Compositions and methods for diminishing an immune response
The invention is based upon the discovery that T regulatory type 1 (Tr1) cells express particular cell surface markers that allow for their selection, enrichment, isolation, purification and administration. The ability to use the particular markers described herein to select, enrich, isolate, purify and administer Tr1 cells allows for improved methods of Tr1 therapies for treating a wide variety of diseases and disorders.
1. A method of isolating an enriched population of human Tr1 cells from a biological sample of a subject by flow cytometry, comprising the steps of:
a. obtaining a T cell-containing biological sample of a subject,
b. gating on CD4+CD45RA- cells and,
c. isolating CD49b+ and LAG-3+ cells among the gated CD4+ and CD45RA- cells.
2. The method of claim 1 , comprising the additional step of isolating cells from the biological sample of the subject that express the cell surface marker CD226.
3. The method of claim 2 , wherein the cells express the cell surface marker CD226 at a level greater than the level of CD226 expressed by a comparator cell population.
4. The method of claim 3 , wherein the comparator cell population is at least one selected from the group consisting of CD49b − LAG-3 − T cells and T H 0 cells.
5. The method of claim 1 , wherein greater than 90% of the cells in the enriched population of Tr1 cells express the cell surface markers CD4, and CD49b, and LAG-3.
6. The method of claim 1 , wherein greater than 95% of the cells in the enriched population of Tr1 cells express the cell surface markers CD4, and CD49b, and LAG-3.
7. The method of claim 1 , wherein greater than 98% of the cells in the enriched population of Tr1 cells express the cell surface markers CD4, and CD49b, and LAG-3.
8. The method of claim 1 , wherein greater than 99% of the cells in the enriched population of Tr1 cells express the cell surface markers CD4, and CD49b, and LAG-3.
9. The method of claim 1 , wherein isolating cells from the biological sample of the subject employs the use of an antibody that specifically binds to a cell surface marker.
10. The method of claim 9 , wherein the cell surface marker is at least one selected from the group consisting of CD4, CD49b, and LAG-3.
11. The method of claim 1 , wherein isolating cells from the biological sample of the subject employs the use of fluorescence-activated cell sorting (FACS).
12. The method of claim 1 , wherein the biological sample is at least one selected from the group consisting of blood, bone marrow, cord blood, lymph nodes, thymus, and spleen.
13. The method of claim 1 , wherein the T cell-containing biological sample is a cell preparation consisting of antigen-specific IL-10-anergized T cells generated with recombinant IL-10-treated dendritic cells (said cell preparation is also named DC-10).