009; 23(4): 985-996. 25. D'Acquisto F, et al Eur J Immunol 2007; 37(11): 3131-3142. 26. D’Acquisto F et al Blood 2007; 109(3): 1095-1102. 27. Huang P, Zhou Y, Liu Z, Zhang P. Mediators Inflamm 2016; 2016: 1701059. 28. Mihaylova N, et al. Autoimmunity 2017; 50(4): 257-268. 29. Tzelepis F et al J Clin Invest 2015; 125(2): 752-768. 30. Sena AA et al Sci Rep 2016; 6:31157. 31. Odobasic D et al Int Immunopharmacol 2018; 61:140-149. 32. Yazid S et al J Autoimmun 2015; 58: 1-11. 33. Yang YH et al J Immunol 2013; 190(3): 997-1007. 34. Carter DB, Kennett MJ, Franklin CL. Comp Med 2002; 52(5): 452-455. 35. Reeves WH et al Trends Immunol 2009; 30(9): 455-464. 36. Rossi B, Front Immunol 2016; 7: 506. 37. DuPage M, Bluestone JA. Nat Rev Immunol 2016; 16(3): 149-163. 38. Hirota K et al Nat Immunol 2011; 12(3): 255-263. 39. Taylor GT et al mice. Acta Neurobiol Exp (Wars) 2017; 77(3): 254-260. 40. Bourin M, Hascoet M. Eur J Pharmacol 2003; 463(1-3): 55-65. 41. Deacon RM, Rawlins JN. Behav Brain Res 2005; 156(2): 241-249. 42. Brooks SP, Dunnett SB. Nat Rev Neurosci 2009; 10(7): 519-529. 43. Filiano AJ, et al Nature 2016; 535(7612): 425-429. 44. Brachman RA, et al J Neurosci 2015; 35(4): 1530-1538. 45. Rattazzi L et al Transl Psychiatry 2013; 3: e280. 46. Hovatta I et al. Nature 2005; 438(7068): 662-666. 47. Engin E et al Pharmacol Ther 2012; 136(2): 142-152. 48. Bates MK et al Biotechniques 2006; 40(2): 199-208. 49. Miller AH, Haroon E, Felger JC Neuropsychopharmacology 2017; 42(1): 334-3 59. 50. Haroon E, Raison CL, Miller AH. Neuropsychopharmacology 2012; 37(1): 137- 162. 51. Correia CT et al Mol Autism 2014; 5(1): 28. 52. Clark D et al Mol Psychiatry 2006; 11(5): 459-470, 423. 53. Huang J et al. Am J Psychiatry 2010; 167(10): 1254-1263. 54. Severance EG et al Pharmacol Ther 2018; 189:184-198. 55. Prata J et al J Neuroinflammation 2017; 14(1): 179. 56. Muller N. Dialogues Clin Neurosci 2017; 19(1): 55-63. 57. Cooray SN et al Proc Natl Acad Sci USA 2013; 110(45): 18232-18237. 58. Bena S et al J Biol Chem 2012; 287(29): 24690-24697. 59. Pederzoli-Ribeil M et al Blood 2010; 116(20): 4288-4296. 60. Gastardelo TS et al. Am J Pathol 2009; 174(1): 177-183. 61. Winther M et al Basic Clin Pharmacol Toxicol 2018; 122(2): 191-198. 62. Chen K et al J Autoimmun 2017; 85:64-77. 63. Lee HY et al J Cell Biochem 2017; 118(6): 1300-1307. 64. Migeotte I et al Cytokine Growth Factor Rev 2006; 17(6): 501-519. 65. Bufe B et al J Biol Chem 2012; 287(40): 33644-33655. 66. Ackels T et al. Front Neuroanat 2014; 8:134. 67. Stempel H et al J Biol Chem 2016; 291(18): 9762-9775. 68. Gao JL et al. Behav Genet 2011; 41(5): 724-733. 69. Gallo I et al. PLoS One 2014; 9(12): e114626. 70. Dietschi Q et al Proc Natl Acad Sci USA 2017; 114(28): 7397-7402. 71. Lam RW. Eur Neuropsychopharmacol 2012; 22 Suppl 3: S492-498. 72. Kiecolt-Glaser JK et al Annu Rev Psychol 2002; 53:83-107. 73. Serretti A, Chiesa A. Clin Pharmacol Ther 2011; 89(1): 142-147. 74. Uzun S et al Psychiatr Danub 2010; 22(1): 90-93. 75. Babic D et al Psychiatr Danub 2010; 22(1): 75-78.

[0166] array 001 mwklgrgrvl ldeppeeedg lrggpppaaa aaaqaqvqga sfrgwkevts lfnkddeqhl 061 lerckspksk gtnlrlkeel kaekksgfwd nlvlkqniqs kkpdeiegwe ppklaledis 121 adpedtvggh pswsgwedda kgstkytsla ssanssrwsl raagrlikev winfsqliis 181 frkhclahyr elrlciky SEQ ID NO: 1 001 mwklgrgrvl ldeppeeedg lrggpppaaa aaaqaqvqga sfrgwkevts lfnkddeqhl 061 lerckspksk gtnlrlkeel kaekksgfwd nlvlkqniqs kkpdeiegwe ppklaledis 121 adpedtvggh pswsgwedda kgstkytsla ssanssrwsl raagrlvsir rqskghltds 181 peeae SEQ ID NO: 2 NVAMY SEQ ID NO: 3 (1B10 VHCDR1) RIRSKANNYATYYADSVKG SEQ ID NO: 4 (1B10 VHCDR2) WVIVPLYFDY SEQ ID NO: 5 (1B10 VHCDR3) RSSKSLLSSKGITSLY SEQ ID NO: 6 (1B10 VLCDR1) KSLLSSKGITS SEQ ID NO: 7 (1B10 VLCDR1) XMSNLAS (wherein X is independently any amino acid) SEQ ID NO: 8 (1B10 VLCDR2) QMS SEQ ID NO: 9 (1B10 VLCDR2) GHRLQTPFT SEQ ID NO: 10 (1B10 VLCDR3) AVQLVESGGGLVQPEESLKISCAASGITFS NVAMY WVRQAPGKGLEWVA RIRSKANNYATYYADSVKG RFTISRDDSKSM VYLQMDNLKTEDTAMYYCSA WVIVPLYFDY WGQGVMVTVSS SEQ ID NO: 11 (1B10 VH) 001 gcggtgcagc tggttgagtc tggtggagga ttggtgcagc ctgaggagtc attgaaaatc 061 tcatgtgcag cttctggaat caccttcagt aatgttgcca tgtactgggt ccgccaggct 121 ccaggaaagg gtctggaatg ggttgctcgc ataagaagta aagctaataa ttatgcaaca 181 tattatgctg attcagtgaa aggcagattc accatctcca gagatgattc aaaaagcatg 241 gtctacctac aaatggataa cttgaaaact gaggacacag ccatgtacta ttgttcagca 301 tgggttatag tgcccctata ttttgattac tggggccaag gagtcatggt cacagtctcc 361 tca SEQ ID NO: 12 (1B10 VH coding) DIVMTQAPLSVSVTPGESASISC RSSKSLLSSKGITSLY WYLQRPGKSPQLLIYX1 MSNLAS GVPDRFSX2SGSETDFTL KISX3VEAEDVGVYYC GHRLQTPFT FGSGTKLEIK (wherein X1 to X3 are independently any amino acid, X1 is Q, X2 is S, and X3 is K) (preferably SEQ ID NO: 13 (1B10 VL) DIVMTQAPLSVSVTPGESASISCRSS KSLLSSKGITS LYWYLQRPGKSPQLLIY QMS NLASGVPDRFSSSGSETDFTLKI SKVEAEDVGVYYC GHRLQTPFT FGSGTKLEIK SEQ ID NO: 14 (1B10 VL) 001 gatattgtga tgactcaagc tccactctct gtatctgtca ctcctggaga gtcagcttcc 061 atctcctgca grtctagtaa gagtctgcta agtagtaagg gcatcacttc cttgtattgg 121 taccttcaga ggccaggaaa gtctcctcag ctcctgatat atcrgatgtc caaccttgcc 181 tcaggagttc cagacaggtt tagtrgcagt gggtcagaaa ccgattttac actgaaaatc 241 agtarggtgg aggctgagga tgttggtgtt tattactgtg gacatcgtct acaaactcca 301 ttcacgttcg gctcagggac gaaattggaa ataaaa (wherein r is independently any nucleotide) SEQ ID NO: 15 (1B10 VL coding) 001 gatattgtga tgactcaagc tccactctct gtatctgtca ctcctggaga gtcagcttcc 061 atctcctgca ggtctagt AA GAGTCTGCTA AGTAGTAAGG GCATCACTTC Cttgtattgg 121 taccttcaga ggccaggaaa gtctcctcag ctcctgatat at CAGATGTC C aaccttgcc 181 tcaggagttc cagacaggtt tagtagcagt gggtcagaaa ccgattttac actgaaaatc 241 agtaaggtgg aggctgagga tgttggtgtt tattactgt G GACATCGTCT ACAAACTCCA 301 TTCACG ttcg gctcagggac gaaattggaa ataaaa SEQ ID NO: 16 (1B10 VL coding; CDRs underlined) WKLSRSRVLLDEPPEEEDVLRGAPPASAAAPASVRARVGAQGASLRGWKEATSLFNKDDEEHLLETSRSPKSKGTNQRLR EELKAEKKSGFWDALVLKQNAQPKKPDQIEGWEPPKLTAEDVVADHTEDDRSGCPPWSAWEDDTKGSTKYTSLANSASSS RWSLRSAGKLVSIRRQSKGHLTETCEEGE SEQ ID NO: 17 (2610019F03Rik (aa 2-190)) GFTFSDYN SEQ ID NO: 18 (1C4 VHCDR1) IIYDGDRT SEQ ID NO: 19 (1C4 VHCDR2) ATGLAY SEQ ID NO: 20 (1C4 VHCDR3) QSLLYSENKKNY SEQ ID NO: 21 (11C4 VLCDR1) WAS SEQ ID NO: 22 (1C4 VLCDR2) QQYYNFPST SEQ ID NO: 23 (1C4 VLCDR3) EVQLVESGGGLVQPGRSLKLSCAAS GFTFSDYNMAWVRQAPKRGLEWVAT IIYDGDRT YYRDSVKGRFTISRDKAKTTLY LQMDSLRSEDTATYYC ATGLAY WGQGTLVTVSS SEQ ID NO: 24 (1C4 VH; CDRs underlined) 001 atggacatca ggctcagctt ggttttcctt gtccttttca taaaaggtgt ccagtgtgag 061 gtgcagctgg tggagtctgg cggaggctta gtacagcctg gaaggtccct gaaactctcc 121 tgtgcagcct ca ggattcac tttcagtgac tataac atgg cctgggtccg ccaggctcca 181 aagaggggtc tggagtgggt cgcaacc att atttatgatg gtgataggac t tactatcga 241 gactccgtga agggccgatt cactatctcc agagataaag caaaaaccac cctatatttg 301 caaatggaca gtctgaggtc tgaggacacg gccacttatt actgt gcaac agggcttgct 361 tac tggggcc aaggcactct ggtcactgtc tcttcag SEQ ID NO: 25 (1C4 VH coding; CDRs underlined) DIVMTQTPSSQAVSPGEKVTMSCKSS QSLLYSENKKNY LAWYQQKPGQSPKLLIY WAS TRESGVPDRFIGSGSGTDFTLT ISSVQAEDLAVYYC QQYYNFPST FGTGTTLELK SEQ ID NO: 26 (1C4 VL CDRs underlined) 001 atggaatcac agacccaggt cctcatgtcc ctgctgctct gggtatctgg tacctgtggg 061 gacattgtga tgacccagac tccatcctcc caggctgtgt caccagggga gaaggtcact 121 atgagctgca agtccagt ca gagtctttta tacagtgaaa acaaaaagaa ctac ttggcc 181 tggtaccagc agaaaccagg gcagtctcct aaactgctga tctac tgggc atcc actagg 241 gaatctgggg tccctgatcg cttcataggc agtggatctg ggacagattt cactctgacc 301 atcagcagtg tgcaggcaga agacctggct gtttattact gc cagcaata ctataacttt 361 ccgagcacg t ttggaactgg gaccacgctg gagctgaaac Sequence number 27 (underlined in the 1C4 VL coding CDR)

Claims

1. A method for treating a psychiatric disorder, comprising administering a testicular development-related protein (TDRP) antagonist The method comprises administering to an individual in need thereof a compound comprising administering to said individual ...

2. 10. The method of claim 1, wherein the psychiatric disorder is characterized by anxiety-related behavior.

3. 3. The method of claim 1 or 2, wherein the psychiatric disorder is an anxiety disorder.

4. The method according to any one of claims 1 to 3, wherein the psychiatric disorder is obsessive-compulsive disorder (OCD). Law.

5. The method of any one of claims 1 to 4, wherein the individual has an immunopathology.

6. The method of claim 5, wherein the individual is undergoing treatment for an immune condition.

7. 7. The method of claim 5 or 6, wherein the immunopathology is an autoimmune disorder.

8. 8. The method of claim 5, wherein the individual is undergoing treatment with an immunosuppressant. method.

9. The TDRP antagonist reduces the expression, level, or amount of TDRP in the circulatory system of the individual. The method according to any one of claims 1 to 8, wherein the concentration of the compound is reduced.

10. 10. The method of claim 1, wherein the TDRP antagonist is an antibody molecule that specifically binds to TDRP.

10. The method according to any one of claims 1 to 9.

11. The method of any one of claims 1 to 10, wherein the antibody molecule is 1B10 or 1C4. Law.

12. An antibody molecule that specifically binds to TDRP, comprising a VH domain and a VL domain. an antigen-binding site, wherein the VH domain is a VHCDR1 of SEQ ID NO: 3 or a variant thereof, VHCDR2 of SEQ ID NO: 4 or a variant thereof, and VHCDR3 of SEQ ID NO: 5 or a variant thereof including antibody molecules.

13. The VL domain is a VLCDR1 of SEQ ID NO: 6 or a variant thereof, a VL CDR2 or a variant thereof, and VLCDR3 or a variant thereof of SEQ ID NO:

10. Muka, or The VL domain is a VLCDR1 of SEQ ID NO: 7 or a variant thereof, a VL CDR2 or a variant thereof, and VLCDR3 or a variant thereof of SEQ ID NO:

10. nothing, The antibody described in claim 12.

14. 12 or 13, wherein the VH domain comprises the amino acid sequence of SEQ ID NO: 11 or a variant thereof.

14. The antibody molecule according to 13.

15. The VL domain has the amino acid sequence of SEQ ID NO: 13 or SEQ ID NO: 14, or a variant thereof The antibody molecule of any one of claims 12 to 14, comprising a nucleotide sequence.

16. An antibody molecule that specifically binds to TDRP, comprising a VH domain and a VL domain. an antigen-binding site, wherein the VH domain is VHCDR1 of SEQ ID NO: 18 or a variant thereof; VHCDR2 or a variant thereof of sequence number 19, and VHCDR3 or a variant thereof of sequence number 20 and the VL domain comprises a VLCDR1 of SEQ ID NO: 21 or a variant thereof, a VLCDR1 of SEQ ID NO: 2 2 or a variant thereof, and VLCDR3 of SEQ ID NO: 23 or a variant thereof. Hmm, antibody molecules.

17. 17. The method of claim 16, wherein the VH domain comprises the amino acid sequence of SEQ ID NO: 24 or a variant thereof. The antibody molecule described.

18. 16 or 17, wherein the VL domain comprises the amino acid sequence of SEQ ID NO: 26 or a variant thereof.

18. The antibody molecule according to 17.

19. The TDRP antagonist is an antibody molecule according to any one of claims 12 to 18. The method according to any one of claims 1 to 11,

20. TDRP antagonists for use in methods of treating psychiatric disorders.

21. The use according to claim 20, wherein the method is a method according to any one of claims 1 to 11. TDRP antagonists for use.

22. The TDRP antagonist is an antibody molecule according to any one of claims 12 to 18.

22. The TDRP antagonist for use according to claim 20 or 21,

23. Use of TDRP antagonists in the manufacture of medicaments for use in methods of treating psychiatric disorders For.

24. The use according to claim 23, wherein the method is a method according to any one of claims 1 to 11. For.

25. The TDRP antagonist is an antibody molecule according to any one of claims 12 to 18.

25. The use according to claim 23 or 24.

26. Pharmaceutical compositions comprising a therapeutically effective amount of a TDRP antagonist and a pharmaceutically acceptable excipient thing.

27. The TDRP antagonist is an antibody molecule according to any one of claims 12 to 18. The pharmaceutical composition of claim 26.

28. The method according to claim 26 or 27, which is used in the method according to any one of claims 1 to 11. A pharmaceutical composition comprising:

29. Diagnosing a mental disorder in an individual or identifying individuals at high risk of suffering from a mental disorder; Or a method for assessing anxiety in an individual, comprising: Determining the expression, level, amount or concentration of TDRP in a sample obtained from an individual. Including, a method.

30. 1. A method of monitoring an individual undergoing treatment, comprising: Expression, level, amount or concentration of TDRP in a sample obtained from an individual receiving treatment determining a

31. 31. The method of claim 30, wherein the individual is undergoing treatment for an immune condition.

32. 31. The method of claim 30, wherein the individual is undergoing treatment for a psychiatric disorder.

33. 33. Any one of claims 30 to 32, wherein the sample is a blood, serum or plasma sample. The method described below.

34. The method of any one of claims 30 to 33, wherein the sample comprises PBMCs.

35. determining the expression of TDRP in a sample of PBMCs obtained from said individual. The method of claim 34.

36. A method for screening a compound having therapeutic activity against a psychiatric disorder, comprising: determining binding of the test compound to the isolated TDRP; Binding to TDRP is an indication that the compound has therapeutic activity for treating a psychiatric disorder. How to do it.

37. A method for screening a compound having therapeutic activity against a psychiatric disorder, comprising: Effect of test compounds on the expression, level, amount or concentration of TDRP in non-human mammals determining the impact of the A decrease in the expression, level, amount or concentration of TDRP indicates that the compound has therapeutic activity against psychiatric disorders. A method that serves as an indicator of sexuality.

38. 38. The method of claim 36 or 37, wherein the activity is anxiolytic activity.

39. determining the effect of said test compound on anxiety-related behavior in a non-human mammal; The method of any one of claims 36 to 38, further comprising:

40. 1. A method for determining T cell activation in an individual, comprising: determining the expression of TDRP in a sample of T cells obtained from said individual. ,method.

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