Engineered γδ T-cells
The present invention relates to engineered γδ T-cell(s) and methods for using the same as a therapeutic with a potent and selective ability to target an antigen of choice. Engineered γδ T-cells of the disclosure are useful in the treatment of various cancers, infectious diseases, and immune disorders. Also disclosed are methods for expanding engineered and non-engineered γδ T-cell(s) populations to therapeutically useful quantities. An engineered γδ T-cell of the disclosure can be a universal donor, and can be administered to a subject with any MHC haplotype.
1. An ex vivo method for producing an expanded δ1 γδ T-cell population, comprising
a) providing a sample of a human subject, wherein the sample comprises γδ T-cells,
b) selectively expanding
the δ1 γδ T-cells by culturing cells of the sample in the presence of a cytokine selected from IL-2, IL-7, IL-9, IL-12, IL-15, IL-18, IL-21, and IL-33, and at least one antibody which selectively expands the δ1 γδ T-cells to produce a culture comprising the expanded δ1 γδ T-cell population;
wherein the culture conditions do not comprise IL-4 and said cells have not been exposed to exogenous IL-4 prior to expansion, and wherein:
said at least one antibody which selectively expands δ1 γδ T-cells is selected from TS-1, TS8.2 or R9.12 antibodies which bind to a specific epitope of a δ1 TCR chain, to selectively expand δ1 T-cells; and
wherein the culture comprising the expanded γδ T-cell population comprises a percentage of δ1 γδ T-cells, wherein the percentage of δ1 γδ T-cells is greater than 60% of total viable cells of the culture, optionally wherein the percentage of δ1 γδ T-cells is greater than 80% or 90%, optionally, of total viable cells of the culture.
2. The method of claim 1 , wherein said method does not comprise expanding the γδ T-cell population by an antigen presenting cell, an aminophosphate, or a nitrogen containing bisphosphonate.
3. The method of claim 1 , wherein said at least one antibody which selectively expands δ1 γδ T-cells are immobilized on a surface.
4. The method of claim 1 , wherein said sample is selected from a peripheral blood sample, a cord blood sample or a tumor.
5. The method of claim 1 , wherein the expanded γδ T-cell population is further formulated for administration to a subject.
6. The method of claim 1 , wherein the expanded γδ T-cell population includes a therapeutically effective amount of γδ T-cells.
7. The method of claim 6 , wherein said γδ T-cell population is engineered to stably express one or more tumor recognition moieties.
8. The method of claim 7 , wherein the expanded γδ T-cell population comprises at least about 1×10 7 γδ T-cells.
9. The method of claim 1 , wherein said cytokine is selected from the group consisting of IL-2, IL-7, IL-15, and IL-21.
10. The method of claim 9 , wherein said cytokine is selected from the group consisting of IL-2, IL-7, and IL-15.
11. The method of claim 1 , further comprising:
enriching the γδ T-cells prior to selectively expanding the δ1 γδ T-cells.
12. The method of claim 11 , wherein the enriching includes removing one or more of red blood cells, NK cells, αβ cells, B cells, monocytes, and macrophages.