Detection of a defect on HLA-E restricted CD8+ T regulatory cells
The present invention provides a composition comprising dendritic cells loaded with hHsp60sp, which dendritic cells are from a subject and have been fixed with paraformaldehyde (PFA). The subject may suffer from an autoimmune disease. Also provided are a method for preparing the composition; recombinant human cells comprising a heterologous gene encoding a fusion protein of HLA-E and hHsp60sp or B7sp, and expressing the fusion protein on the surface of the cells; a method for determining a percentage of maximum inhibition of testing the function of the HLA-E restricted CD8+ Treg cells from a subject, determining whether HLA-E restricted CD8+ Treg cells freshly isolated from a subject are defective, or determining whether defective HLA-E restricted CD8+ Treg cells from a subject are correctable; and a method for correcting defective HLA-E restricted CD8+ Treg cells, treating type 1 diabetes (T1D), or treating multiple sclerosis (MS).
1. A method for determining specific inhibition of specific target cells by HLA-E restricted CD8+ Treg cells freshly isolated from a test subject, comprising:
(a) obtaining a first cell population having an equal number of specific target cells from the cell line having the ATCC accession number PTA-127256 and B721 cells;
(b) culturing the first cell population in the presence of the HLA-E restricted CD8+ Treg cells;
(c) quantifying proliferation of the specific target cells in the first cell population of (b);
(d) obtaining a second cell population having an equal number of control target cells from the cell line having the ATCC accession number PTA-127257 and B721 cells;
(e) culturing the second cell population in the presence of the HLA-E restricted CD8+ Treg cells; and
(f) quantifying proliferation of the control target cells in the second cell population of (e),
wherein the difference between the proliferation of the specific target cells of (c) and the proliferation of the control target cells of (f) indicates the specific inhibition of the specific target cells.
2. The method of claim 1 , wherein the HLA-E restricted CD8+ Treg cells are activated with paraformaldehyde-fixed autologous dendritic cells loaded with hHsp60sp.
3. The method of claim 1 , wherein the test subject suffers from an autoimmune disease.
4. The method of claim 2 , wherein the test subject suffers from an autoimmune disease.
5. The method of claim 4 , wherein the autoimmune disease is selected from the group consisting of type 1 diabetes (T1D), multiple sclerosis (MS), psoriasis, rheumatoid arthritis, lupus, vitiligo, pemphigus and dermatomyositis.
6. The method of claim 2 , wherein the test subject suffers from type 1 diabetes (T1D).
7. The method of claim 2 , wherein the test subject suffers from multiple sclerosis (MS).