Methods for selective expansion of γδ T-cell populations and compositions thereof
The present invention relates to methods for the selective expansion of γδ T-cell population(s), compositions and admixtures thereof and methods for using the same as a therapeutic. Non-engineered and engineered, enriched γδ T-cell populations of the disclosure are useful in the treatment of various cancers, infectious diseases, and immune disorders.
1. An ex vivo method for producing an enriched δ1 γδ T-cell population from an isolated mixed cell population without prior selection of γδ T-cells and/or depletion of non-γδ T-cells, and under culture conditions that do not comprise IL-4 and wherein said cell population has not been exposed to exogenous IL-4 prior to expansion, said method comprising directly contacting the mixed cell population with at least one antibody that selectively expands δ1 T-cells without significant expansion of δ2 T-cells by binding to an epitope specific to a δ1 variable region of a δ1 TCR to provide clinically-relevant levels of an enriched δ1 γδ T cell population, wherein the at least one antibody comprises a heavy chain variable region/light chain variable region (HCVR/LCVR) sequence pair selected from the group consisting of SEQ ID NOs: 153/180, 154/181, 155/182, 156/183, 157/184, 158/185, 159/186, 160/187, 161/188, 162/189, 163/190, 164/191, 165/192, 166/193, 167/194, 168/195, 169/196, 170/197, 171/198, 172/199, 173/200, 174/201, 175/202, 176/203, 177/204, 178/205, and 179/206.
2. The method of claim 1 , wherein the percentage of δ1 T cells in the enriched γδ T cell population is greater than 60% δ1 T-cells of total viable cells.
3. The method of any one of the preceding claims, wherein said method does not comprise expanding the enriched δ1 γδ T-cell population by an antigen presenting cell or an aminophosphate.
4. The method of claim 1 , wherein said at least one antibody that selectively expands δ1 T-cells is immobilized on a surface.
5. The method of claim 1 , wherein said isolated mixed cell population is selected from a peripheral blood sample, a cord blood sample or a tumor.
6. The method of claim 1 , wherein said enriched δ1 γδ T-cell population comprise polyclonal TCR diversity.
7. The method of claim 1 , wherein the enriched δ1 γδ T-cell population is further formulated for administration to a subject.
8. The method of claim 1 , wherein the enriched δ1 γδ T-cell population includes a therapeutically effective amount of γδ T-cells.
9. The method of claim 1 , wherein said δ1 γδ T-cell population is further engineered to stably express one or more tumor recognition moieties.
10. The method of claim 1 , further comprising:
directly contacting at least a portion of the enriched δ1 γδ T-cell population with antigen presenting cells (APCs) in a second γδ T-cell expansion, thereby producing a second enriched γδ T-cell population,
wherein the second enriched γδ T-cell population comprises a clinically relevant number of γδ T-cells.
11. The method of claim 1 , further comprising:
directly contacting at least a portion of the enriched δ1 γδ T-cell population with one or more agents that (a) expand γδ T-cells, and/or (b) deplete αβ T-cells.
12. The method of claim 1 , wherein an antibody that selectively expands δ1 T-cells is an antibody that specifically recognizes the δ1 chain variable region and i) the CDR3 loop created by the restricted juxtaposition of V(D)J1 segments, or ii) the CDR3 loop created by the juxtaposition of V(D)J1 and V(D)J2 but not V(D)J3 segments, or iii) the CDR3 loop created by the juxtaposition of V(D)J1, V(D)J2 and V(D)J3 segments.
13. The method of claim 1 , wherein the antibody binds to an epitope comprising residues Arg71, Asp72 and Lys120 of the δ1 variable region.
14. The method of claim 1 , wherein the antibody has reduced binding to a mutant 61 TCR polypeptide comprising a mutation at K120 at the delta J1 and delta J2.
15. The method of claim 1 , wherein the epitope is selected from a δ1 TCR Bin 1 δ1 epitope, Bin 1b δ1 epitope, Bin 2 δ1 epitope, Bin 2b δ1 epitope, Bin 2c δ1 epitope, Bin 3 δ1 epitope, Bin 4 δ1 epitope, Bin 5 δ1 epitope, Bin 6 δ1 epitope, Bin 7 δ1 epitope, Bin 8 δ1 epitope, or Bin 9 δ1 epitope of a human δ1 TCR.