[go: up one dir, main page]

WO2023277491A1 - Composition containing immunosuppressive t cells for preventing or treating immune-related diseases - Google Patents

Composition containing immunosuppressive t cells for preventing or treating immune-related diseases Download PDF

Info

Publication number
WO2023277491A1
WO2023277491A1 PCT/KR2022/009151 KR2022009151W WO2023277491A1 WO 2023277491 A1 WO2023277491 A1 WO 2023277491A1 KR 2022009151 W KR2022009151 W KR 2022009151W WO 2023277491 A1 WO2023277491 A1 WO 2023277491A1
Authority
WO
WIPO (PCT)
Prior art keywords
cells
immune
disease
tim3
preventing
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/KR2022/009151
Other languages
French (fr)
Korean (ko)
Inventor
김정아
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Industry Academic Cooperation Foundation of Catholic University of Korea
Original Assignee
Industry Academic Cooperation Foundation of Catholic University of Korea
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from KR1020220077594A external-priority patent/KR102873199B1/en
Application filed by Industry Academic Cooperation Foundation of Catholic University of Korea filed Critical Industry Academic Cooperation Foundation of Catholic University of Korea
Publication of WO2023277491A1 publication Critical patent/WO2023277491A1/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/10—Cellular immunotherapy characterised by the cell type used
    • A61K40/11—T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/20—Cellular immunotherapy characterised by the effect or the function of the cells
    • A61K40/22—Immunosuppressive or immunotolerising
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/40—Cellular immunotherapy characterised by antigens that are targeted or presented by cells of the immune system
    • A61K40/41—Vertebrate antigens
    • A61K40/418—Antigens related to induction of tolerance to non-self
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/40—Cellular immunotherapy characterised by antigens that are targeted or presented by cells of the immune system
    • A61K40/41—Vertebrate antigens
    • A61K40/42—Cancer antigens
    • A61K40/4202—Receptors, cell surface antigens or cell surface determinants
    • A61K40/421—Immunoglobulin superfamily
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the present invention relates to a composition for preventing or treating immune-related diseases including T cells exhibiting an immunosuppressive effect, and more specifically, to preventing or treating immune-related diseases including T cells having CD3+, PD1+ and Tim3+ immune phenotypes. It relates to therapeutic compositions.
  • MSC Mesenchymal Stem Cells
  • Antigen invasion signals are transmitted by antigen presenting cells through the TCR of T cells and co-stimulatory molecules, resulting in an immune response.
  • T cells differentiate into central memory cells, whereas under continuous stimulation, T cells produce co-inhibitory molecules such as PD-1, Tim3, LAG-3, and TIGIT. expression, and the surface antigen-converted T cells block excessive immune responses and maintain a tolerance state. Therefore, if a cell therapy agent capable of suppressing an immune response by inducing T cells transformed with a surface antigen can be developed, it will be useful for various purposes in various immune diseases.
  • studies on selective inactivation of T cell surface antigens by DNA cleavage are in progress (Korean Patent Publication No. 10-2016-0029017), most studies are only on non-alloreactive immunosuppression, Studies on T cells and their use for suppressing the alloimmune response are still insignificant.
  • graft-versus-host disease is an allogeneic immune response caused by a mismatch between donor and patient histocompatibility antigens (HLA) after transplantation, with a frequency ranging from 10% to 80%. Severe GvHD (grade 3-4) with a high mortality rate is also known to occur in about 20% of patients.
  • Various immunosuppressants such as immunoglobulin, thalidomide, and cyclosporine are used for the treatment of graft-versus-host disease, but most patients die from infection or long-term The problem of deteriorating quality of life due to impaired function has emerged. Therefore, there is an urgent need to develop cell therapy agents for preventing or treating graft-versus-host disease.
  • T cells having CD3+, PD1+, and Tim3+ immune phenotypes exhibit significant immunosuppressive effects in allogeneic monocytes, and thus cells having CD3+, PD1+, and Tim3+ immunophenotypes.
  • the purpose of preventing or treating immune-related diseases was identified.
  • an object of the present invention is to provide a pharmaceutical composition for preventing or treating immune-related diseases, including T cells having CD3+, PD1+ and Tim3+ immune phenotypes.
  • an object of the present invention is to provide a pharmaceutical preparation for preventing or treating immune-related diseases, including the above composition.
  • the present invention provides a pharmaceutical composition for preventing or treating immune-related diseases, including T cells having an immune phenotype of CD3+, PD1+ and Tim3+.
  • the T cells may further have an immunophenotype of TIGIT+ or TIGIT-.
  • the composition can suppress an immune response.
  • the immune-related disease is an autoimmune disease; graft-versus-host disease; organ transplant rejection; asthma; atopy; or an acute or chronic inflammatory disease.
  • the autoimmune disease is rheumatoid arthritis, type 1 diabetes, systemic scleroderma, systemic lupus erythematosus, systemic lupus erythematosus, atopic dermatitis, psoriasis, alopecia areata, asthma, Crohn's disease, Behçetsi disease , Sjogren's syndrome, Gilia-Barre syndrome, chronic thyroiditis, multiple sclerosis, polymyositis, ankylosing spondylitis, fibromyalitis, and polyarteritis nodosa.
  • the T cells may be prepared by culturing a CD3+ cell fraction and IL-2+ in a mixture.
  • the present invention provides a method for treating immune-related diseases comprising administering the pharmaceutical composition to a subject.
  • the present invention provides a therapeutic use of the pharmaceutical composition for immune-related diseases.
  • the present invention provides a pharmaceutical preparation for preventing or treating immune-related diseases, including the above composition.
  • the pharmaceutical preparation can suppress the immune response.
  • the immune-related disease is an autoimmune disease; graft-versus-host disease; organ transplant rejection; asthma; atopy; or an acute or chronic inflammatory disease.
  • the autoimmune disease is rheumatoid arthritis, type 1 diabetes, systemic scleroderma, systemic lupus erythematosus, systemic lupus erythematosus, atopic dermatitis, psoriasis, alopecia areata, asthma, Crohn's disease, Behçetsi disease , Sjogren's syndrome, Gilia-Barre syndrome, chronic thyroiditis, multiple sclerosis, polymyositis, ankylosing spondylitis, fibromyalitis, and polyarteritis nodosa.
  • cells having immunosuppressive CD3+, PD1+, and Tim3+ immune phenotypes can be easily collected in large quantities, and since the cells have an immunosuppressive effect, they can be used as a treatment for immune-related diseases for immunosuppression. It is expected to be widely used.
  • Figure 1 shows G-CSF mobilize PBSC CD3+ lymphocytes not cultured (PBSC T cells), cultured with IL-2 (50 U/mL) for 4 days (IL-2(+) PBSC T cells), or treated with IL-2 After culturing for 4 days (IL-2(-) PBSC T cells) and analyzing each cell, Figure 1a shows IL-2(+) PBSC T cells and IL-2(-) PBSC T cells as CD14+ allogeneic monocytes, respectively.
  • MNCs monoclear cells
  • MLR mixed lymphocyte reaction
  • FIG. 2 analyzes the immunosuppressive effect of CD3+PD1+Tim3+ cells
  • FIG. 2a shows the result of CD3+PD1+ cell fractionation and the induction of an immune response (mixed lymphocyte reaction, MLR) using only PD1- cells after removing PD1+ cells.
  • MLR mixed lymphocyte reaction
  • FIG. 2b shows the results of CD3+PD1+Tim3+ cell fractionation and the induction of a mixed lymphocyte reaction (MLR) after removing Tim3+ cells.
  • MLR mixed lymphocyte reaction
  • FIG. 3 shows the result of inducing a mixed lymphocyte reaction (MLR) according to the expression of TIGIT in the CD3+PD1+Tim3+ cell fraction.
  • MLR mixed lymphocyte reaction
  • Figure 4 analyzes the immunosuppressive effect of CD3+PD1+Tim3+TIGIT- cells, and Figure 4a shows the number of CD3+PD1+Tim3+TIGIT- cells with excellent alloimmune response suppression effect in CD3+ in PBSC T cells before culture.
  • the number of CD3+PD1+Tim3+TIGIT- cells in IL-2(+) PBSC T cells and IL-2(-) PBSC T cells after culture was compared with the cell ratio 4b shows the number of CD3+PD1+Tim3+TIGIT- cells that can be obtained by collecting PBSC once in an actual clinical trial, PBSC T cells before culture, IL-2(+) PBSC T cells and IL-2(-) cells after culture. ) Results compared with PBSC T cells.
  • the present inventors confirmed that cells with CD3+, PD1+, and Tim3+ immune phenotypes exhibit significant immunosuppressive effects in allogeneic monocytes, thereby preventing or treating immune-related diseases of T cells with CD3+, PD1+, and Tim3+ immune phenotypes. identified.
  • the present invention provides a pharmaceutical composition for preventing or treating immune-related diseases, including T cells having CD3+, PD1+ and Tim3+ immune phenotypes.
  • prevention refers to any activity that suppresses or delays the onset of immune-related diseases by administration of the composition according to the present invention.
  • treatment refers to any activity that improves or beneficially changes the symptoms of an immune-related disease by administration of the composition according to the present invention.
  • immune-related disease refers to a disease caused by dysfunction of the immune system, including autoimmune disease; graft-versus-host disease; organ transplant rejection; asthma; atopy; Or it may be an acute or chronic inflammatory disease, but is not limited thereto.
  • autoimmune disease refers to a disease that exhibits a pathological response to a self antigen
  • the autoimmune disease in the present invention includes rheumatoid arthritis, type 1 diabetes, systemic scleroderma, systemic lupus erythematosus , systemic lupus erythematosus, atopic dermatitis, psoriasis, alopecia areata, asthma, Crohn's disease, Bechetsy's disease, Sjogren's syndrome, Guillain-Barre syndrome, chronic thyroiditis, multiple sclerosis, polymyositis, ankylosing spondylitis, fibromyalgia, and polyarteritis nodosa. It may be one or more selected from the group, but is not limited thereto.
  • graft-versus-host disease is a disease that occurs when transplanted lymphocytes attack the body of a patient with reduced immune function. Graft-versus-host disease in the present invention may include acute graft-versus-host disease. It may be, but is not limited thereto.
  • the T cell may have a TIGIT+ or TIGIT- immune phenotype, but is not limited thereto.
  • the composition may suppress an immune response, more preferably suppress an alloimmune response, but is not limited thereto.
  • the T cells may be prepared by culturing a CD3+ cell fraction and IL-2+ in a mixture.
  • the present inventors confirmed through specific examples that the cells of the present invention are effective in preventing or treating immune-related diseases.
  • IL-2(+) PBSC T cells expressing co-inhibitory molecules such as PD1, Tim3 and Lag3 compared to IL-2(-) PBSC T cells
  • the percentage of cells that did not proliferate in response to allo-MNCs was 16.1 ⁇ 4.7%, showing a distinct immunosuppressive effect compared to PBSC T cells (0%) before culture.
  • the immunosuppressive effect was increased 1.5 times compared to IL-2(-) PBSC T cells (11.0 ⁇ 0.1%) (see Example 2).
  • CD3+PD1+Tim3+ cells were TIGIT-, and as a result of confirming the alloimmune response according to TIGIT expression, CD3+PD1+Tim3+TIGIT- cells did not proliferate in response to allo-MNCs.
  • the percentage of cells that did not respond was 24.3 ⁇ 9.4%, which was 1.7 times higher than that of CD3+PD1+Tim3+TIGIT+ cells (14.3 ⁇ 17.0%). It was confirmed that the % was still maintained (see Example 4).
  • the T cells of the present invention can be used as cell therapy agents for the treatment of immune-related diseases by inducing immunosuppression through the expression of co-inhibitory molecules.
  • the pharmaceutical composition according to the present invention may include cells having immunophenotypes of CD3+, PD1+, and Tim3+ alone in a pharmaceutically effective amount, or may include one or more pharmaceutically acceptable carriers.
  • the pharmaceutically acceptable carrier is one commonly used in the formulation, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose , polyvinyl pyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil, but are not limited thereto.
  • lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, and the like may be further included.
  • the pharmaceutical composition of the present invention may be administered orally or parenterally (for example, intravenously, subcutaneously, intraperitoneally or topically applied) depending on the desired method, and the dosage depends on the patient's condition, body weight, and disease. Depending on the degree, drug form, administration route and time, it can be appropriately selected by those skilled in the art.
  • the pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount.
  • pharmaceutically effective amount means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level depends on the type of disease, severity, activity of the drug, and drug sensitivity, time of administration, route of administration and excretion rate, duration of treatment, factors including concomitantly used drugs, and other factors well known in the medical arts.
  • the pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered single or multiple times. Considering all of the above factors, it is important to administer an amount that can obtain the maximum effect with the minimum amount without side effects, which can be easily determined by those skilled in the art.
  • the effective amount of the pharmaceutical composition of the present invention may vary depending on the patient's age, sex, condition, body weight, absorption rate, inactivation rate and excretion rate of the active ingredient in the body, type of disease, and concomitant drugs, generally Depending on the body weight, an appropriate dose may be administered daily or every other day, or divided into 1 to 3 times a day. In addition, the dosage may be increased or decreased depending on the route of administration, gender, age, and the like.
  • the present invention provides a pharmaceutical formulation for preventing or treating immune-related diseases, including the above composition.
  • the pharmaceutical agent may suppress an immune response, and more preferably suppress an alloimmune response, but is not limited thereto.
  • the present invention provides a method for preventing or treating an immune-related disease comprising administering T cells having an immune phenotype of CD3+, PD1+, and Tim3+ to a subject in need thereof.
  • CD3+ “CD3+”, “PD1+”, “Tim3+”, “immunophenotype”, “T cell”, “immune-related disease”, “administration”, “prevention” or “treatment” may be within the aforementioned range.
  • the "individual” may be a mammal such as a rat, livestock, mouse, human, etc., and specifically may be a companion dog, racehorse, human, etc. in need of treatment for a metabolic disease, and preferably may be a human.
  • the T cells may further have an immunophenotype of TIGIT+ or TIGIT-.
  • the present invention provides the use of T cells having CD3+, PD1+ and Tim3+ immune phenotypes for the manufacture of a drug for preventing or treating immune-related diseases.
  • CD3+ “CD3+”, “PD1+”, “Tim3+”, “immunophenotype”, “T cell”, “immune-related disease”, “prevention” or “treatment” may be within the aforementioned range.
  • G-CSF G-CSF mobilized peripheral blood stem cells
  • monocytes were separated with Ficoll-Plaque plus (GE healthcare, 17-1440-02) and then MACS magnetic, MS column (Miltenyibiotec. 130-042-201) containing CD3 MicroBeads, human (miltenyibiotec. 130-050-101). ) to sort CD3+ lymphocytes.
  • CD3+ cells were loaded on a 100 ⁇ dish with 15 mL of serum-free medium containing IL-2 (50 U/ml, human IL-2 (peprotech, 200-02)) as a culture medium and cultured for 4 days. .
  • IL-2 50 U/ml, human IL-2 (peprotech, 200-02)
  • PBSC T cells G-CSF mobilize PBSC CD3+ lymphocytes without culture (PBSC T cells), with IL-2 (50 U/mL) for 4 days (IL-2(+) PBSC T cells), or without IL-2 treatment for 4 days.
  • IL-2(+) PBSC T cells Cultured for a day (IL-2(-) PBSC T cells), the level of expression of co-inhibitory molecules on the surface of each cell was confirmed.
  • IL-2(+) PBSC T cells and IL-2(-) PBSC T cells as described above were mixed with CD14+ allogeneic mononuclear cells (MNCs) and an immune response (mixed lymphocyte reaction, MLR) was induced.
  • MNCs CD14+ allogeneic mononuclear cells
  • MLR mixed lymphocyte reaction
  • Tim3+ cells were removed from the CD3+PD1+ cell group and then a mixed lymphocyte reaction (MLR) was induced. As a result, Tim3+ cells were removed from the CD3+PD1+ cell group. In this case, the effect of suppressing the allogeneic immune response was confirmed to be lost. Therefore, it was confirmed from the above results that T cells suppressing the allogeneic immune response exist in CD3+PD1+Tim3+ cells.
  • MLR mixed lymphocyte reaction
  • CD3+PD1+Tim3+ cells were TIGIT-, and as a result of confirming the alloimmune response according to TIGIT expression, CD3+PD1+Tim3+TIGIT- cells did not proliferate in response to allo-MNCs.
  • the percentage of cells that did not respond was 24.3 ⁇ 9.4%, which was 1.7 times higher than that of CD3+PD1+Tim3+TIGIT+ cells (14.3 ⁇ 17.0%). It was confirmed that the % was still maintained.
  • the number of CD3+PD1+Tim3+TIGIT-cells with excellent immunosuppressive effect was about 10 times higher in the IL-2(+) PBSC T cell group compared to PBSC 0 day (control group). Increased.
  • the number of CD3+PD1+Tim3+TIGIT- cells increased by about 6.6 times compared to that of PBSC T cells (control group).
  • CD3+PD1+Tim3+TIGIT- cells account for 15.8 ⁇ 10.2% of total CD3+ cells, which is necessary for treatment related to immunosuppressive response by collecting and culturing PBSCs once. It was found that a sufficient number of cells (a total of 1.5 ⁇ 1.1 x 10 8 CD3+PD1+Tim3+TIGIT- cells per unit weight) could be obtained.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • Immunology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Transplantation (AREA)
  • Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

The present invention relates to a composition, containing T cells exhibiting an immunosuppressive effect, for preventing or treating immune-related diseases, and more specifically, relates to a composition for preventing or treating immune-related diseases, wherein the composition contains cells having immune phenotypes of CD3+, PD1+ and Tim3+. According to the present invention, it is possible to easily collect large quantities of cells having immune phenotypes of CD3+, PD1+ and Tim3+ having an immunosuppressive effect. Due to having an immunosuppressive effect, the cells are expected to be widely used as a therapeutic agent for immune-related diseases for immunosuppression.

Description

면역 억제 T 세포를 포함하는 면역 관련 질환의 예방 또는 치료용 조성물Composition for preventing or treating immune-related diseases containing immunosuppressive T cells

본 발명은 면역억제 효과를 나타내는 T 세포를 포함하는 면역 관련 질환의 예방 또는 치료용 조성물에 관한 것으로, 보다 구체적으로는 CD3+, PD1+ 및 Tim3+의 면역 표현형을 갖는 T 세포를 포함하는 면역 관련 질환 예방 또는 치료용 조성물에 관한 것이다. The present invention relates to a composition for preventing or treating immune-related diseases including T cells exhibiting an immunosuppressive effect, and more specifically, to preventing or treating immune-related diseases including T cells having CD3+, PD1+ and Tim3+ immune phenotypes. It relates to therapeutic compositions.

면역억제용 세포 치료제에 대한 시대적 요구가 증가됨에 따라, 면역반응을 일방적으로 억제하는 면역억제제보다는 세포 간의 균형을 교정하면서 동종면역반응을 조절할 수 있는 면역세포치료가 가장 이상적으로 평가되고 있다. 하지만, 면역 억제용 세포치료제인 MSC(Mesenchymal Stem Cells)는 세포 수의 확보를 위해 여러 단계의 배양 과정이 필요하며, 이 과정에서 발생하는 활성산소로 인해 세포기능이 손상된다.As the demand for cell therapy for immunosuppression increases, immune cell therapy that can control the alloimmune response while correcting the balance between cells is evaluated as the most ideal, rather than immunosuppressive agents that unilaterally suppress the immune response. However, MSC (Mesenchymal Stem Cells), a cell therapy for immunosuppression, requires a multi-step culture process to secure the number of cells, and cell function is damaged due to active oxygen generated during this process.

항원 침입 신호는 항원 제시 세포(antigene presenting cells)가 T 세포의 TCR과 공동 자극 인자(co-stimulatory molecule)를 통해 신호를 전달하여 면역 반응이 일어난다. 면역반응이 종료되면 T 세포는 central memory cell로 분화되는 반면, 지속적인 자극이 있는 상태에서는 T 세포가 PD-1을 비롯한, Tim3, LAG-3, TIGIT 등과 같은 공동 억제 인자(co-inhibitory molecule)가 발현하게 되고, 이렇게 표면항원이 변환된 T 세포는 면역반응이 과다하게 일어나는 것을 차단하고 tolerance 상태를 유지하게 된다. 따라서, 표면항원이 변환된 T 세포를 유도하여 면역반응을 억제시킬 수 있는 세포치료제를 개발할 수 있다면 여러 면역 질환에서 다양한 용도로 유용하게 사용될 수 있을 것이다. DNA 절단(cleavage)으로 T 세포 표면항원을 선택적으로 불활성화시키는 연구가 진행 중이긴 하나(한국 공개특허 10-2016-0029017), 비-동종반응성(non-alloreactive) 면역 억제 관한 연구가 대부분일 뿐, 동종면역반응을 억제시키는 T 세포 및 이의 용도에 대해서는 아직까지 연구가 미미한 실정이다.Antigen invasion signals are transmitted by antigen presenting cells through the TCR of T cells and co-stimulatory molecules, resulting in an immune response. When the immune response is terminated, T cells differentiate into central memory cells, whereas under continuous stimulation, T cells produce co-inhibitory molecules such as PD-1, Tim3, LAG-3, and TIGIT. expression, and the surface antigen-converted T cells block excessive immune responses and maintain a tolerance state. Therefore, if a cell therapy agent capable of suppressing an immune response by inducing T cells transformed with a surface antigen can be developed, it will be useful for various purposes in various immune diseases. Although studies on selective inactivation of T cell surface antigens by DNA cleavage are in progress (Korean Patent Publication No. 10-2016-0029017), most studies are only on non-alloreactive immunosuppression, Studies on T cells and their use for suppressing the alloimmune response are still insignificant.

한편, 이식편대숙주병(graft-versus-host disease, GvHD)은 이식 후 공여자와 환자의 조직적합원(histocompativility antigens, HLA)이 불일치하여 발생하는 동종 면역반응으로, 빈도는 10%부터 80%까지 나타나며, 사망률이 높은 중증 GvHD(grade 3~4)의 경우도 약 20% 환자에서 발생하는 것으로 알려져 있다. 이식편대 숙주병의 치료를 위해 스테로이드 치료부터 면역글로불린, 탈리도마이드(thalidomide), 사이클로스포린(cyclosporine)등의 면역억제제를 다양하게 사용하고 있으나, 스테로이드에 반응이 없는 경우 대부분의 환자는 감염으로 사망하거나, 장기기능이 손상되어 삶의 질이 저하되는 문제점이 대두되고 있다. 따라서, 이식편대 숙주병의 예방 또는 치료를 위한 세포치료제의 개발이 시급한 실정이다. On the other hand, graft-versus-host disease (GvHD) is an allogeneic immune response caused by a mismatch between donor and patient histocompatibility antigens (HLA) after transplantation, with a frequency ranging from 10% to 80%. Severe GvHD (grade 3-4) with a high mortality rate is also known to occur in about 20% of patients. Various immunosuppressants such as immunoglobulin, thalidomide, and cyclosporine are used for the treatment of graft-versus-host disease, but most patients die from infection or long-term The problem of deteriorating quality of life due to impaired function has emerged. Therefore, there is an urgent need to develop cell therapy agents for preventing or treating graft-versus-host disease.

상기와 같은 문제점을 해결하기 위하여, 본 발명자들은 CD3+, PD1+ 및 Tim3+의 면역 표현형을 갖는 T 세포가 동종 단핵구에서 유의적인 면역억제 효과를 나타내는 것을 확인함으로써, CD3+, PD1+ 및 Tim3+ 의 면역 표현형을 갖는 세포의 면역 관련 질환 예방 또는 치료용도를 규명하였다. In order to solve the above problems, the present inventors confirmed that T cells having CD3+, PD1+, and Tim3+ immune phenotypes exhibit significant immunosuppressive effects in allogeneic monocytes, and thus cells having CD3+, PD1+, and Tim3+ immunophenotypes. The purpose of preventing or treating immune-related diseases was identified.

이에, 본 발명은 CD3+, PD1+ 및 Tim3+ 의 면역 표현형을 갖는 T 세포를 포함하는, 면역 관련 질환의 예방 또는 치료용 약학적 조성물을 제공하는 것을 목적으로 한다. Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing or treating immune-related diseases, including T cells having CD3+, PD1+ and Tim3+ immune phenotypes.

또한, 본 발명은 상기 조성물을 포함하는, 면역 관련 질환의 예방 또는 치료용 약학제제를 제공하는 것을 목적으로 한다.In addition, an object of the present invention is to provide a pharmaceutical preparation for preventing or treating immune-related diseases, including the above composition.

그러나 본 발명이 이루고자 하는 기술적 과제는 이상에서 언급한 과제에 제한되지 않으며, 언급되지 않은 또 다른 과제들은 아래의 기재로부터 당업자에게 명확하게 이해될 수 있을 것이다.However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problems, and other problems not mentioned will be clearly understood by those skilled in the art from the following description.

상기와 같은 목적을 달성하기 위하여 본 발명은, CD3+, PD1+ 및 Tim3+의 면역 표현형을 갖는 T 세포를 포함하는, 면역 관련 질환의 예방 또는 치료용 약학적 조성물을 제공한다.In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating immune-related diseases, including T cells having an immune phenotype of CD3+, PD1+ and Tim3+.

본 발명의 일구현예로, 상기 T 세포는 TIGIT+ 또는 TIGIT-의 면역 표현형을 더 갖는 것일 수 있다.In one embodiment of the present invention, the T cells may further have an immunophenotype of TIGIT+ or TIGIT-.

본 발명의 다른 구현예로, 상기 조성물은 면역 반응을 억제할 수 있다.In another embodiment of the present invention, the composition can suppress an immune response.

본 발명의 또 다른 구현예로, 상기 면역 관련 질환은 자가면역질환; 이식편대숙주 질환; 장기이식거부반응; 천식; 아토피; 또는 급성 또는 만성의 염증 질환일 수 있다.In another embodiment of the present invention, the immune-related disease is an autoimmune disease; graft-versus-host disease; organ transplant rejection; asthma; atopy; or an acute or chronic inflammatory disease.

본 발명의 또 다른 구현예로, 상기 자가면역질환은 류마티스성 관절염, 제1형 당뇨, 전신성 경피증, 전신 홍반성 낭창, 전신홍반루프스, 아토피 피부염, 건선, 원형탈모증, 천식, 크론씨병, 베체시병, 쇼그렌 증후군, 길리아-바레 증후군, 만성 갑상선염, 다발성 경화증, 다발성 근염, 강직성 척추염, 섬유조직염 및 결절성 다발성 동맥염으로 구성된 군으로부터 선택되는 하나 이상일 수 있다.In another embodiment of the present invention, the autoimmune disease is rheumatoid arthritis, type 1 diabetes, systemic scleroderma, systemic lupus erythematosus, systemic lupus erythematosus, atopic dermatitis, psoriasis, alopecia areata, asthma, Crohn's disease, Behçetsi disease , Sjogren's syndrome, Gilia-Barre syndrome, chronic thyroiditis, multiple sclerosis, polymyositis, ankylosing spondylitis, fibromyalitis, and polyarteritis nodosa.

본 발명의 또 다른 구현예로, 상기 T 세포는 CD3+ 세포분획과 IL-2+를 혼합 배양하여 제조될 수 있다.In another embodiment of the present invention, the T cells may be prepared by culturing a CD3+ cell fraction and IL-2+ in a mixture.

또한, 본 발명은 상기 약학적 조성물을 개체에 투여하는 단계를 포함하는 면역 관련 질환의 치료 방법을 제공한다.In addition, the present invention provides a method for treating immune-related diseases comprising administering the pharmaceutical composition to a subject.

또한, 본 발명은 상기 약학적 조성물의 면역 관련 질환의 치료 용도를 제공한다.In addition, the present invention provides a therapeutic use of the pharmaceutical composition for immune-related diseases.

또한, 본 발명은 상기 조성물을 포함하는, 면역 관련 질환의 예방 또는 치료용 약학제제를 제공한다.In addition, the present invention provides a pharmaceutical preparation for preventing or treating immune-related diseases, including the above composition.

본 발명의 일구현예로, 상기 약학제제는 면역 반응을 억제할 수 있다.In one embodiment of the present invention, the pharmaceutical preparation can suppress the immune response.

본 발명의 다른 구현예로, 상기 면역 관련 질환은 자가면역질환; 이식편대숙주 질환; 장기이식거부반응; 천식; 아토피; 또는 급성 또는 만성의 염증 질환일 수 있다.In another embodiment of the present invention, the immune-related disease is an autoimmune disease; graft-versus-host disease; organ transplant rejection; asthma; atopy; or an acute or chronic inflammatory disease.

본 발명의 또 다른 구현예로, 상기 자가면역질환은 류마티스성 관절염, 제1형 당뇨, 전신성 경피증, 전신 홍반성 낭창, 전신홍반루프스, 아토피 피부염, 건선, 원형탈모증, 천식, 크론씨병, 베체시병, 쇼그렌 증후군, 길리아-바레 증후군, 만성 갑상선염, 다발성 경화증, 다발성 근염, 강직성 척추염, 섬유조직염 및 결절성 다발성 동맥염으로 구성된 군으로부터 선택되는 하나 이상일 수 있다.In another embodiment of the present invention, the autoimmune disease is rheumatoid arthritis, type 1 diabetes, systemic scleroderma, systemic lupus erythematosus, systemic lupus erythematosus, atopic dermatitis, psoriasis, alopecia areata, asthma, Crohn's disease, Behçetsi disease , Sjogren's syndrome, Gilia-Barre syndrome, chronic thyroiditis, multiple sclerosis, polymyositis, ankylosing spondylitis, fibromyalitis, and polyarteritis nodosa.

본 발명에 의하여 면역억제 효과가 있는 CD3+, PD1+ 및 Tim3+의 면역 표현형을 갖는 세포를 대량으로 손쉽게 수집할 수 있으며, 상기 세포는 면역반응 억제 효과를 가지므로, 면역 억제를 위한 면역 관련 질환의 치료제로 널리 활용될 수 있을 것으로 기대된다.According to the present invention, cells having immunosuppressive CD3+, PD1+, and Tim3+ immune phenotypes can be easily collected in large quantities, and since the cells have an immunosuppressive effect, they can be used as a treatment for immune-related diseases for immunosuppression. It is expected to be widely used.

도 1은 G-CSF mobilize PBSC CD3+ 림프구를 배양하지 않거나(PBSC T 세포), IL-2(50U/mL)와 함께 4일간 배양(IL-2(+) PBSC T 세포)하거나, IL-2 처리 없이 4일간 배양(IL-2(-) PBSC T 세포)하고 각 세포를 분석한 것으로, 도 1a는 IL-2(+) PBSC T 세포와 IL-2(-) PBSC T 세포를 각각 CD14+ 동종 단핵구(mononuclear cells; MNCs)와 혼합하고 면역반응(mixed lymphocyte reaction, MLR)을 유도한 결과를 나타낸 것이고, 도 1b는 각 세포 표면에서 co-inhibitory molecule의 발현 정도를 확인한 것이고, 도 1c는 각 세포의 세포 분획을 비교한 결과를 나타낸 것이다.Figure 1 shows G-CSF mobilize PBSC CD3+ lymphocytes not cultured (PBSC T cells), cultured with IL-2 (50 U/mL) for 4 days (IL-2(+) PBSC T cells), or treated with IL-2 After culturing for 4 days (IL-2(-) PBSC T cells) and analyzing each cell, Figure 1a shows IL-2(+) PBSC T cells and IL-2(-) PBSC T cells as CD14+ allogeneic monocytes, respectively. (mononuclear cells; MNCs) and the result of inducing an immune response (mixed lymphocyte reaction, MLR) is shown, Figure 1b confirms the expression level of co-inhibitory molecules on the surface of each cell, It shows the result of comparing cell fractions.

도 2는 CD3+PD1+Tim3+ 세포의 면역억제 효과를 분석한 것으로, 도 2a는 CD3+PD1+ 세포 분획 결과 및 PD1+세포를 제거한 후 PD1- 세포만을 이용하여 면역반응(mixed lymphocyte reaction, MLR)을 유도한 결과를 나타낸 것이고, 도 2b는 CD3+PD1+Tim3+ 세포 분획 결과 및 Tim3+ 세포를 제거한 후 면역반응(mixed lymphocyte reaction, MLR)을 유도한 결과를 나타낸 것이다.FIG. 2 analyzes the immunosuppressive effect of CD3+PD1+Tim3+ cells, and FIG. 2a shows the result of CD3+PD1+ cell fractionation and the induction of an immune response (mixed lymphocyte reaction, MLR) using only PD1- cells after removing PD1+ cells. One result is shown, and FIG. 2b shows the results of CD3+PD1+Tim3+ cell fractionation and the induction of a mixed lymphocyte reaction (MLR) after removing Tim3+ cells.

도 3은 CD3+PD1+Tim3+ 세포 분획 내에서 TIGIT 발현 여부에 따라 면역반응(mixed lymphocyte reaction, MLR)을 유도한 결과를 나타낸 것이다.3 shows the result of inducing a mixed lymphocyte reaction (MLR) according to the expression of TIGIT in the CD3+PD1+Tim3+ cell fraction.

도 4는 CD3+PD1+Tim3+TIGIT- 세포의 면역억제 효과를 분석한 것으로, 도 4a는 동종면역반응 억제효과가 탁월한 CD3+PD1+Tim3+TIGIT- 세포 수를 배양 전의 PBSC T 세포내의 CD3+PD1+Tim3+TIGIT- 세포 수를 기준으로 배양후의 IL-2(+) PBSC T 세포 및 IL-2(-) PBSC T 세포내의 CD3+PD1+Tim3+TIGIT- 세포수와 cell ratio로 비교한 결과이고, 도 4b는 실제 임상에서 일회 PBSC를 수집하여 얻을 수 있는 CD3+PD1+Tim3+TIGIT- 세포 수를 배양 전의 PBSC T 세포, 배양 후의 IL-2(+) PBSC T 세포 및 IL-2(-) PBSC T 세포에서 비교한 결과이다.Figure 4 analyzes the immunosuppressive effect of CD3+PD1+Tim3+TIGIT- cells, and Figure 4a shows the number of CD3+PD1+Tim3+TIGIT- cells with excellent alloimmune response suppression effect in CD3+ in PBSC T cells before culture. Based on the number of PD1+Tim3+TIGIT- cells, the number of CD3+PD1+Tim3+TIGIT- cells in IL-2(+) PBSC T cells and IL-2(-) PBSC T cells after culture was compared with the cell ratio 4b shows the number of CD3+PD1+Tim3+TIGIT- cells that can be obtained by collecting PBSC once in an actual clinical trial, PBSC T cells before culture, IL-2(+) PBSC T cells and IL-2(-) cells after culture. ) Results compared with PBSC T cells.

이하 본 발명을 실시예를 통하여 보다 상세하게 설명한다. 그러나, 이들 실시예는 본 발명을 예시적으로 설명하기 위한 것으로 본 발명의 범위가 이들 실시예에 한정되는 것은 아니다. Hereinafter, the present invention will be described in more detail through examples. However, these examples are intended to illustrate the present invention by way of example, and the scope of the present invention is not limited to these examples.

본 발명자들은 CD3+, PD1+ 및 Tim3+의 면역 표현형을 갖는 세포가 동종 단핵구에서 유의적인 면역억제 효과를 나타내는 것을 확인함으로써, CD3+, PD1+ 및 Tim3+의 면역 표현형을 갖는 T 세포의 면역 관련 질환의 예방 또는 치료용도를 규명하였다. The present inventors confirmed that cells with CD3+, PD1+, and Tim3+ immune phenotypes exhibit significant immunosuppressive effects in allogeneic monocytes, thereby preventing or treating immune-related diseases of T cells with CD3+, PD1+, and Tim3+ immune phenotypes. identified.

이에, 본 발명은 CD3+, PD1+ 및 Tim3+의 면역 표현형을 갖는 T 세포를 포함하는, 면역 관련 질환의 예방 또는 치료용 약학적 조성물을 제공한다.Accordingly, the present invention provides a pharmaceutical composition for preventing or treating immune-related diseases, including T cells having CD3+, PD1+ and Tim3+ immune phenotypes.

본 발명에서 사용되는 용어, "예방"이란 본 발명에 따른 조성물의 투여에 의해 면역 관련 질환을 억제시키거나 발병을 지연시키는 모든 행위를 의미한다.As used herein, the term "prevention" refers to any activity that suppresses or delays the onset of immune-related diseases by administration of the composition according to the present invention.

본 발명에서 사용되는 용어, "치료"란 본 발명에 따른 조성물의 투여에 의해 면역 관련 질환에 대한 증세가 호전되거나 이롭게 변경되는 모든 행위를 의미한다.As used herein, the term "treatment" refers to any activity that improves or beneficially changes the symptoms of an immune-related disease by administration of the composition according to the present invention.

본 발명에서 사용되는 용어, "면역 관련 질환"은 면역계에 기능 이상으로 인해 생기는 질환을 의미하며, 자가면역질환; 이식편대숙주 질환; 장기이식거부반응; 천식; 아토피; 또는 급성 또는 만성의 염증 질환일 수 있으나, 이에 제한되는 것은 아니다.As used herein, the term "immune-related disease" refers to a disease caused by dysfunction of the immune system, including autoimmune disease; graft-versus-host disease; organ transplant rejection; asthma; atopy; Or it may be an acute or chronic inflammatory disease, but is not limited thereto.

본 발명에서 사용되는 용어, "자가면역질환"은 자가 항원에 대한 병리적 반응을 나타내는 질환을 의미하며, 본 발명에서 자가면역질환은 류마티스성 관절염, 제1형 당뇨, 전신성 경피증, 전신 홍반성 낭창, 전신홍반루프스, 아토피 피부염, 건선, 원형탈모증, 천식, 크론씨병, 베체시병, 쇼그렌 증후군, 길리아-바레 증후군, 만성 갑상선염, 다발성 경화증, 다발성 근염, 강직성 척추염, 섬유조직염 및 결절성 다발성 동맥염으로 구성된 군으로부터 선택되는 하나 이상일 수 있으나, 이에 제한되는 것은 아니다.As used herein, the term "autoimmune disease" refers to a disease that exhibits a pathological response to a self antigen, and the autoimmune disease in the present invention includes rheumatoid arthritis, type 1 diabetes, systemic scleroderma, systemic lupus erythematosus , systemic lupus erythematosus, atopic dermatitis, psoriasis, alopecia areata, asthma, Crohn's disease, Bechetsy's disease, Sjogren's syndrome, Guillain-Barre syndrome, chronic thyroiditis, multiple sclerosis, polymyositis, ankylosing spondylitis, fibromyalgia, and polyarteritis nodosa. It may be one or more selected from the group, but is not limited thereto.

본 발명에서 사용되는 용어, "이식편대숙주병"은 이식된 림프구가 면역 기능이 저하된 환자의 몸을 공격하면서 발생하는 질환으로, 본 발명에서 이식편대숙주병은 급성이식편대숙주병을 포함할 수 있으나, 이에 제한되는 것은 아니다. As used herein, the term "graft-versus-host disease" is a disease that occurs when transplanted lymphocytes attack the body of a patient with reduced immune function. Graft-versus-host disease in the present invention may include acute graft-versus-host disease. It may be, but is not limited thereto.

본 발명에서 상기 T 세포는 TIGIT+ 또는 TIGIT-의 면역 표현형을 더 갖는 것일 수 있으나, 이에 제한되는 것은 아니다.In the present invention, the T cell may have a TIGIT+ or TIGIT- immune phenotype, but is not limited thereto.

또한, 본 발명에서 상기 조성물은 면역 반응을 억제하는 것일 수 있으며, 보다 바람직하게는 동종 면역 반응을 억제하는 것일 수 있으나 이에 제한되는 것은 아니다.In addition, in the present invention, the composition may suppress an immune response, more preferably suppress an alloimmune response, but is not limited thereto.

또한, 상기 T 세포는 CD3+ 세포분획과 IL-2+를 혼합 배양하여 제조된 것일 수 있다.In addition, the T cells may be prepared by culturing a CD3+ cell fraction and IL-2+ in a mixture.

본 발명자들은 구체적인 실시예를 통해 본 발명의 세포가 면역 관련 질환의 예방 또는 치료에 효과적임을 확인하였다.The present inventors confirmed through specific examples that the cells of the present invention are effective in preventing or treating immune-related diseases.

본 발명의 일실시예에서는 IL-2(-) PBSC T 세포와 비교하여 PD1, Tim3 및 Lag3 등의 co-inhibitory molecule이 발현된 IL-2(+) PBSC T 세포의 동종면역반응을 확인한 결과, IL-2(+) PBSC T 세포의 경우, allo-MNCs에 대해 반응하여 증식하지 않는 세포의 비율이 16.1±4.7%로 나타나 배양전의 PBSC T세포 (0%)에 비해 뚜렷한 면역억제 효과를 보였으며. IL-2(-) PBSC T 세포(11.0±0.1%)에 비해서도 1.5배 면역억제 효과가 증가되는 것을 확인하였다(실시예 2 참조).In one embodiment of the present invention, as a result of confirming the alloimmune response of IL-2(+) PBSC T cells expressing co-inhibitory molecules such as PD1, Tim3 and Lag3 compared to IL-2(-) PBSC T cells, In the case of IL-2(+) PBSC T cells, the percentage of cells that did not proliferate in response to allo-MNCs was 16.1±4.7%, showing a distinct immunosuppressive effect compared to PBSC T cells (0%) before culture. . It was confirmed that the immunosuppressive effect was increased 1.5 times compared to IL-2(-) PBSC T cells (11.0±0.1%) (see Example 2).

본 발명의 다른 실시예에서는 CD3+PD1+ 세포 내에서 PD1 또는 Tim3를 제거한 후 면역반응(mixed lymphocyte reaction, MLR)을 유도한 결과, 동종 면역반응 억제되는 효과가 소실되는 결과가 나타나, CD3+PD1+Tim3+ 세포 내에 면역반응을 억제하는 T 세포가 존재한다는 것을 확인하였다(실시예 3 참조).In another embodiment of the present invention, as a result of inducing a mixed lymphocyte reaction (MLR) after removing PD1 or Tim3 from CD3+PD1+ cells, the effect of suppressing the allogeneic immune response is lost, resulting in CD3+PD1+ It was confirmed that T cells suppressing the immune response exist in Tim3+ cells (see Example 3).

이에 더하여, 상기 CD3+PD1+Tim3+ 세포는 대부분이 TIGIT-이었으며, TIGIT 발현 여부에 따라 동종면역반응을 확인한 결과, CD3+PD1+Tim3+TIGIT-세포의 경우, allo-MNCs에 대해 반응하여 증식하지 않는 세포의 비율이 24.3±9.4%로 나타나 CD3+PD1+Tim3+TIGIT+세포(14.3±17.0%)에 비해 1.7배 증가되었으나, CD3+PD1+Tim3+TIGIT+세포에서도 동종면역반응억제 효과가 14.3±17.0%로 여전히 유지되는 것을 확인하였다(실시예 4 참조).In addition, most of the CD3+PD1+Tim3+ cells were TIGIT-, and as a result of confirming the alloimmune response according to TIGIT expression, CD3+PD1+Tim3+TIGIT- cells did not proliferate in response to allo-MNCs. The percentage of cells that did not respond was 24.3±9.4%, which was 1.7 times higher than that of CD3+PD1+Tim3+TIGIT+ cells (14.3±17.0%). It was confirmed that the % was still maintained (see Example 4).

상기 실시예의 결과로부터 본 발명의 T 세포는 공동 억제 인자(co-inhibitory molecule)의 발현을 통해 면역억제를 유도함으로써 면역 관련 질환의 치료용 세포치료제로 활용될 수 있을 것으로 기대된다.From the results of the above examples, it is expected that the T cells of the present invention can be used as cell therapy agents for the treatment of immune-related diseases by inducing immunosuppression through the expression of co-inhibitory molecules.

본 발명에 따른 약학적 조성물은 약학적으로 유효한 양의 CD3+, PD1+ 및 Tim3+의 면역 표현형을 갖는 세포를 단독으로 포함하거나 하나 이상의 약학적으로 허용되는 담체를 포함할 수 있다. 이때, 약학적으로 허용되는 담체는 제제 시에 통상적으로 이용되는 것으로서, 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아고무, 인산칼슘, 알기네이트, 젤라틴, 규산 칼슘, 미세 결정성 셀룰로스, 폴리비닐 피로리돈, 셀룰로스, 물, 시럽, 메틸 셀룰로스, 메틸히드록시벤조에이트, 프로필 히드록시벤조에이트, 활석, 스테아르산 마그네슘 및 미네랄 오일 등을 포함하나, 이에 한정되는 것은 아니다. 또한, 상기 성분들 이외에 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제, 보존제 등을 추가로 포함할 수 있다.The pharmaceutical composition according to the present invention may include cells having immunophenotypes of CD3+, PD1+, and Tim3+ alone in a pharmaceutically effective amount, or may include one or more pharmaceutically acceptable carriers. At this time, the pharmaceutically acceptable carrier is one commonly used in the formulation, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose , polyvinyl pyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil, but are not limited thereto. In addition to the above components, lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, and the like may be further included.

본 발명의 약학적 조성물은 목적하는 방법에 따라 경구 투여하거나 비경구투여 (예를 들어, 정맥 내, 피하, 복강 내 또는 국소에 적용)할 수 있으며, 투여량은 환축의 상태 및 체중, 질병의 정도, 약물형태, 투여경로 및 시간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다.The pharmaceutical composition of the present invention may be administered orally or parenterally (for example, intravenously, subcutaneously, intraperitoneally or topically applied) depending on the desired method, and the dosage depends on the patient's condition, body weight, and disease. Depending on the degree, drug form, administration route and time, it can be appropriately selected by those skilled in the art.

본 발명의 약학적 조성물은 약학적으로 유효한 양으로 투여한다. 본 발명에 있어서 "약학적으로 유효한 양"은 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분한 양을 의미하며, 유효용량 수준은 질환의 종류, 중증도, 약물의 활성, 약물에 대한 민감도, 투여 시간, 투여 경로 및 배출 비율, 치료 기간, 동시 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 알려진 요소에 따라 결정될 수 있다. 본 발명에 따른 약학적 조성물은 개별 치료제로 투여하거나 다른 치료제와 병용하여 투여될 수 있고 종래의 치료제와는 순차적 또는 동시에 투여될 수 있으며, 단일 또는 다중 투여될 수 있다. 상기 요소들을 모두 고려하여 부작용 없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하며, 이는 당업자에 의해 용이하게 결정될 수 있다.The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level depends on the type of disease, severity, activity of the drug, and drug sensitivity, time of administration, route of administration and excretion rate, duration of treatment, factors including concomitantly used drugs, and other factors well known in the medical arts. The pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered single or multiple times. Considering all of the above factors, it is important to administer an amount that can obtain the maximum effect with the minimum amount without side effects, which can be easily determined by those skilled in the art.

구체적으로 본 발명의 약학적 조성물의 유효량은 환자의 연령, 성별, 상태, 체중, 체내에 활성 성분의 흡수도, 불활성율 및 배설속도, 질병종류, 병용되는 약물에 따라 달라질 수 있으며, 일반적으로는 체중에 따라 적정용량을 매일 또는 격일 투여하거나, 1일 1 내지 3회로 나누어 투여할 수 있다. 또한, 투여 경로, 성별, 연령 등에 따라서 투여량은 증감 될 수 있다.Specifically, the effective amount of the pharmaceutical composition of the present invention may vary depending on the patient's age, sex, condition, body weight, absorption rate, inactivation rate and excretion rate of the active ingredient in the body, type of disease, and concomitant drugs, generally Depending on the body weight, an appropriate dose may be administered daily or every other day, or divided into 1 to 3 times a day. In addition, the dosage may be increased or decreased depending on the route of administration, gender, age, and the like.

또한, 본 발명의 다른 양태로서, 본 발명은 상기 조성물을 포함하는, 면역 관련 질환의 예방 또는 치료용 약학제제를 제공한다.In addition, as another aspect of the present invention, the present invention provides a pharmaceutical formulation for preventing or treating immune-related diseases, including the above composition.

본 발명에서 상기 약학제제는 면역 반응을 억제하는 것 일수 있으며, 보다 바람직하게는 동종 면역 반응을 억제하는 것일 수 있으나, 이에 제한되는 것은 아니다.In the present invention, the pharmaceutical agent may suppress an immune response, and more preferably suppress an alloimmune response, but is not limited thereto.

본 발명의 또 다른 양태로서, 본 발명은 CD3+, PD1+ 및 Tim3+의 면역 표현형을 갖는 T 세포를 이를 필요로 하는 개체에게 투여하는 단계를 포함하는 면역 관련 질환의 예방 또는 치료 방법을 제공한다.As another aspect of the present invention, the present invention provides a method for preventing or treating an immune-related disease comprising administering T cells having an immune phenotype of CD3+, PD1+, and Tim3+ to a subject in need thereof.

상기 "CD3+", "PD1+", "Tim3+", "면역 표현형", "T 세포", "면역 관련 질환", "투여", "예방" 또는 "치료" 등은 전술한 범위 내일 수 있다.The “CD3+”, “PD1+”, “Tim3+”, “immunophenotype”, “T cell”, “immune-related disease”, “administration”, “prevention” or “treatment” may be within the aforementioned range.

본 발명에서 "개체"는 쥐, 가축, 생쥐, 인간 등 포유류일 수 있으며, 구체적으로 대사성 질환의 치료가 필요한 반려견, 경주마, 인간 등일 수 있고, 바람직하게는 인간일 수 있다.In the present invention, the "individual" may be a mammal such as a rat, livestock, mouse, human, etc., and specifically may be a companion dog, racehorse, human, etc. in need of treatment for a metabolic disease, and preferably may be a human.

또한, 상기 방법에 있어서, 상기 T 세포는 TIGIT+ 또는 TIGIT-의 면역 표현형을 더 갖는 것일 수 있다.In addition, in the above method, the T cells may further have an immunophenotype of TIGIT+ or TIGIT-.

본 발명의 또 다른 양태로서, 본 발명은 면역 관련 질환의 예방 또는 치료용 약제의 제조를 위한 CD3+, PD1+ 및 Tim3+의 면역 표현형을 갖는 T 세포의 용도를 제공한다.As another aspect of the present invention, the present invention provides the use of T cells having CD3+, PD1+ and Tim3+ immune phenotypes for the manufacture of a drug for preventing or treating immune-related diseases.

상기 "CD3+", "PD1+", "Tim3+", "면역 표현형", "T 세포", "면역 관련 질환", "예방" 또는 "치료" 등은 전술한 범위 내일 수 있다.The “CD3+”, “PD1+”, “Tim3+”, “immunophenotype”, “T cell”, “immune-related disease”, “prevention” or “treatment” may be within the aforementioned range.

이하 본 발명을 실시예를 통하여 보다 상세하게 설명한다. 그러나 이들 실시예는 본 발명을 예시적으로 설명하기 위한 것으로 본 발명의 범위가 이들 실시예에 한정되는 것은 아니다.Hereinafter, the present invention will be described in more detail through examples. However, these examples are intended to illustrate the present invention by way of example, and the scope of the present invention is not limited to these examples.

[실시예][Example]

실시예 1. 세포 수집 및 증식 방법Example 1. Cell collection and propagation method

말초 조혈모세포 공여자에게 G-CSF(neutrogin; 10 ㎍/kg)를 5일간 피하 주사한 뒤 COBE spectra를 이용하여 G-CSF mobilized peripheral blood stem cell(PBSC)을 수집한다. 이후 Ficoll-Plaque plus (GE healthcare, 17-1440-02)로 단핵구를 분리한 뒤 CD3 MicroBeads, human(miltenyibiotec. 130-050-101)이 포함된 MACS magnetic, MS column(Miltenyibiotec. 130-042-201)을 이용하여, CD3+ 림프구를 sorting 한다. 이후, IL-2(50 U/ml, human IL-2(peprotech, 200-02)가 포함된 무혈청 배지 15 mL를 배양액으로 100π dish에 0.5 X 107개의 CD3+ 세포를 로딩하여 4일간 배양하였다.Peripheral hematopoietic stem cell donors were subcutaneously injected with G-CSF (neutrogin; 10 μg/kg) for 5 days, and then G-CSF mobilized peripheral blood stem cells (PBSC) were collected using COBE spectra. Afterwards, monocytes were separated with Ficoll-Plaque plus (GE healthcare, 17-1440-02) and then MACS magnetic, MS column (Miltenyibiotec. 130-042-201) containing CD3 MicroBeads, human (miltenyibiotec. 130-050-101). ) to sort CD3+ lymphocytes. Thereafter, 0.5 X 10 7 CD3+ cells were loaded on a 100π dish with 15 mL of serum-free medium containing IL-2 (50 U/ml, human IL-2 (peprotech, 200-02)) as a culture medium and cultured for 4 days. .

실시예 2. 증식된 세포의 표현형 및 면역 억제 효과 확인Example 2. Verification of phenotype and immunosuppressive effect of proliferated cells

G-CSF mobilize PBSC CD3+ 림프구를 배양하지 않거나(PBSC T 세포), IL-2(50 U/mL)와 함께 4일간 배양(IL-2(+) PBSC T 세포)하거나, IL-2 처리 없이 4일간 배양(IL-2(-) PBSC T 세포)하고, 각 세포 표면에서 co-inhibitory molecule의 발현 정도를 확인하였다. G-CSF mobilize PBSC CD3+ lymphocytes without culture (PBSC T cells), with IL-2 (50 U/mL) for 4 days (IL-2(+) PBSC T cells), or without IL-2 treatment for 4 days. Cultured for a day (IL-2(-) PBSC T cells), the level of expression of co-inhibitory molecules on the surface of each cell was confirmed.

그 결과, 상기와 동일한 IL-2(+) PBSC T 세포와 IL-2(-) PBSC T 세포를 CD14+ 동종 단핵구(mononuclear cells; MNCs)와 혼합하고 면역반응(mixed lymphocyte reaction, MLR)을 유도한 결과, 도 1a에 나타낸 바와 같이 IL-2(+) PBSC T 세포의 경우, allo-MNCs에 대해 반응하여 증식하지 않는 세포의 비율이 16.1±4.7 %로 나타나 배양전의 PBSC T세포 (0%)에 비해 뚜렷한 면역억제 효과를 보였으며. IL-2(-) PBSC T 세포(11.0±0.1%)에 비해서도 1.5배 면역억제 효과가 증가되는 것을 확인하였다.As a result, the same IL-2(+) PBSC T cells and IL-2(-) PBSC T cells as described above were mixed with CD14+ allogeneic mononuclear cells (MNCs) and an immune response (mixed lymphocyte reaction, MLR) was induced. As a result, as shown in FIG. 1a, in the case of IL-2(+) PBSC T cells, the proportion of cells that did not proliferate in response to allo-MNCs was 16.1±4.7%, compared to PBSC T cells (0%) before culture. showed a marked immunosuppressive effect. It was confirmed that the immunosuppressive effect was increased 1.5 times compared to IL-2(-) PBSC T cells (11.0±0.1%).

이에 더하여, 도 1b 및 1c에 나타낸 바와 같이 IL-2(50 U/mL)와 함께 4일간 배양된 PBSC T 세포의 경우, IL-2를 처리하지 않고 배양된 PBSC T 세포 및 배양되지 않은 PBSC에 비해 TIM3, TIGIT의 발현이 현저하게 증가되는 것을 확인하였다.In addition, in the case of PBSC T cells cultured for 4 days with IL-2 (50 U/mL), as shown in Figures 1b and 1c, PBSC T cells cultured without IL-2 treatment and uncultured PBSC In comparison, it was confirmed that the expression of TIM3 and TIGIT was significantly increased.

실시예 3. CD3+PD1+Tim3+ 세포의 면역억제 효과 확인Example 3. Confirmation of immunosuppressive effect of CD3+PD1+Tim3+ cells

G-CSF mobilize PBSC CD3+ 림프구를 IL-2(50U/mL)와 함께 4일간 배양(IL-2(+) cultured PBSC T cells)한 세포에서 PD1+ 세포의 비율을 확인한 결과, 도 2a 좌측에 나타낸 바와 같이 PD1+ 세포의 비율은 34.2±21.1%로 나타났으며, CD3+PD1+ 세포 내에서 PD1+세포를 제거한 후 PD1- 세포만을 이용하여 면역반응(mixed lymphocyte reaction, MLR)을 유도한 결과, 도 2a 우측에 나타낸 바와 같이 거의 대부분의 세포가 allo-MNC에 반응하여 증식하는 것을 확인하였다. 상기 결과로부터, 면역반응을 억제하는 T 세포가 CD3+PD1+ 세포군 중에 존재하는 것을 확인하였다.As a result of confirming the ratio of PD1+ cells in cells cultured with G-CSF mobilized PBSC CD3+ lymphocytes with IL-2 (50 U/mL) for 4 days (IL-2(+) cultured PBSC T cells), as shown in the left side of FIG. 2a As shown, the percentage of PD1+ cells was 34.2±21.1%, and as a result of inducing a mixed lymphocyte reaction (MLR) using only PD1- cells after removing PD1+ cells from CD3+PD1+ cells, the right side of FIG. 2a As shown, it was confirmed that most of the cells proliferated in response to allo-MNC. From the above results, it was confirmed that T cells suppressing the immune response were present in the CD3+PD1+ cell population.

또한, 도 2b 좌측에 나타낸 바와 같이 전체 IL-2(+) PBSC T 세포에서 CD3+PD1+Tim3+ 세포는 20.0±10.5%로 존재하는 것을 확인하였다.In addition, as shown in the left side of FIG. 2B , it was confirmed that 20.0±10.5% of CD3+PD1+Tim3+ cells were present in total IL-2(+) PBSC T cells.

이에 더하여, Tim3 발현 여부가 면역억제에 영향을 미치는지 확인하기 위해, CD3+PD1+ 세포군에서 Tim3+ 세포를 제거한 후 면역반응(mixed lymphocyte reaction, MLR)을 유도한 결과, CD3+PD1+ 세포군에서 Tim3+ 세포를 제거하는 경우 동종 면역반응이 억제되는 효과가 소실되는 결과를 확인하였다. 따라서, 상기 결과로부터 CD3+PD1+Tim3+ 세포 내에 동종 면역반응을 억제하는 T 세포가 존재한다는 것을 확인하였다.In addition, to confirm whether the expression of Tim3 affects immunosuppression, Tim3+ cells were removed from the CD3+PD1+ cell group and then a mixed lymphocyte reaction (MLR) was induced. As a result, Tim3+ cells were removed from the CD3+PD1+ cell group. In this case, the effect of suppressing the allogeneic immune response was confirmed to be lost. Therefore, it was confirmed from the above results that T cells suppressing the allogeneic immune response exist in CD3+PD1+Tim3+ cells.

실시예 4. CD3+PD1+Tim3+TIGIT- 세포의 면역억제 효과 확인Example 4. Confirmation of immunosuppressive effect of CD3+PD1+Tim3+TIGIT- cells

상기 실시예 2b에 개시된 CD3+PD1+Tim3+ 세포(20.0±10.5%)를 분석한 결과, TIGIT의 경우 도 3 좌측에 나타낸 바와 같이 전체 세포 중 CD3+PD1+Tim3+TIGIT-세포가 15.8±10.2%가 TIGIT-를, 4.0±1.9%가 TIGIT+를 발현하는 것을 확인하였다. CD3+PD1+Tim3+ 세포 중 약 80%가량의 세포가 TIGIT-였다.As a result of analyzing the CD3+PD1+Tim3+ cells (20.0±10.5%) described in Example 2b, in the case of TIGIT, 15.8±10.2% of the total cells were CD3+PD1+Tim3+TIGIT- cells, as shown in the left side of FIG. It was confirmed that expressed TIGIT- and 4.0±1.9% expressed TIGIT+. About 80% of the CD3+PD1+Tim3+ cells were TIGIT-.

이에 더하여, 상기 CD3+PD1+Tim3+ 세포는 대부분이 TIGIT-이었으며, TIGIT 발현 여부에 따라 동종면역반응을 확인한 결과, CD3+PD1+Tim3+TIGIT-세포의 경우, allo-MNCs에 대해 반응하여 증식하지 않는 세포의 비율이 24.3±9.4%로 나타나 CD3+PD1+Tim3+TIGIT+세포(14.3±17.0%)에 비해 1.7배 증가되었으나, CD3+PD1+Tim3+TIGIT+세포에서도 동종면역반응억제 효과가 14.3±17.0%로 여전히 유지되는 것을 확인하였다.In addition, most of the CD3+PD1+Tim3+ cells were TIGIT-, and as a result of confirming the alloimmune response according to TIGIT expression, CD3+PD1+Tim3+TIGIT- cells did not proliferate in response to allo-MNCs. The percentage of cells that did not respond was 24.3±9.4%, which was 1.7 times higher than that of CD3+PD1+Tim3+TIGIT+ cells (14.3±17.0%). It was confirmed that the % was still maintained.

나아가, 본 발명의 도 4a에 나타낸 바와 같이, 면역억제 효과가 우수한 CD3+PD1+Tim3+TIGIT-세포 수는 PBSC 0 day (대조군)에 비해 IL-2(+) PBSC T 세포군내에서는 약 10배 증가된다. 이에 비해 IL-2(-) PBSC T 세포 군에서는 PBSC T 세포(대조군)에 비해 CD3+PD1+Tim3+TIGIT-세포 수가 약 6.6배 증가된다. Furthermore, as shown in FIG. 4a of the present invention, the number of CD3+PD1+Tim3+TIGIT-cells with excellent immunosuppressive effect was about 10 times higher in the IL-2(+) PBSC T cell group compared to PBSC 0 day (control group). Increased. In contrast, in the IL-2(-) PBSC T cell group, the number of CD3+PD1+Tim3+TIGIT- cells increased by about 6.6 times compared to that of PBSC T cells (control group).

또한, 본 발명의 도 4b에 나타낸 바와 같이, CD3+PD1+Tim3+TIGIT-세포는 전체 CD3+ 세포 중 15.8±10.2%로 나타나, PBSC를 한번 수집하고 배양하는 방법으로 면역억제 반응과 관련된 치료에 필요한 충분한 양(단위 체중 당 총 1.5±1.1 x 108 CD3+PD1+Tim3+TIGIT- cells)의 세포 수를 확보할 수 있음을 알 수 있었다.In addition, as shown in FIG. 4b of the present invention, CD3+PD1+Tim3+TIGIT- cells account for 15.8±10.2% of total CD3+ cells, which is necessary for treatment related to immunosuppressive response by collecting and culturing PBSCs once. It was found that a sufficient number of cells (a total of 1.5±1.1 x 10 8 CD3+PD1+Tim3+TIGIT- cells per unit weight) could be obtained.

상기 실시예 결과들로부터 CD3+ 세포분획을 IL-2+와 혼합하여 배양함으로써 동종 면역 반응을 억제하는 능력이 탁월한 충분한 수의 CD3+PD1+Tim3+TIGIT-세포를 획득할 수 있으며, 배양과정을 통하여 co-inhibitory molecule의 발현을 획득한 CD3+림프구는 동종면역 반응을 억제함으로써 이식편대숙주병을 치료하기 위한 세포치료제로 활용될 수 있을 것으로 기대된다. From the results of the above examples, by culturing the CD3+ cell fraction mixed with IL-2+, it is possible to obtain a sufficient number of CD3+PD1+Tim3+TIGIT-cells excellent in suppressing the alloimmune response, and through the culture process CD3+ lymphocytes that have acquired co-inhibitory molecule expression are expected to be used as cell therapy agents to treat graft-versus-host disease by suppressing the alloimmune response.

전술한 본 발명의 설명은 예시를 위한 것이며, 본 발명이 속하는 기술분야의 통상의 지식을 가진 자는 본 발명의 기술적 사상이나 필수적인 특징을 변경하지 않고서 다른 구체적인 형태로 쉽게 변형이 가능하다는 것을 이해할 수 있을 것이다. 그러므로 이상에서 기술한 실시예들은 모든 면에서 예시적인 것이며 한정적이 아닌 것으로 이해해야만 한다.The above description of the present invention is for illustrative purposes, and those skilled in the art can understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, the embodiments described above should be understood as illustrative in all respects and not limiting.

Claims (12)

CD3+, PD1+ 및 Tim3+의 면역 표현형을 갖는 T 세포를 포함하는, 면역 관련 질환의 예방 또는 치료용 약학적 조성물.A pharmaceutical composition for preventing or treating immune-related diseases, comprising T cells having immunophenotypes of CD3+, PD1+ and Tim3+. 제1항에 있어서,According to claim 1, 상기 T 세포는 TIGIT+ 또는 TIGIT-의 면역 표현형을 더 갖는 것을 특징으로 하는, 약학적 조성물.Characterized in that the T cells further have an immunophenotype of TIGIT+ or TIGIT-, the pharmaceutical composition. 제1항 또는 제2항에 있어서,According to claim 1 or 2, 상기 조성물은 면역 반응을 억제하는 것을 특징으로 하는, 약학적 조성물.The pharmaceutical composition, characterized in that the composition inhibits the immune response. 제1항 또는 제2항에 있어서,According to claim 1 or 2, 상기 면역 관련 질환은 자가면역질환; 이식편대숙주 질환; 장기이식거부반응; 천식; 아토피; 또는 급성 또는 만성의 염증 질환인 것을 특징으로 하는, 약학적 조성물.The immune-related diseases include autoimmune diseases; graft-versus-host disease; organ transplant rejection; asthma; atopy; Or characterized in that the acute or chronic inflammatory disease, the pharmaceutical composition. 제1항 또는 제2항에 있어서,According to claim 1 or 2, 상기 자가면역질환은 류마티스성 관절염, 제1형 당뇨, 전신성 경피증, 전신 홍반성 낭창, 전신홍반루프스, 아토피 피부염, 건선, 원형탈모증, 천식, 크론씨병, 베체시병, 쇼그렌 증후군, 길리아-바레 증후군, 만성 갑상선염, 다발성 경화증, 다발성 근염, 강직성 척추염, 섬유조직염 및 결절성 다발성 동맥염으로 구성된 군으로부터 선택되는 하나 이상인 것을 특징으로 하는, 약학적 조성물.The autoimmune disease is rheumatoid arthritis, type 1 diabetes, systemic scleroderma, systemic lupus erythematosus, systemic lupus erythematosus, atopic dermatitis, psoriasis, alopecia areata, asthma, Crohn's disease, Bechetsy's disease, Sjogren's syndrome, Gilia-Barre syndrome , Chronic thyroiditis, multiple sclerosis, polymyositis, ankylosing spondylitis, fibrous tissue and polyarteritis nodosa, characterized in that at least one selected from the group consisting of, a pharmaceutical composition. 제1항 또는 제2항에 있어서,According to claim 1 or 2, 상기 T 세포는 CD3+ 세포분획과 IL-2+를 혼합 배양하여 제조된 것을 특징으로 하는, 약학적 조성물.The T cells are characterized in that prepared by mixing the CD3 + cell fraction and IL-2 + culture, the pharmaceutical composition. 제1항 또는 제2항의 조성물을 포함하는, 면역 관련 질환의 예방 또는 치료용 약학제제.A pharmaceutical preparation for preventing or treating immune-related diseases, comprising the composition of claim 1 or 2. 제7항에 있어서,According to claim 7, 상기 약학제제는 면역 반응을 억제하는 것을 특징으로 하는, 약학제제.The pharmaceutical formulation is characterized in that for suppressing the immune response, pharmaceutical formulations. 제7항에 있어서,According to claim 7, 상기 면역 관련 질환은 자가면역질환; 이식편대숙주 질환; 장기이식거부반응; 천식; 아토피; 또는 급성 또는 만성의 염증 질환인 것을 특징으로 하는, 약학제제. The immune-related diseases include autoimmune diseases; graft-versus-host disease; organ transplant rejection; asthma; atopy; Or characterized in that the acute or chronic inflammatory disease, pharmaceutical preparations. 제7항에 있어서,According to claim 7, 상기 자가면역질환은 류마티스성 관절염, 제1형 당뇨, 전신성 경피증, 전신 홍반성 낭창, 전신홍반루프스, 아토피 피부염, 건선, 원형탈모증, 천식, 크론씨병, 베체시병, 쇼그렌 증후군, 길리아-바레 증후군, 만성 갑상선염, 다발성 경화증, 다발성 근염, 강직성 척추염, 섬유조직염 및 결절성 다발성 동맥염으로 구성된 군으로부터 선택되는 하나 이상인 것을 특징으로 하는, 약학제제.The autoimmune disease is rheumatoid arthritis, type 1 diabetes, systemic scleroderma, systemic lupus erythematosus, systemic lupus erythematosus, atopic dermatitis, psoriasis, alopecia areata, asthma, Crohn's disease, Bechetsy's disease, Sjogren's syndrome, Gilia-Barre syndrome , Chronic thyroiditis, multiple sclerosis, polymyositis, ankylosing spondylitis, fibromyalgia and polyarteritis nodosa, characterized in that at least one selected from the group consisting of, pharmaceutical preparations. CD3+, PD1+ 및 Tim3+의 면역 표현형을 갖는 T 세포를 이를 필요로 하는 개체에게 투여하는 단계를 포함하는 면역 관련 질환의 예방 또는 치료 방법.A method for preventing or treating an immune-related disease comprising administering T cells having an immune phenotype of CD3+, PD1+, and Tim3+ to a subject in need thereof. 면역 관련 질환의 예방 또는 치료용 약제의 제조를 위한 CD3+, PD1+ 및 Tim3+의 면역 표현형을 갖는 T 세포의 용도.Use of T cells having CD3+, PD1+ and Tim3+ immune phenotypes for the manufacture of a drug for preventing or treating an immune-related disease.
PCT/KR2022/009151 2021-06-29 2022-06-27 Composition containing immunosuppressive t cells for preventing or treating immune-related diseases Ceased WO2023277491A1 (en)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
KR10-2021-0084634 2021-06-29
KR20210084634 2021-06-29
KR10-2022-0077594 2022-06-24
KR1020220077594A KR102873199B1 (en) 2021-06-29 2022-06-24 Composition for preventing or treating Immune Related Disease comprising immune Inhibitory T cells

Publications (1)

Publication Number Publication Date
WO2023277491A1 true WO2023277491A1 (en) 2023-01-05

Family

ID=84692916

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2022/009151 Ceased WO2023277491A1 (en) 2021-06-29 2022-06-27 Composition containing immunosuppressive t cells for preventing or treating immune-related diseases

Country Status (1)

Country Link
WO (1) WO2023277491A1 (en)

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20130110283A (en) * 2012-03-29 2013-10-10 가톨릭대학교 산학협력단 Composition for treating immune disorders comprising regulatory t cells
KR102107862B1 (en) * 2011-07-01 2020-05-07 베크만 컬터, 인코포레이티드 Regulatory t cells and methods of identifying and isolating them using cd6-expression or the combination of cd4, cd25 and cd127
KR102081418B1 (en) * 2019-09-24 2020-05-26 주식회사 이뮤니스바이오 A medium composition of regulatory t cells derived from peripheral blood mononuclear cells and cultivation method using the same
US20200163994A1 (en) * 2017-05-12 2020-05-28 University Of Massachusetts Ex vivo method of generating super regulatory t cells for the prevention of autoimmune disease
WO2021028579A1 (en) * 2019-08-14 2021-02-18 Nekonal S.A.R.L. Method for producing engineered tirc7 specific t-regulatory cells (tregs)

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR102107862B1 (en) * 2011-07-01 2020-05-07 베크만 컬터, 인코포레이티드 Regulatory t cells and methods of identifying and isolating them using cd6-expression or the combination of cd4, cd25 and cd127
KR20130110283A (en) * 2012-03-29 2013-10-10 가톨릭대학교 산학협력단 Composition for treating immune disorders comprising regulatory t cells
US20200163994A1 (en) * 2017-05-12 2020-05-28 University Of Massachusetts Ex vivo method of generating super regulatory t cells for the prevention of autoimmune disease
WO2021028579A1 (en) * 2019-08-14 2021-02-18 Nekonal S.A.R.L. Method for producing engineered tirc7 specific t-regulatory cells (tregs)
KR102081418B1 (en) * 2019-09-24 2020-05-26 주식회사 이뮤니스바이오 A medium composition of regulatory t cells derived from peripheral blood mononuclear cells and cultivation method using the same

Similar Documents

Publication Publication Date Title
Kuntz et al. The cellular basis of the impaired autologous mixed lymphocyte reaction in patients with systemic lupus erythematosus.
Saltini et al. Maintenance of alveolitis in patients with chronic beryllium disease by beryllium-specific helper T cells
DE69231927T2 (en) METHOD FOR TREATING SYSTEMIC LUPUS ERYTHEMATODES
Snowden et al. Development of psoriasis after syngeneic bone marrow transplant from psoriatic donor: further evidence for adoptive autoimmunity
Hafler et al. Oligoclonal T lymphocytes in the cerebrospinal fluid of patients with multiple sclerosis.
Morimoto et al. Direct demonstration of the human suppressor inducer subset by anti-T cell antibodies.
Herrmann et al. In vitro regulation of human hematopoiesis by natural killer cells: analysis at a clonal level
Esquifino et al. Immune response after experimental allergic encephalomyelitis in rats subjected to calorie restriction
WO2019151840A1 (en) Pharmaceutical composition comprising isolated mitochondria for prevention or treatment of rheumatoid arthritis
Ford et al. Association of an interleukin abnormality with the T cell defect in Hodgkin's disease
Delfraissy et al. Captopril and immune regulation
WO2022102887A1 (en) Mass proliferation culture method of nk cells
WO2013100717A2 (en) Composition for preventing and treating multiple sclerosis and autoimmune diseases, containing phellinus igniarius extract
Shen et al. Developmental and communicative interrelations of Ly123 and Ly1 cell sets.
Haynes et al. Mechanisms of corticosteroid action on lymphocyte subpopulations: V. Effects of in Vivo hydrocortisone on the circulatory kinetics and function of naturally occurring and mitogen-induced suppressor cells in man
Golub et al. Systemic administration of human leukocyte interferon to melanoma patients. II. Cellular events associated with changes in natural killer cytotoxicity
WO2020122405A1 (en) Composition for preventing or treating inflammatory diseases, comprising mesenchymal stem cells derived from dedifferentiated stem cells
WO2023277491A1 (en) Composition containing immunosuppressive t cells for preventing or treating immune-related diseases
Nies et al. Impaired immunoglobulin M synthesis by peripheral blood lymphocytes in systemic lupus erythematosus: a primary B-cell defect
Ballas et al. Lymphokine-activated killer (LAK) cells. III. Characterization of LAK precursors and susceptible target cells within the murine thymus.
WO2017074118A1 (en) Pharmaceutical composition containing dendritic cell expressing foxp3 for regulating immunity
EP3500300A1 (en) Immunosuppressive composition for use in treating immunological disorders
Pardo et al. Analysis of in vitro polyclonal B cell differentiation responses to bacterial peptidoglycan and pokeweed mitogen in rheumatoid arthritis
WO2023277490A1 (en) Method for mass production of immunosuppressive t cells
KR20230002080A (en) Composition for preventing or treating Immune Related Disease comprising immune Inhibitory T cells

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 22833547

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 22833547

Country of ref document: EP

Kind code of ref document: A1