KR20160109358A - A composition for preventing or treating of immune disease - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating immunological disease, containing a tonsil-derived mesenchymal stem cells (T-MSCs) as an active ingredient. The pharmaceutical composition for preventing and treating immunological disease suppresses differentiation and maturation of dendritic cell, inhibits secretion of immune reaction-inducing cytokines secreted by dendritic cell, and weakens functions of dendritic cell-mediated T lymphocytes, thereby regulating an immune reaction. Thus, the pharmaceutical composition shows remarkable effects of preventing and treating immunological disease.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for preventing or treating immune diseases,

The present invention relates to a composition for preventing or treating immunological diseases, which comprises an unilateral derived mesenchymal stem cell as an active ingredient.

Stem cells are the cells that can differentiate into various cells constituting biological tissue and collectively referred to as the undifferentiated cells in the pre-differentiation stage which can be obtained from each tissue of embryo, fetus and adult body.

Recently, transplantation therapy using stem cell and tissue engineering has been developed as a treatment method. However, there is a problem that it is difficult to develop a stable cell therapy agent that does not cause the immune response of the human body because of the excellent therapeutic effect with such transplantation therapy technique. In the meantime, attempts have been made to utilize stem cells as a cell therapy agent, but the confirmation of the ability to regulate immunity is only partially progressed. In particular, there are reports that some of the stem cells are capable of immunosuppression, but there is no report on stem cells having immunomodulating ability yet.

In particular, dendritic cells (DCs) mediated regulation of immune response is an important factor in the prevention and treatment of immune diseases. Dendritic cells (DCs) are the most potent antigen-presenting cells in the immune system. Dendritic cells are immune cells capable of inducing a primary immune response to stimulate naive T cells and inducing immune memory. The ability of dendritic cells to activate a strong immune response is an antigen presenting cell that expresses not only MHC molecules (I / II) but also co-stimulatory molecules at high concentrations on the cell surface , IFN-α, IL-12, IL-18, and other cytokines. These dendritic cells secrete various cytokines such as IFN-α and IL-12, which can induce the generation of antigen-specific cell killer T cells and the proliferation and activation of Th1 cells. Therefore, the regulation of these dendritic cells plays an important role in the regulation of the immune response, and research and development thereof are necessary.

On the other hand, in order to carry out the above-mentioned stem cell research, smooth supply of stem cells is required. However, among mesenchymal stem cells, some stem cells are difficult to use because of their great limitations in obtaining cells for their use. For example, mesenchymal stem cells derived from umbilical cord blood and adipose tissue must be obtained using invasive methods. The most non-invasive cells are mesenchymal stem cells through bone marrow harvesting, but bone marrow harvesting requires anesthesia and causes pain, which limits its use. In order to isolate patient-tailored stem cells, a cell harvesting method using peripheral blood is required as an alternative. However, the number of mesenchymal stem cells that can be isolated from adults is too small, the separation method is not economical, However, since it is difficult to proliferate by an amount that can be used for cell therapy, an alternative method that can increase practicality is needed. In addition, the adult stem cells obtained from the elderly patients have a significantly lower cell potential than the cells obtained from lower ages, and the secretion of various factors and the ability of the stem cells to migrate are lowered. Therefore, the adult stem cells can be separated naturally from the patients of low age or discarded It is necessary to obtain cells from the tissue. In addition, the cells obtained in this manner can be easily quantitated in the experiment, and it is necessary that the differentiation ability is maintained well when the cells are subcultured.

Under such background, it is necessary to research and develop methods for preventing or treating immune diseases by controlling immune responses using specific stem cells.

Korean Patent Publication No. 10-2014-0135263 Korean Patent Publication No. 10-2010-0033471

The present invention provides a pharmaceutical composition for preventing or treating immunological diseases, which comprises an unilateral derived mesenchymal stem cell as an active ingredient.

The present invention also provides a composition for inhibiting dendritic cell differentiation comprising an unilateral derived mesenchymal stem cell as an active ingredient.

The present invention also provides a method for inhibiting dendritic cell differentiation comprising the step of treating a dendritic cell with a composition for inhibiting dendritic cell differentiation comprising an unilateral derived mesenchymal stem cell as an active ingredient.

DISCLOSURE OF THE INVENTION The inventors of the present invention have confirmed the proper types of stem cells capable of preventing or treating immune diseases by controlling the immune response, and confirmed their characteristics. One way stem-derived mesenchymal stem cells inhibit the differentiation and maturation of dendritic cells, Inhibited the secretion of the immune response-induced cytokine, weakened the function of the dendritic cell-mediated T-lymphocyte, and confirmed its availability as a therapeutic agent through inhibition of the immune response, thereby completing the present invention.

In the present invention, a mesenchymal stem cell originating from one side is located in the back of the throat and the nose, and is a tissue that acts primarily as a lymphocyte immune tissue while defending the body from substances such as bacteria that enter from the outside Means an undifferentiated stem cell having the ability to replicate into two or more new cells while having self-replicating ability derived from one-way.

In the present invention, an immunological disease is a disease that becomes a problem when a specific immune response occurs, and is an immune disease that can be caused by the dendritic cell differentiation and activity, and the functional activity of a cell mediated T-lymphocyte. The immunological disease may be any one selected from Type 1 diabetes, rheumatoid arthritis, multiple sclerosis, Crohn's disease, psoriasis.

In the present invention, dendritic cells are antigen-presenting cells having the strongest ability to deliver an antigen among cells existing in the immune system. Dendritic cells are immune cells capable of inducing a primary immune response to stimulate naive T cells and inducing immune memory. Stimulating differentiation into Helper T cells such as T _H 1, T _H 2 and T _H 17 cells of CD4 ⁺ T cells. In addition, MHC molecules (I / II) can be expressed at high concentrations on the cell surface and various cytokines such as IFN-α, IL-12 and IL-18 can be secreted.

Thus, regulation of these dendritic cells plays an important role in regulating the immune response.

In the present invention, CD4 ⁺ T cells are one of the cells involved in the immune response and are largely divided into the first type (T _H 1) and the second type (T _H 2) of our body, Refers to cells involved in the reaction. For example, type 1 cells cause inflammation, enhance phagocytosis and activate cytotoxic T cells, while the type 2 cells mainly cause an antibody reaction, so that when the infectious bacteria are bound by the antibody, So that it can be removed.

The present invention provides a pharmaceutical composition for preventing or treating immunological diseases, which comprises an unilateral derived mesenchymal stem cell as an active ingredient. That is, it can be used as a medicine used for treatment, diagnosis and prevention of cells and tissues produced by isolation, culture and special manipulation from an individual, and can be used for prevention and treatment of immune diseases. Immune diseases in the present invention are as mentioned above. One-way derived mesenchymal stem cells inhibit dendritic cell differentiation and maturation compared to other derived mesenchymal stem cells, inhibit the secretion of immune response-induced cytokines secreted by dendritic cells, and function as dendritic cell mediated T-lymphocytes By modulating the immune response by weakening it, and through it has a remarkable effect as a therapeutic agent for immune diseases. For example, it inhibits the expression of Co-stimulatory molecules and Class II MHC, thereby reducing CD11 ⁺ dendritic cells, inhibiting the differentiation of CD4 ⁺ T cells and inhibiting the secretion of immuno-mediated cytokines.

Since the one-way stem cell of the present invention according to the present invention is easy to acquire the one-way tissue, which is the originating tissue, in comparison with the other stem cells derived from the other, it is possible to recycle the tonsil tissue after the operation and the initial yield is very high. In addition, when a large number of cells are required in the case of inducing sensitization with dendritic cells in vitro, etc., it is possible to use a large amount of cells because of its easy proliferation, and thus, it has a remarkable effect on the prevention and treatment of immune diseases I have.

In the present invention, the one-way-derived mesenchymal stem cells can be extracted from the one side according to methods known in the art.

In the present invention, the prevention means all the actions of inhibiting or delaying the onset of the immune disease upon administration of the composition, and the treatment means all the actions that improve or improve the symptoms of the immune disease upon administration of the composition.

The composition of the present invention may contain 1.0 × 10 ⁵ to 1.0 × 10 ⁹ cells, preferably 1.0 × 10 ⁶ to 1.0 × 10 ⁸ cells per ml.

The pharmaceutical composition for the prevention or treatment of immune diseases may be formulated into a unit dosage form of a pharmaceutical preparation suitable for administration to a patient in accordance with a conventional method in the pharmaceutical field. The preparation may be administered once or several times ≪ / RTI > As formulations suitable for this purpose, injectable preparations, injecting agents and spraying agents are preferred as parenteral administration preparations. In addition, the pharmaceutical composition for preventing or treating the immunological disease may comprise a conventional inert carrier which is pharmaceutically acceptable. The pharmaceutically acceptable carriers and diluents may be biologically and physiologically compatible with the stem cells and the recipient thereof. Diluents include, but are not limited to, saline, aqueous buffer, solvents and / or dispersion media. In addition, preservatives, analgesics, solubilizing agents or stabilizers may be added in the case of injections, and bases, excipients, lubricants or preservatives in the case of topical preparations. The compositions of the present invention can be used without being frozen or frozen for future use. If it is to be frozen, standard cryopreservatives (eg DMSO, glycerol, Epilife ^® cell freezing medium (Cascade Biologics)) may be added to the pre-freezing cell population.

Also, it can be transplanted and administered using a method of administration commonly used in the art, and preferably it can be transplanted or transplanted directly to a diseased part of a patient in need of treatment, but is not limited thereto. For example, the composition according to the present invention can be administered by rectal, subcutaneous, intramuscular, intraperitoneal, intravenous, intraarterial, intracerebral, intramedullary, and the like. In addition, the above administration is possible both in a non-surgical administration using a catheter and in a surgical administration method such as implantation or transplantation after incision of a disease site. The dose may be administered once or several times in the course of 10 ⁷ to 10 ⁹ cells / kg body weight, preferably 10 ⁸ to 10 ⁹ cells / kg body weight. It should be understood, however, that the actual dosage of the active ingredient should be determined in light of various relevant factors such as the disease to be treated, the severity of the disease, the route of administration, the body weight, age and sex of the patient, The scope of the present invention is not limited thereto.

The present invention provides a composition for inhibiting dendritic cell differentiation comprising an unilateral derived mesenchymal stem cell as an active ingredient. The composition for inhibiting dendritic cell differentiation according to the present invention inhibits differentiation into hematopoietic bone marrow cells, preferably dendritic cells derived from monocytes. For example, inhibition of differentiation into immature dendritic cells generated from hematopoietic bone marrow cells by treatment with factors such as interleukin 4 (IL-4) or granulocyte macrophage colony stimulating factor (GM-CSF) and induction from lipopolysaccharide Lt; RTI ID = 0.0 > dendritic < / RTI > The inhibition of differentiation can be inhibited in both intercellular contact conditions and noncontact conditions. The dendritic cell treatment ratio for mesenchymal stem cells can be treated with a ratio of 1: 1 to 1: 100, preferably 1: 5 to 1: 100.

The present invention also provides a method for inhibiting dendritic cell differentiation comprising the step of treating hematopoietic bone marrow cells with a composition for inhibiting dendritic cell differentiation comprising an unilateral derived mesenchymal stem cell as an active ingredient. In the case of the present invention, the composition can be used to inhibit the differentiation of dendritic cells from hematopoietic bone marrow cells, preferably monocytes, and the method of inhibiting differentiation can proceed in vitro.

The pharmaceutical composition for the prevention and treatment of immune diseases of the present invention inhibits the differentiation and maturation of dendritic cells, inhibits the secretion of immune response-induced cytokines secreted by dendritic cells, weakens the function of dendritic cell mediated T-lymphocytes, In the prevention and treatment of immune diseases.

FIG. 1 is an experimental schematic diagram for confirming that one-way-derived mesenchymal stem cells inhibit the differentiation and maturation of dendritic cells under co-culture conditions.
Figure 2 shows CD11b + dendritic cell reduction and CD80, CD86, Class Ⅱ MHC and CD14 expression changes in dendritic cells following treatment with unilateral MSCs under co-culture conditions.
FIG. 3 is an experimental schematic diagram for confirming that the one-way-derived mesenchymal stem cells inhibit the differentiation and maturation of dendritic cells under separate culture conditions.
FIG. 4 is a graph showing the effect of inhibiting the differentiation and maturation of dendritic cells by soluble factor secretion of one-way derived mesenchymal stem cells.
FIG. 5 is an experimental schematic diagram for confirming that the one-way-derived mesenchymal stem cells inhibit dendritic cell mediated T cell differentiation and proliferation.
FIG. 6 shows inhibition of dendritic cell-mediated T cell differentiation and proliferation by addition of one-way derived mesenchymal stem cells and reduction of differentiation into Th1, Th2 and Th17 cells.
FIG. 7 is an experimental schematic diagram for confirming that the one-way-derived mesenchymal stem cells inhibit the secretion of cytokines from mature dendritic cells.
FIG. 8 is a graph showing changes in cytokine expression through cytokine analysis. FIG.
FIG. 9 shows that the expression of GM-CSF, RANTES, interleukin 6 (IL-6) and monocyte chemoattractant protein 1 (MCP-1) is decreased when co-cultured mature dendritic cells and T cells with one-way derived mesenchymal stem cells .
10 shows CD11b + dendritic cell loss and changes in CD80, CD86, Class Ⅱ MHC and CD14 expression in dendritic cells according to the treatment rate of mesenchymal stem cells derived from one side.

Examples and production examples are provided to facilitate understanding of the present invention. The following examples and preparation examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the examples and the production examples.

Example 1 Preparation of Dendritic Cells Derived from Bone Marrow and Mesenchymal Stem Cells

8-week-old female Balb / c mice (OrientBio, Korea) were used for the preparation of bone marrow-derived dendritic cells. All procedures and protocols were performed with approval of the Medical Ethics Committee of ESHWA Women's University (ESM 11-0222). Mice were raised with free food and water under 21-23 ° C and 51-54% humidity and 12 h day / night cycles.

Mice were sacrificed and bone marrow was obtained from the femur and tibia. The thighs and tibia were washed with 5 ml of RPMI 1640 and red blood cells were lysed using ACK buffer (10 ×, 1.5 M NH ₄ Cl, 100 mM KHCO ₃ , 10 mM disodium EDTA). The cell pellet was cultured in 2 × 10 ⁶ cells in a 6-well plate with RPMI 1640 supplemented with 10% FBS, 1% penicillin / streptomycin. Dendritic cells were stained with recombinant mouse GM-CSF (rmGM-CSF, R & D Systems, Minneapolis, Minn., USA) (20 ng / ml) and lipopolysaccharide (LPS) , MO, USA). After another 48 hours, the cells and media were collected for another experiment.

Tonsil-derived mesenchymal stem cells (TMSCs) were isolated from patients who underwent tonsillectomy at post-nasopharyngeal surgery in Ewha Womans University Hospital (4-20-year-old younger group, Passing the examination: ECT 11-53-02). One passage-derived mesenchymal stem cells (4 passages) were preincubated in 6 wells at 5 × 10 ⁵ cells / well for 24 hours under RPMI 1640 supplemented with 10% FBS, 1% penicillin / streptomycin.

<Example 2> Under the co-culture conditions, CD11 ⁺  Identification of Dendritic Cell Differentiation Inhibitory Effect

In Example 1, mouse bone marrow cells (BMC) were cultured with rmGM-CSF (20 ng / ml) for 10 days to induce differentiation from immature dendritic cells (2 x 10 ⁶ cells / well) Lt; / RTI &gt; for 48 hours. One-way derived mesenchymal stem cells (4 × 10 ⁵ cells / well) were added to the culture medium to provide cell-cell contact at day 0 or day 10 of bone marrow cell culture. The experimental procedure is described in Fig.

Flow cytometry was performed to confirm the phenotype.

Cells were cultured with 2 ug / ml of the following Fluorochrome-conjugated antibodies: FITC-anti-mouse CD11b (M1 / 70, Rat IgG _2b , eBiosciences, San Diego, CA) _{1, BD Biosciences, Franklin Lakes,} NJ), APC-anti-mouse CD11c (N418, Hamster IgG, BioLegend, San Diego, CA), PerCP-anti-mouse CD80 (16-1 OA1, Hamster IgG 2, BD Biosciences) , PE-anti-mouse CD86 ( GL1, Rat IgG 2a, BD Biosciences), PE-anti-mouse CD14 (rmC5-3, Rat IgG 1, BD Biosciences), PE-anti-mouse class II major histocompatibility complex (MHC) _{(2G9, Rat IgG 2a, BD} Biosciences), PE-anti-mouse CCR7 (4B12, Rat IgG 2a, R & D Systems). Unstained cells were used in the control group. Cells were centrifuged at 300 xg for 5 min at room temperature, washed and fixed with 1% paraformaldehyde under PBS.

For intracellular staining, the cells were fixed with 4% paraformaldehyde in PBS for 20 min on ice and incubated with 20% permeabilization buffer (0.5% saponin, 1% bovine serum albumin [BSA] in PBS) . Cells were incubated with Fluorochrome-conjugated antibody for 30 min under ice: FITC-anti-mouse CD4 (RM4.5, Rat IgG _2a , BD Biosciences), APC-anti-mouse interferon gamma (IFN- , Rat IgG ₁ , BioLegend), PE-anti-mouse interleukin 4 (IL-4) (11B11, Rat IgG ₁ , BD Biosciences) Rat IgG ₁ , BioLegend). After staining, cells were washed with 0.5% FBS in PBS and analyzed with a flow cytometer with Cell Quest software (BD Biosciences).

The results are shown in Fig.

As can be seen in Fig. 2A, the proportion of CD11b ⁺ bone marrow cells significantly increased from 38.9 + 4.8% to 90.7 + 1.9% when GM-CSF (20 ng / ml) was added.

However, the addition of T-MSCs at day 0 inhibited the proliferation of CD11b ⁺ cells (55.1 ± 8.3%), which was similar in the addition of LPS (57.7 ± 8.9% ). In other words, it was confirmed that the mesenchymal stem cell derived from one end of the cell suppresses the dendritic cell maturation by GM-CSF-induced dendritic cell-derived dendritic cell differentiation and LPS stimulation under cell-cell contact condition.

CD80, CD86, Class Ⅱ MHC, and CD14 expression in the rat brain. LPS stimulation increased the expression of CD80 (56.8 ± 5.2%), CD86 (69.7 ± 3.4%) and class II MHC (64.5 ± 17.31%) in dendritic cells. In the case of CD14 ⁺ cells, the ratio was unchanged, indicating that dendritic cells were not stimulated by LPS.

On the other hand, addition of one - way mesenchymal stem cells from day 0 significantly reduced the expression of class II MHC while inhibiting the expression of LPS - induced maturation - induced CD80 and CD86. In other words, it shows that the one-way derived mesenchymal stem cells play a pivotal role in suppressing the differentiation of dendritic cells.

In other words, it was confirmed that the mesenchymal stem cells derived from uniloculation inhibited the maturation of the bone marrow-derived dendritic cells from the suppression of the expression of the co-expressed factors and class II MHC under cell contact conditions.

<Example 3> Preparation of CD11 ⁺  Identification of Dendritic Cell Differentiation Inhibitory Effect

It was also confirmed by the soluble factors and the like that the mesenchymal stem cells derived from unilamellar can inhibit the differentiation of dendritic cells from bone marrow cells under separate culturing conditions.

A transwell plate (Corning, Acton, MA) with a 0.4 um pore size polycarbonate membrane was used to prevent contact between one-way derived mesenchymal stem cells and bone marrow derived dendritic cells. Bone marrow-derived dendritic cells and one-way derived mesenchymal stem cells were placed at the top and bottom of the chamber at a ratio of 5: 1, respectively, and cultured under RPMI1640 supplemented with 10% FBS, 1% penicillin / streptomycin. LPS (1 μl / ml) was added to the upper chamber for 10 days to induce maturation of bone marrow-derived dendritic cells. The experimental conditions are shown in Fig.

Flow cytometry was performed as in Example 2 above. The results are shown in Fig. Derived mesenchymal stem cells inhibited the proliferation of CD11b ⁺ cells in bone marrow-derived dendritic cells induced by GM-CSF. Similar to cell contact conditions, dendritic cell differentiation and maturation by GM-CSF and LPS stimulation . &Lt; / RTI &gt;

Example 4 CD4 < RTI ID = 0.0 > ⁺  Identification of T cell differentiation and dendritic cell mediated T cell differentiation inhibitory effect

T cells were isolated from mouse spleen and labeled with CFSE for cell division in order to confirm the effect of one-way derived mesenchymal stem cells co-cultured with bone marrow-derived dendritic cells in T cell proliferation.

For isolation of T cells, the spleen was obtained from 8 week old female Balb / c mice and ground in 5 ml RPMI1640 medium. The cell suspension was washed and resuspended in 600 ul MACS sorting buffer (0.5% BSA in PBS, 2 mM EDTA, pH 7.2), followed by addition of anti-CD4 or anti-CD3 microbead solution (10 ul per 10 ⁷ cells). Cells were treated with ice for 15 minutes, washed and resuspended in 1 ml sorting buffer. Cells were separated according to the manufacturer's manual using a MACS magnetic column (Miltenyi Biotec, Auburn, CA, USA). Isolated cells were stained with PE-conjugated anti-mouse CD4 antibody (GK 1.5, Rat IgG _2b , BioLegend) and analyzed by flow cytometry to determine cell population purity.

To measure proliferation, 2 × 10 ⁸ cells were stained with 5 μM carboxyfluorescein succinimidyl ester (CFSE, Invitrogen, Carlsbad, Calif., USA) for 5 min at room temperature and 1 ml FBS was added to stop the reaction. Cells are washed three times with RPMI1640 containing 10% FBS and were transferred to 6 well culture plates ^{(2 × 10 6 cell / well} ). One-way derived mesenchymal stem cells or bone marrow derived dendritic cells were added and cultured for 48 hours. For T cell stimulation, 5 μg / ml LEAF ^™ purified anti-mouse CD3 (17A2, Rat IgG _2b , BioLegend) and LEAF ^™ purified anti-mouse CD28 (37.51, Syrian Hamster IgG, BioLegend) Was diluted in RPMI 1640 containing antibiotics and added to the cells. The number of cells was 2 × 10 ⁶ cell / well with respect to ^{4 × 10 5 cell / well,} 2 with respect to the bone marrow-derived dendritic cells × 10 ⁶ cell / well and T-cell with respect to the one-way-derived MSCs. After 48 hours, cells were harvested for flow cytometry and supernatants were collected for Cytokine assay. The experimental method is shown in FIG.

The results of the analysis are shown in FIG.

Figure 6A shows flow cytometry results. T cell proliferation, which is indicated by a decrease in CFSE intensity, decreased from 22.99% to 5.3% after the addition of the unilateral MSCs to the culture medium. In other words, 22.99% CD4 + T cells were cultured with bone marrow-derived dendritic cells and stimulated with anti-CD3 / anti-CD28, whereas at day 0, cell division was reduced to 5.3% in co-culture with one-way mesenchymal stem cells.

CD4 ⁺ T cells were isolated from mouse spleen and co-cultured with bone marrow-derived dendritic cells to evaluate the differentiation potential of bone marrow-derived dendritic cells and CD4 ⁺ T cells, and anti-CD3 / anti- The results of stimulation under the presence or absence of stem cells are shown in Fig. 6B.

As a result of flow cytometry analysis, the percentage of CD4 ⁺ INF-γ ⁺ (T _H 1 cell), CD4 ⁺ IL-4 ⁺ (T _H 2 cell), and CD4 ⁺ IL17A ⁺ (T _H 17) Derived dendritic cells and after stimulation with anti-CD3 / anti-CD28 antibody. However, the proportion of differentiated T cells decreased when co-cultured with bone marrow-derived dendritic cells and pre-treated with one-way derived mesenchymal stem cells.

From the above results, it was confirmed that CD4 ⁺ T cell differentiation and dendritic cell mediated T cell differentiation inhibitory effect by one-way-derived mesenchymal stem cells were inhibited.

Example 5 Confirmation of Cytokine Secretion Modulation from Matured Dendritic Cells by Mesenchymal-Derived Mesenchymal Stem Cells

The culture medium and the human Cytokine array C1 kit (RayBiotech, Norcross, GA, USA) collected in the above experiment were kept at room temperature, and the assay was performed according to the manufacturer's manual. The developed membranes were analyzed using a chemiluminescence imaging system (LAS-3000, Fujifilm, Japan). The experimental method is as shown in Fig.

Results of bone marrow-derived dendritic cell secretory analysis on T cell stimulation are shown in FIG.

IL-6, MCP-1 (CCL2), monocyte chemoattractant protein 2 (MCP-1), GM-CSF, growth-regulated oncogene α (GRO-α, CXCL1), RANTES (CCL5), interleukin 8 2, CCL8), and monocyte chemoattractant protein 3 (MCP-3, CCL7) were increased by T cell stimulation.

The result of quantitating the change of the cytokine is shown in Fig. Among cytokines, GM-CSF, IL-6, RANTES and MCP-1 were significantly decreased by the addition of unilateral MSCs. In other words, the effect of inhibiting the secretion of immuno - mediated cytokine by dendritic cells induced by T - cell stimulation was confirmed in one - way derived mesenchymal stem cells.

<Example 6> Confirmation of CD11b + dendritic cell reduction according to the treatment ratio of mesenchymal stem cells

The expression levels of CD11b + dendritic cells and CD80, CD86, Class Ⅱ MHC and CD14 were examined by varying the dendritic cell treatment ratio of one-way derived mesenchymal stem cells from 1: 1 to 1: 100 in the same manner as in Example 1 above The results are shown in Fig.

As shown in FIG. 10, in the ratio of 1: 1 to 1: 100, the dendritic cell-derived mesenchymal stem cell treatment showed remarkable dendritic cell reduction, and in particular, the ratio was between 1: 5 and 1: 100 It was excellent.

Claims (8)

A pharmaceutical composition for preventing or treating immunological diseases, which comprises mesenchymal stem cell derived from one side as an active ingredient. The pharmaceutical composition according to claim 1, wherein the immune disease is an immune disease caused by promotion of dendritic cell activity. The pharmaceutical composition according to claim 1 or 2, wherein the immune disease is any one selected from type 1 diabetes, rheumatoid arthritis, multiple sclerosis, Crohn's disease or psoriasis. The pharmaceutical composition according to claim 3, wherein the immune disease prevention or treatment is by inhibiting dendritic cell differentiation. A composition for inhibiting dendritic cell differentiation comprising an unilateral derived mesenchymal stem cell as an active ingredient. 6. The composition for inhibiting dendritic cell differentiation according to claim 5, wherein said dendritic cell differentiation inhibition is inhibition of dendritic cell differentiation from hematopoietic bone marrow cells. A method for inhibiting dendritic cell differentiation comprising the step of treating hematopoietic bone marrow cells with a composition for inhibiting dendritic cell differentiation comprising an unilateral derived mesenchymal stem cell as an active ingredient. [8] The method according to claim 7, wherein the hematopoietic myeloma cells are treated with the mesenchymal stem cells from the one side to the dendritic cells at a ratio of 1: 1 to 1: 100.

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