POFUT1 gene determines tissue-infiltrating CD4+ T cell function and related biomarkers in disease

Abstract

This invention discloses the role of the POFUT1 gene in determining the function of tissue-infiltrating CD4+ T cells and the application of related biomarkers in diseases, belonging to the field of biomedical technology. This invention discovers that POFUT1 is a decisive gene regulating the development of CXCR5+CD4 T cells; simultaneously, CXCR5+ and SLAMF1+CD4 T cells, as tissue-infiltrating source cells, possess the function of carrying out CD4 T cell-mediated antigen-specific killing and tissue destruction, leading to the pathogenesis of autoimmune diseases. Related applications include: using the POFUT1 gene, CXCR5+, and SLAMF1+CD4 T cells as biomarkers for the clinical diagnosis and treatment of diseases caused by tissue-infiltrating CD4 T cells, such as autoimmune diseases and xenotransplant rejection. The killing function of CXCR5+ and SLAMF1+CD4 T cells as tissue-infiltrating source cells can also be used to treat tumors.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, specifically the role of the POFUT1 gene in determining the function of tissue-infiltrating CD4+ T cells and the application of related biomarkers in diseases. Background Technology

[0002] In recent years, immunotherapy (including cell therapy) has flourished. Research and applications have largely focused on using immune cells with direct killing functions to kill cancer cells, such as CD8 T cells, NK cells, and promoting antibody production by B cells. Similarly, in autoimmune research, autoantibodies that recognize self-antigens have long been considered key to disease. Therefore, autoantibodies and B cells that produce them have attracted significant attention. As the command center of the adaptive immune system, CD4 T cells are generally believed to function by activating and regulating CD8 T cells and assisting B cells in antibody production. Therefore, the preferred treatment or application has focused on CD8 T cells, B cells, or antibody-mediated NK cells. However, CD4 T cells can function even without these cells. Therefore, targeting CD4 T cells deserves greater attention, whether for using immune cells to kill tumors or suppressing immune cells to treat autoimmune diseases.

[0003] T cells constitute a complex population. Antigen-specific T cells comprise only one in a million of the total. Within this one in a million, there are regulatory T cells that suppress the immune response, and exhausted T cells that have lost their function. Truly functional effector cells are extremely rare. Current cell therapies targeting cancer cells activate all T cells. Therefore, this involves proportionally expanding all T cells without selectively increasing the desired cells. Based on current cell therapy manufacturing processes, CD8 T cells are easily activated and become the main proliferating cells. While antigen-specific CD8 T cells do possess killing capabilities, their function is singular, easily suppressed by CD4 regulatory cells with the same antigen specificity, and can rapidly die from overactivation, becoming exhausted T cells. CD4 T cells, on the other hand, act like commanders, employing diverse mechanisms, achieving killing through CD8 or by activating innate immune cells such as macrophages. Therefore, CD4 T cell-mediated antigen-specific killing and tissue destruction involves multiple immune cells, similar to a multi-armed force operating in coordinated warfare. Existing CAR-T therapies, both in vivo and in vitro, are limited by antigens, failing to fully utilize the multi-antigen specificity of T cells; they also lack the crude process of simply proliferating all T cells; and their tissue infiltration capacity is very limited. Therefore, for solid tumors, the most effective option is to find antigen-specific CD4 effector T cells with tissue infiltration capacity and use them for treatment.

[0004] CXCR5 is a chemokine receptor that has long been a target for cancer metastasis research. In recent years, CXCR5-positive CD4 T cells have been defined as follicular B helper T cells (TFH). Our understanding of them is primarily focused on helping B cells express antibodies. Especially in autoimmune diseases and transplant immunology research, it is believed that clearing B cells and antibodies can eliminate the disease. However, the function of CXCR5+CD4 cells themselves, without the assistance of B cells, is not clearly understood. Most notably, in autoimmune diseases and transplant immunology, the understanding of T cells is largely limited to helping B cells produce pathogenic antibodies. Little is known about the role of CD4 T cells themselves, without the assistance of B cells.

[0005] Signaling lymphocyte activation molecule family member 1 (SLAMF1) is expressed on the surface of various cells and has long served as a cancer biomarker. However, the function of SLAMF1+CD4 cells in autoimmune diseases and transplant immunity remains unclear.

[0006] Experimental autoimmune encephalomyelitis (EAE) is a widely used animal model for evaluating therapeutics against autoimmune-mediated central nervous system (CNS) diseases. Myelin oligodendrocyte glycoprotein (MOG) is a member of the immunoglobulin superfamily and possesses high immunogenicity. Located in the outermost layer of myelin in the CNS, MOG is a potential target for the binding of MOG antibodies and specific immune cells. In mice with MOG-induced EAE, severe inflammation of the central nervous system occurs, leading to the killing of myelin cells and tissue destruction. This results in damage to the myelin sheath structure, forming typical demyelinating pathological changes. The MOG35-55 peptide-induced EAE model is actually a pathogenic model that does not require the participation of B cells or antibodies. Therefore, EAE is also a classic model for specific CD4 T cell research. By evaluating the performance of MOG-specific CD4 T cells, this study elucidates the pathological aspects, particularly regarding autoimmune diseases, inflammation, and the killing and tissue destruction functions mediated by antigen-specific CD4 T cells, thereby inferring the mechanisms of action and applications of other antigen-specific CD4 T cells.

[0007] Protein O-Fucosyltransferase 1 (EC:2.4.1.221; POFUT1) is a glycosyltransferase that provides post-glycosylation modification after protein synthesis. POFUT1 glycosyltransferase has long been recognized as a key component of the NOTCH cellular signaling pathway and has been targeted for tumor cell research.

[0008] However, it is unknown whether POFUT1 can affect the function of CD4+ T cells, and the mechanism by which it affects autoimmune diseases through CD4+ T cells is unclear. Summary of the Invention

[0009] In view of the above-mentioned prior art, the present invention provides a method for the POFUT1 gene to determine the function of tissue-infiltrating CD4+ T cells and the application of related biomarkers in diseases. The purpose is to reveal the function of POFUT1 glycosyltransferase in CD4 T immune cells and its mechanism of action in autoimmune diseases.

[0010] The first technical solution provided by this invention is the application of protein O-fucosyltransferase POFUT1 or its encoded gene as a target in the preparation of drugs for the prevention, relief and / or treatment of autoimmune diseases.

[0011] In some embodiments, the application is to prevent, alleviate, and / or treat autoimmune diseases by inhibiting the expression of the protein O-fucosyltransferase POFUT1 or its encoded gene in CD4 T cells.

[0012] In some embodiments, the autoimmune diseases include, but are not limited to, systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, ulcerative colitis, and xenotransplant rejection.

[0013] In some embodiments, the drug includes an inhibitor that suppresses the expression of the protein O-fucosyltransferase POFUT1 in CD4 T cells.

[0014] In some embodiments, the inhibitor includes, but is not limited to, interfering RNA, gene knockout agents, or chemical inhibitors. Preferably, the inhibitor may be siRNA.

[0015] In some implementations, the application includes at least one of the following functions:

[0016] (1) Inhibits the infiltration ability of SLAMF1-positive (+) CD4 T cells; (2) Block the development of CXCR5-positive (+) CD4 T cells.

[0017] In some embodiments, the dosage form of the drug can be a medically acceptable conventional dosage form. As a preferred example, without limitation, it can be an injection, a lyophilized powder for injection, a suspension, an implant, an embolization, a capsule, a tablet, a pill, or an oral liquid.

[0018] In some embodiments, the drug contains pharmaceutical excipients.

[0019] In some embodiments, the pharmaceutical excipients include at least one of solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, and wetting agents.

[0020] In some embodiments, the pharmaceutical excipients further comprise at least one of the following: osmotic pressure regulators, stabilizers, flow aids, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesion agents, integrators, penetration enhancers, pH adjusters, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, encapsulating agents, humectants, flocculants and anti-flocculators, filter aids, or release inhibitors.

[0021] In some embodiments, the drug contains a pharmaceutical carrier.

[0022] In some embodiments, the pharmaceutical carrier is selected from microcapsules, microspheres, nanoparticles, or liposomes.

[0023] The second technical solution provided by this invention is the application of protein O-fucosyltransferase POFUT1 or its encoded gene as a target in the preparation of an inhibitor for autoimmune diseases.

[0024] In some embodiments, the blocking agent can bind to the target site to inhibit the expression of the protein O-fucosyltransferase POFUT1 in CD4 T cells.

[0025] In some embodiments, the autoimmune diseases include, but are not limited to, systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, ulcerative colitis, and xenotransplant rejection.

[0026] The third technical solution provided by this invention is the application of an inhibitor of protein O-fucosyltransferase POFUT1 in the preparation of a drug for treating diseases caused by tissue-infiltrating CD4 T cells.

[0027] In some embodiments, the application involves inhibiting the expression of POFUT1 or its encoded gene within CD4 T cells, blocking the development of CXCR5-positive CD4 T cells, and inhibiting the infiltration ability of SLAMF1-positive CD4 T cells. This invention provides a fourth technical solution: the application of CXCR5-positive CD4 T cells and / or SLAMF1-positive CD4 T cells as targets in the preparation of drugs for the prevention, alleviation, and / or treatment of autoimmune diseases.

[0028] In some embodiments, the drug prevents, alleviates, and / or treats autoimmune diseases by blocking the development of CXCR5-positive CD4 T cells and inhibiting the infiltration of SLAMF1-positive CD4 T cells.

[0029] In some embodiments, the autoimmune diseases include, but are not limited to, systemic lupus erythematosus, lupus nephritis, rheumatoid arthritis, multiple sclerosis, ulcerative colitis, and xenograft rejection.

[0030] Compared with the prior art, the beneficial effects of the present invention are as follows: This invention discovers that POFUT1 is a key gene regulating the development of CXCR5-positive CD4 T cells; it also reveals that CXCR5+ and SLAMF1+ CD4 T cells, as tissue-infiltrating source cells, determine the pathogenesis of autoimmune diseases caused by antigen-specific CD4 T cells. Applications arising from these findings include: POFUT1 gene, CXCR5-positive, and SLAMF1+ CD4 T cells, as biomarkers, can be used in the clinical diagnosis and treatment of diseases caused by tissue-infiltrating CD4 T cells, including autoimmune diseases such as multiple sclerosis, inflammatory bowel disease-ulcerative colitis, lupus nephritis, rheumatoid arthritis, and xenograft rejection. Furthermore, CXCR5-positive and SLAMF1+ CD4 T cells extracted from secondary lymphoid tissue can serve as a source of tissue-infiltrating immune cells for invading tissues with specific antigenic environments, including tumors. Attached Figure Description

[0031] Figure 1 The curves show the survival of mice after EAE syndrome was induced, with ≥ 6 mice in each group.

[0032] Figure 2 This is the survival curve of mice after EAE disease was induced. Conditional knockout of the POFUT1 gene in CD4 T cells prevented mice from developing EAE disease, with ≥10 mice in each group.

[0033] Figure 3 Wild-type mice and CD4crePOFUT1 conditional knockout mice were immunized with MOG35-55. Seven days later, draining lymph node tissue cells were collected for staining analysis, with ≥ 6 cells per group.

[0034] Figure 4 Conditional knockout of the POFUT1 gene in CD4 T cells impairs the differentiation and development of CXCR5-positive and Bcl6-positive Tfh cells in mice; CD4 T cells with POFUT1 gene knockout have normal proliferative capacity and the ability to express cytokines, with ≥ 6 cells per group of mice.

[0035] Figure 5 After immunizing CD90.1 wild mice with MOG 35-55, the CXCR5-positive CD4 T cells in these mice were then transfused into non-CD90.1 CD4crePOFUT1 mice. The transfused mice were successfully induced with EAE symptoms, with ≥ 6 mice per group.

[0036] Figures 6-8 The analysis of central nervous system cells after mouse sacrifice showed that donor cells accounted for 90% of the infiltrated CD4 T cells in the central nervous system of diseased mice, with ≥6 in each group of mice.

[0037] Figure 9 Wild-type mice and CD4crePOFUT1 conditional knockout mice were immunized with MOG35-55. Seven days later, draining lymph node tissue cells were collected for staining analysis, with ≥ 4 cells per group.

[0038] Figure 10 The CD4crePOFUT1 conditional knockout mice showed the presence of SLAM+CD4 cells, with a slight increase; conversely, the CXCR5 cell population was absent, with ≥ 4 cells per group of mice.

[0039] Figure 11 The method involved immunizing CD90.1 wild mice with MOG 35-55, then transferring the SLAM+ positive CD4 T cells from these mice into non-CD90.1 CD4crePOFUT1 mice. The transfused mice were successfully induced with EAE symptoms, with ≥6 mice per group.

[0040] Figures 12-14 The analysis of central nervous system tissue cells after mouse sacrifice revealed that, in the central nervous system of diseased mice, only 10% of the infiltrated CD4 T cells were transfused donor cells. The remaining 90% consisted of drained donor CD4 T cells, with ≥ 6 cells per group of mice. Detailed Implementation

[0041] refer to Figures 1-14 The preferred embodiments of the present invention will be described below. It should be understood that the embodiments are for better explanation of the present invention and are not intended to limit the present invention. All data listed in the present invention have been successfully replicated more than twice in biological experiments.

[0042] Test method: 1. EAE mouse model scoring system: Approximately one week after immunization, mice developed clinical symptoms. Daily observations were conducted, and mice were scored using the following scoring system: EAE model scoring criteria: 0 points, normal; 1 point, drooping tail; 2 points, weakness in one hind limb, unsteady gait, recoverable after passive rolling; 3 points, paralysis or near-paralysis of both hind limbs, irreversible after passive rolling, but able to move upon stimulation; 4 points, paralysis and dragging of both hind limbs, no response to stimulation, muscle weakness accompanied by urinary and fecal incontinence; 5 points, quadriplegia / death. The mouse symptom scores were plotted against time. A clinical score of 3 or higher was considered severe.

[0043] 2. Mouse tissue cell isolation, staining, and flow cytometry analysis: Mouse spleen, draining lymph nodes, and central nervous system tissue (brain and spine) were homogenized by crushing. After passing the homogenates through a 70-micron sieve, spleen and lymph node cell suspensions were prepared. The central nervous system tissue homogenate was centrifuged at a 70:30 Percoll gradient, and the precipitate was used to prepare cell suspensions. All cell suspensions were lysed with erythrocytes for staining.

[0044] The flow cytometers used for analysis were BD LSR II and Fortessa. The cell sorting instrument was BD Aria II.

[0045] Raw materials used in the examples: All mice used in this patent experiment were on a B6 background. Mouse sources: POFUT1 lox / lox mice were custom-made. Wild-type mice C57 / B1.6 (B6) were purchased from Taconic. B-cell knockout mice (CD19KO), CD4-Cre, and CD90.1 transgenic mice were purchased from Jackson Laboratories. Using these mice, all mouse species used in this patent experiment were obtained through hybridization, including CD4crePOFUT1 mice on both CD90.1 and non-CD90.1 backgrounds.

[0046] EAE Immunosuppressant: Complete Freund's Adjuvant (Sigma), MOG35-55 (30 ug), and 1 mg mycobacteria (Difco), 200 ng pertussis toxin (Toxin Technologies). The antibody clone information used for cell staining is as follows: Antibodies from Biolegend: CD45 (30-F11), CD3ε (145-2C11), CD4 (GK1.5), CD11b (M1 / 70), CD45.1 (A20), CD90.1 (OX-7), TCRVβ11 (KT11), IFN-γ (XMG1.2), IL-17a (TC11-18H10.1). Antibodies from BD: CXCR5 (2G8), Bcl-6 (K112-91), GM-CSF (MP1-22E9). Cell viability staining was performed using the Zombie Aqua™ Fixed Viability Kit (Biolegend). Before intracellular cytokine staining, cells were activated with PMA and Ionomycin (Sigma) and incubated with Golgi-stop (BD).

[0047] Cell enrichment magnetic beads were sourced from Miltenyi Biotec, BD, and Biolegend. Example 1 The experimental autoimmune encephalomyelitis (EAE) model in mice, induced by the MOG 35-55 oligodendrocyte glycoprotein peptide (MOG 35-55) as a specific antigen, is a classic animal model simulating multiple sclerosis. The pathogenic mechanism of the MOG 35-55-induced EAE model involves the induction of MOG 35-55 antigen-specific immunity, primarily through CD4 effector T cells invading the central nervous system (CNS), leading to inflammation, demyelination, and neurodegenerative changes, accompanied by early disruption of the blood-brain barrier. The involvement of B cells and the action of autoantibodies are not required. Following antigen immunization, MOG 35-55-specific CD4 T cells are activated and differentiate in secondary lymphoid tissues; after maturing into effector T cells, they invade the CNS, becoming tissue-infiltrating CD4 T cells. Therefore, this model has also been used to study the development, differentiation, and pathogenic function of CD4 effector T cells.

[0048] A total of 30 μg of complete Freund's adjuvant was mixed with 30 μg of MOG 35-55 mice and administered bilaterally to the abdomen of adult C57BL / 6 (B6) background wild-type (WT) mice, B-cell knockout mice (CD19KO), and CD4crePOFUT1 conditional knockout mice (obtained through a cross between CD4Cre mice and POFUT1 FLOX / FLOX mice). Results are as follows: Figures 1-2 As shown, since the MOG 35-55-induced EAE pathogenic model in mice does not require the participation of B cells, similar to wild-type mice, B cell knockout mice can also be successfully induced. Therefore, in this MOG 35-55-induced EAE pathogenic model, the killing of target cells expressing MOG and tissue destruction is not actually antibody-mediated, but rather CD4 T cell-mediated. Conditional knockout of the POFUT1 gene within CD4 T cells prevents the induction of EAE symptoms in mice. Through various CD4 T cell conditional knockout mouse experiments (including standard knockout model, 2D2 knockout model, FOXP3GFP model, Treg clearance model, MOG 1-125 model, cell transfer model, and their replicates, with a cumulative total of over 100 CD4 T cells conditionally knocked out of POFUT1, only 1-2 mice had a clinical score of 0.5 or 1, and the remaining mice did not show any symptoms. Data is not fully listed). Wild-type mice all exhibited EAE symptoms, with the highest average score exceeding 3 points. Figures 3-4 In some experimental groups, the score even reached 4 points. Figures 5-8(11-14). Because the applicant's institution does not allow mice to die from the disease, mice with a disease score of 4 must be euthanized; 4 is essentially the maximum score. Compared to wild-type mice that develop severe illness or even die after EAE, mice with conditional knockout of the POFUT1 gene in CD4 T cells show less than 1% mild symptoms.

[0049] Example 2 A total of 30 μg / mouse MOG 35-55 was administered to the bilateral abdomen of adult CD90.1 wild-type mice, mixed with complete Freund's adjuvant. Seven days later, lymph nodes were drained, and a cell suspension without erythrocytes was prepared. Figures 3-4 As shown, conditional knockout of the POFUT1 gene in CD4 T cells inhibited the differentiation and development of mouse CXCR5-positive and Bcl-6-positive Tfh cells, with the proportion of this cell population decreasing by more than tenfold. However, CD4 T cells with the POFUT1 gene knocked out exhibited normal proliferative capacity and the ability to express cytokines.

[0050] A CD4 T cell negative selection magnetic bead reagent was used to remove non-CD4 T cell leukocytes. CXCR5-positive cells within the CD4 T cell population were labeled with biotin-conjugated CXCR5 antibody; then, biotin-specific streptavidin magnetic beads were used to label biotin-conjugated to the CXCR5 antibody; CXCR5-positive CD4 T cells were obtained through positive sorting. The CXCR5-positive CD4 T cells obtained using this method exhibited a relatively high proportion of MOG35-55 specific cells.

[0051] A dose of 2 million cells per mouse was transfused via blood into non-CD90.1 CD4crePOFUT1 conditional knockout mice. Twenty-four hours later, immunization was performed at the base of the tail of mice using a mixture of 30 μg / mouse MOG 35-55 and complete Freund's adjuvant and pertussis toxin. CD4crePOFUT1 conditional knockout mice without transfusion of CXCR5-positive CD4 T cells served as negative controls, while wild-type mice served as positive controls. Figures 5-8As shown, Group 1 used CD4crePOFUT1 conditional knockout mice without CXCR5-positive CD4 T cells as a negative control, with a mean symptom score of 0; Group 2 used wild-type mice as a positive control, with a maximum mean symptom score of 4; Group 3, CD4crePOFUT1 conditional knockout mice that were transfused with CXCR5-positive CD4 T cells 48 hours in advance, were successfully induced with EAE symptoms, with a maximum mean symptom score of 2. Furthermore, in the central nervous system of the affected mice, 90% of the infiltrating tissues were composed of transfused donor cells (CD90.1+ wild-type CD4 T cells). These donor cells contained over 25% expressing cytokines and inflammatory factors (IFNγ, IL7, and GM-CSF), exhibiting typical effector cell characteristics. These results indicate that CXCR5+ CD4 cells invade the mouse central nervous system from the bloodstream, becoming infiltrating cells with high proliferative capacity.

[0052] Based on the results of this embodiment, we can infer that: 1. MOG-specific CXCR5+CD4 cells can invade central nervous system tissues, inducing various types of sclerosis and similar autoimmune diseases. Similar to the MOG-specific CXCR5+CD4 cells in this embodiment, other antigen-specific CXCR5+CD4 cells can invade autologous or transplanted organ tissues, carrying out CD4 T cell-mediated killing and tissue destruction against antigen-specific target cells. For example, the kidneys in lupus nephritis; joint tissues in rheumatoid arthritis; and various organs in xenotransplantation. Therefore, they can be used as biomarkers for clinical diagnosis and treatment. 2. Utilizing the tissue-invading function of CXCR5+CD4 cells, allowing them to infiltrate tumor tissues and carry out CD4 T cell-mediated killing and tissue destruction against antigen-specific target cells, immunotherapy can be achieved.

[0053] Example 3 Adult CD90.1 wild-type mice were immunized bilaterally in the abdomen with a dose of 30 μg / mouse MOG 35-55 mixed with complete Freund's adjuvant. Seven days later, lymph nodes were drained, and a cell suspension without erythrocytes was prepared. Figures 9-10 As shown, SLAM+CD4 cells were present in CD4crePOFUT1 conditional knockout mice, and their numbers were slightly increased; however, the CXCR5 cell population was absent.

[0054] CD4 T cell negative selection magnetic beads were used to remove non-CD4 T cell leukocytes. SLAM-positive cells in the CD4 T cell population were labeled with a fluorescently conjugated SLAM antibody; SLAM-positive cells in the CD4 T cell population were positively sorted using flow cytometry. The SLAM-positive CD4 T cells obtained by this method had a relatively high proportion of MOG35-55 specific cells.

[0055] A dose of 500,000 cells per mouse was transfused into non-CD90.1 CD4crePOFUT1 conditional knockout mice. Twenty-four hours later, mice were immunized at the base of the tail with a mixture of 30 μg / mouse MOG 35-55 complete Freund's adjuvant and pertussis toxin. CD4crePOFUT1 conditional knockout mice without SLAM-positive CD4 T cells served as negative controls, and wild-type mice as positive controls. Figures 11-14 As shown, Group 1 used CD4crePOFUT1 conditional knockout mice without SLAM-positive CD4 T cells as a negative control, with a mean symptom score of 0; Group 2 used wild-type mice as a positive control, with a maximum mean symptom score of 4; Group 3, CD4crePOFUT1 conditional knockout mice that were transfused with SLAM-positive CD4 T cells 48 hours in advance, successfully induced EAE symptoms, with a maximum mean symptom score of 4. In the central nervous system of the diseased mice, only 10% of the infiltrating CD90.1+ wild-type CD4 T cells were transfused donor cells. These donor cells, incredibly, comprised over 90% of the cells, exhibiting strong expression of cytokines and inflammatory factors (IFNγ, IL7, and GM-CSF), exhibiting typical effector cell characteristics. The remaining 90% were drained donor CD4 T cells. This result indicates that SLAM-MF1+ CD4 cells invade the mouse central nervous system from the bloodstream, becoming highly pathogenic infiltrating cells and possessing the function of draining other CD4 cells.

[0056] Based on the results of this embodiment, we can infer that: 1. MOG-specific SLAMF1+CD4 cells can invade central nervous system tissues, inducing various types of sclerosis and similar autoimmune diseases. Similar to the MOG-specific CXCR5+CD4 cells in this embodiment, other antigen-specific SLAMF1+CD4 cells can invade autologous or transplanted organ tissues, carrying out CD4 T cell-mediated killing and tissue destruction against antigen-specific target cells. For example, the kidneys in lupus nephritis; joint tissues in rheumatoid arthritis; and various organs in xenotransplantation. Therefore, they can be used as biomarkers for clinical diagnosis and treatment. 2. Utilizing the ability of SLAMF1+CD4 cells to invade tissues and draw in other immune cells, allowing them to infiltrate tumor tissues and carry out CD4 T cell-mediated killing and tissue destruction against antigen-specific target cells, immunotherapy can be achieved.

[0057] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be determined by the claims.

Claims

1. The application of protein O-fucosyltransferase POFUT1 or its encoded gene as a target in the preparation of drugs for the prevention, relief and / or treatment of autoimmune diseases and organ transplant rejection.

2. The application according to claim 1, characterized in that, The application is to prevent, alleviate, and / or treat autoimmune diseases by inhibiting the expression of the protein O-fucosyltransferase POFUT1 or its encoded gene in CD4 T cells.

3. The application according to claim 1, characterized in that, The autoimmune diseases mentioned include, but are not limited to, systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, ulcerative colitis, and rejection of allogeneic organ transplants.

4. The application according to claim 1, characterized in that, The drug includes an inhibitor that suppresses the expression of the protein O-fucosyltransferase POFUT1 in CD4 T cells.

5. The application according to claim 1, characterized in that, The application includes at least one of the following functions: (1) Inhibits the infiltration ability of SLAMF1-positive CD4 T cells; (2) Block the development of CXCR5-positive CD4 T cells.

6. The application of protein O-fucosyltransferase POFUT1 or its encoded gene as a target in the preparation of inhibitors for autoimmune diseases.

7. The application according to claim 6, characterized in that, The blocking agent can bind to the target site and inhibit the expression of the protein O-fucosyltransferase POFUT1 in CD4 T cells.

8. The use of an inhibitor of protein O-fucosyltransferase POFUT1 in the preparation of a medicament for treating diseases caused by tissue-infiltrating CD4 T cells, characterized in that, The application involves inhibiting the expression of POFUT1 or its encoded gene in CD4 T cells, blocking the development of CXCR5-positive CD4 T cells, and inhibiting the infiltration ability of SLAMF1-positive CD4 T cells.

9. The use of CXCR5-positive CD4 T cells and / or SLAMF1-positive CD4 T cells as a means of preparing drugs for the prevention, relief and / or treatment of autoimmune diseases or tumors.

10. The application according to claim 9, characterized in that, The drug prevents, alleviates, and / or treats autoimmune diseases by blocking the development of CXCR5-positive CD4 T cells and inhibiting the infiltration ability of SLAMF1-positive CD4 T cells. Alternatively, the drug may utilize CXCR5-positive and SLAMF1+CD4 T cells as tissue infiltration source cells to perform CD4 T cell-mediated killing and tissue destruction functions against antigen-specific target cells (including tumor cells), and utilize this function to treat tumors.

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