Preparation of a therapeutic composition for treating autoimmune diseases

A method for preparing an autologous dendritic cell/hHsp60sp complex for correction of defective HLA-E restricted CD8+ Treg cells, comprising:

• (a) preparing autologous immature dendritic cells from a test human individual having defective HLA-E restricted CD8+ Treg cells, wherein the prepared autologous immature dendritic cells are loaded with human heat shock protein 60 signal peptide (hHsp60sp) and fixed with paraformaldehyde (PFA) to form an autologous dendritic cell/hHsp60sp complex;
• (b) freshly isolating defective HLA-E restricted CD8+ Treg cells from the test human individual (“test HLA-E restricted CD8+ Treg cells”);
• (c) testing the prepared autologous dendritic cell/hHsp60sp complex of step (a) for correcting the test HLA-E restricted CD8+ Treg cells in a CD8+ T cell specificity assay, wherein the CD8+ T cell specificity assay comprises specific target cells from the cell line having the ATCC accession number PTA-127256 and control target cells from the cell line having the ATCC accession number PTA-127257, wherein the CD8+ T cell specificity assay comprises:
• (i) activating the test HLA-E restricted CD8+ Treg cells with the prepared autologous dendritic cell/hHsp60sp complex, whereby activated test HLA-E restricted CD8+ Treg cells are produced;
• (ii) determining the percentage of maximum inhibition for the specific target cells and the activated test HLA-E restricted CD8+ Treg cells,
• (iii) activating the test HLA-E restricted CD8+ Treg cells with fixed autologous immature dendritic cells loaded with B7sp, whereby activated control HLA-E restricted CD8+ Treg cells are produced;
• (iv) determining the percentage of maximum inhibition for the control target cells and the activated control HLA-E restricted CD8+ Treg cells, and
• (v) determining a normalized percentage of the maximum inhibition for the test HLA-E restricted CD8+ Treg cells by subtracting the percentage of maximum inhibition for the control target cells and the activated control HLA-E restricted CD8+ Treg cells from the percentage of maximum inhibition for the specific target cells and the activated test HLA-E restricted CD8+ Treg cells, wherein a normalized percentage of maximum inhibition for the test HLA-E restricted CD8+ Treg cells greater than 50% of that for the control HLA-E restricted CD8+ Treg cells indicates that the prepared autologous immature dendritic cells correct the test HLA-E restricted CD8+ Treg cells;
• and
• (d) obtaining the autologous dendritic cell/hHsp60sp complex that corrects the test HLA-E restricted CD8+ Treg cells in step (c).