CELL SURFACE ANTIGEN OF ACTIVATED IMMUNE CELL AND VARIOUS USES THEREOF

Abstract

The present invention relates to Lrig1 (leucine-rich and immunoglobulin-like domains 1) protein, a cell surface antigen specifically present on the surface of immune cells, and various uses thereof.

Claims

1-35. (canceled)

36. A method for the prevention or treatment of a condition selected from an immune-related disease and a cancer in a subject, the method comprising administering a helper T cell with an Lrig1 protein or a gene that encodes an Lrig1 protein to the subject as an active ingredient.

37. The method according to claim 36, wherein the condition is an immune-related disease.

38. The method according to claim 37, wherein the helper T cell is at least one selected from the group consisting of a Th1 cell, a Th2 cell, a Th17 cell, and a Th22 cell.

39. The method according to claim 37, wherein the immune-related disease is at least one selected from the group consisting of autoimmune disease, graft-versus-host disease, organ transplant rejection, asthma, atopy, and acute or chronic inflammatory disease.

40. The method according to claim 39, wherein the autoimmune disease is at least one selected from the group consisting of rheumatoid arthritis, systemic scleroderma, systemic lupus erythematosus, atopic dermatitis, psoriasis, alopecia areata, asthma, Crohn's disease, Behcet's disease, Sjogren's syndrome, Guillain-Barre syndrome, chronic thyroiditis, multiple sclerosis, multiple myositis, ankylosing spondylitis, fibroblast and nodular multiple arteritis.

41. The method according to claim 36, wherein the condition is a cancer.

42. The method according to claim 41, wherein the helper T cell is at least one selected from the group consisting of a Th1 cell, a Th2 cell, a Th17 cell and a Th22 cell.

43. The method according to claim 41, wherein the cancer is at least one selected from the group consisting of gastric cancer, liver cancer, glioblastoma, ovarian cancer, colorectal cancer, head and neck cancer, bladder cancer, renal cell cancer, breast cancer, metastasis cancer, prostate cancer, pancreatic cancer, melanoma and lung cancer.

44. A diagnostic composition for immune-related diseases or cancer, the composition comprising an agent for measuring the expression levels of the Lrig1 protein on the surface of helper T cell or of the gene that encodes such protein.

45. The diagnostic composition according to claim 44, wherein the diagnostic composition is for immune-related diseases.

46. The diagnostic composition according to claim 45, wherein the helper T cell is at least one selected from the group consisting of a Th1 cell, a Th2 cell, a Th17 cell, and a Th22 cell.

47. A diagnostic kit for immune-related diseases comprising the diagnostic composition according to claim 45.

48. A diagnostic kit for immune-related diseases comprising the diagnostic composition according to claim 46.

49. The diagnostic composition according to claim 44, wherein the diagnostic composition is for cancer.

50. The diagnostic composition according to claim 49, wherein the helper T cell is at least one selected from the group consisting of a Th1 cell, a Th2 cell, a Th17 cell, and a Th22 cell.

51. A diagnostic kit for cancer comprising the diagnostic composition according to claim 49.

52. A diagnostic kit for cancer comprising the diagnostic composition according to claim 50.

53. A method for screening cell therapies for immune-related diseases or for cancer comprising measuring the expression level of Lrig1 protein or the gene encoding it in a separate biological sample.

54. The method of claim 53, wherein the method is for screening cell therapies for immune-related diseases.

55. The method of claim 53, wherein the method is for screening cell therapies for cancer.

Description

DESCRIPTION OF THE DRAWING

[0207] The lone drawing shows the results confirming the inhibitory effect of Th17 cells according to an embodiment of the present invention.

BEST MODE

[0208] The results confirming the inhibitory effect of Th17 cells according to an embodiment of the present invention is shown in the drawing.

MODE FOR INVENTION

[0209] Hereinafter, the present invention will be described in detail by following the embodiments. However, the following embodiments only illustrate the present invention and thus the content of the invention is not restricted nor limited by the following.

EXAMPLES

[Preparation Example] T Cell Subset Cell Culture

[0210] To confirm the effector T cell inhibitory effects of Th17 cells, T cell subsets Th0, Th1, Th2, Th17 and iTreg were prepared. Unlike naturally isolated nTreg, iTreg refers to cells artificially induced to differentiate in a medium containing the following composition.

[0211] T cell subtypes were induced to differentiate by first isolating naïve T cells from the spleen of mice using MACS according to a conventional method. Then the components of Table 1 below were added to a RPM11640 (Invitrogen Gibco, Grand Island, N.Y.) nutritional medium containing 10% Fetal Bovine Serum (FBS; Hyclone, Logan, Utah), and the cells were incubated in a 37° C., 5% CO.sub.2 culture medium for 72 hours to differentiate into individual cells.

TABLE-US-00001 TABLE 1 Differentiated Cell Composition Th0 anti-CD3, anti-CD28 Th1 IL-12, anti-IL-4 antibody Th2 IL-4, anti-IFNβ Th17 IL-6, TGFβ, anti-IFNγ, anti-IL-4 iTreg IL-2, TGFβ

[Example] Confirmation of Effector T Cell Inhibitory Effect

[0212] The differentiated cells from the [Preparation Example] were classified into cells with high expression levels of Lrig1 (Th17 L1+ or iTreg L1+) and cells with low expression levels of Lrig1 (Th17 L1− or iTreg L1−) using the Fluorescence-Activated Cell Sorter; FACS).

[0213] Then, to confirm the effector T cell inhibitory effect, antigen presenting cells (APC) and naïve CD4+T cells were washed twice with PBS to completely remove serum. When the above pellet was dissolved with 1 mL of PBS for CFSE labeling, in the case of 2×10.sup.7 cells, 1 mL of a 5 mM CFSE 0.25 μL mixture was added to 1 mL of PBS and mixed in a 1:1 ratio. However, when the cell count was lower than 2×10.sup.7, it was dissolved with 500 μL of PBS and mixed with 500 μL of the above CPSE+PBS mixture. Then, the mixture was incubated at 37° C. for 10 minutes. In order to inhibit APC division, mitomycin C (0.5 mg/ml) was added in a volume of 100 μL when there were 5×107 cells in 1 mL of PBS and incubated at 37° C. for 20 minutes. After filling 15 mL with pre-chilled medium and storing on ice for 5 minutes, it was washed twice with the culture medium and the cells were counted. Th17L+ cells, Th17L− cells, iTreg L+ cells and iTreg L− cells were added in a ratio of 2:1, 1:1, and 1:2 to CD4+T cells, along with protein and anti-CD3 antibody (0.5 ug/mL). After 3 days, the degree of inhibition of the proliferation of effector T cells was confirmed using CFSE-FL4, and the results are shown in FIG. 1.

[0214] As shown in FIG. 1, compared to the case where the expression level of Lrig1 is low, in Th17 cells with high Lrig1 expression levels, the effector T cell inhibitory effect was exhibited in all cases of 2:1, 1:1 and 1:2 ratios.

[0215] Having described specific parts of the present invention in detail above, it is clear to those skilled in the art that these specific techniques are only preferred embodiments, and that the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

INDUSTRIAL APPLICABILITY

[0216] The immune cells of the present invention have an activated inhibitory function, and the Lrig1 protein of activated immune cells can be used as a new target for immune-related diseases, cancer, or neurodegenerative or neuroinflammatory diseases.