Identifcation of CD8
This invention provides, among other things, methods for the identification and isolation of viable putative long-lived antigen-specific memory CD8 + T cell subsets (CMhi and EMhi) with high surface expression of CD161 and/or IL-18Rα and the capacity to rapidly efflux the fluorescent dye Rh123.
1. A composition comprising: a) a physiological acceptable carrier and b) an isolated population of long-lived memory CD8+ CD161 hi IL-18Rα hi T cells, wherein:
the long-lived memory CD8+ CD 161 hi IL-18Rα hi T cells make up at least 30% of the total CD8+ T cells in the composition;
the long-lived memory CD8+ CD161 hi IL-18Rα hi T cells comprise CD95 hi memory cells, the CD95 hi memory cells being capable of proliferating in response to IL-7 or IL-15 and comprising a population of cells comprising an engineered immunoreceptor; and where the isolated population of long-lived memory CD8+ CD161 hi IL-18Rα hi T cells comprises high CD28 surface expression
and high MDR-1 mRNA levels, as compared to a CD8+ T cell population with low surface expression of IL-18Rα.
2. The composition of claim 1 , wherein the isolated long-lived memory CD8+ CD161 hi IL-18Rα hi T cells are CD127 + , CD25 neg , bcl 2 hi , perforin neg/low , granzyme A int , granzyme B int/neg and NKG2D int .
3. The composition of claim 1 , wherein the isolated long-lived memory CD8+ CD161 hi IL-18Rα hi T cell population has increased expression of CD43, CD44, CD46, CD148, and CD162 as compared to a CD8+ T cell population with low surface expression of IL-18Rα.
4. The composition of claim 1 , wherein the isolated long-lived memory CD8+ CD 161 hi IL-18Rα hi T cell population lacks expression of CD57, CD103, and CD69 as compared to a CD8+ T cell population with low surface expression of IL-18Rα.
5. The composition of claim 1 , wherein the isolated long-lived memory CD8+ CD161 hi IL-18Rα hi T cell population has increased expression of CD122 as compared to a CD8+ T cell population with low surface expression of IL-18Rα.
6. The composition of claim 1 , wherein the CD95 hi memory cells comprise CD62L + , CD45RA int/neg , CD45RO int/hi central memory cells.
7. The composition of claim 1 , wherein said CD95 hi memory cells comprise CD 62L − , CD45RA int/neg , CD45RO int/hi effector memory cells.
8. The composition of claim 6 , wherein said CD95 hi memory cells comprise CD 62L − , CD45RA int/neg , CD45RO int/hi effector memory cells.
9. The composition of claim 1 , wherein the engineered immunoreceptor is specific for a tumor associated antigen.
10. The composition of claim 1 , wherein the engineered immunoreceptor is an antigen specific T-cell receptor.
11. The composition of claim 1 , wherein the isolated long-lived memory CD8+ CD 161 hi IL-18Rα hi T cells are at least 40% of the total CD8+ T cells in the composition.
12. The composition of claim 1 , wherein the isolated long-lived memory CD8+ CD 161 hi IL-18Rα hi T cells are at least 50% of the total CD8+ T cells in the composition.
13. The composition of claim 1 , wherein the isolated long-lived memory CD8+ CD 161 hi IL-18Rα hi T cells are at least 80% of the total CD8+ T cells in the composition.
14. The composition of claim 1 , wherein the isolated long-lived memory CD8+ CD 161 hi IL-18Rα hi T cells have enhanced proliferation in response to a cytokine selected from the group consisting of IL-12, IL-18, IL-23, and combinations thereof, as compared to a CD8+ T cell population with low surface expression of IL-18Rα.