Polyclonal gamma delta t cells for immunotherapy

Abstract

Provided herein is a method of expanding clinically-relevant quantities of polyclonal γδ T cells that have anti-tumor, anti-viral, and anti-bacterial reactivity. Polyclonal γδ T cells can target a variety of tumors, including solid tumors as well as other conditions, such as viral and bacterial infections.

1 . A cell composition comprising at least about 10 6  purified γδ T cells, wherein the γδ T cells comprise Vδ populations selected from Vδ1, Vδ2, and Vδ1 neg Vδ2 neg  TCR subsets. 
     
     
         2 . The cell composition of  claim 1 , wherein the composition does not contain NK cells or αβ T cells. 
     
     
         3 . The cell composition of  claim 1 , wherein the γδ T cells express CD3. 
     
     
         4 . The cell composition of  claim 1 , wherein the γδ T cells do not express CD57 or PD-1. 
     
     
         5 . A pharmaceutical composition comprising the cell composition of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         6 . A method of expanding a γδ T cells to sufficient numbers for use in therapy comprising:
 (a) obtaining a sample of cells comprising a first polyclonal γδ T-cell population; 
 (b) isolating a first polyclonal γδ T-cell population; and 
 (c) culturing the first polyclonal γδ T-cell population with artificial antigen presenting cells (aAPCs) in the presence of interleukin-2 (IL-2) and interleukin-21 (IL-21), thereby inducing proliferation of the γδ T cells to sufficient numbers for use in therapy. 
 
     
     
         7 . The method of  claim 6 , further comprising depleting the sample of cells of CD56- and TCRα-expressing cells. 
     
     
         8 . The method of  claim 6 , wherein the sample of cells is a peripheral blood sample, an umbilical cord blood sample, or a tissue sample. 
     
     
         9 . The method of  claim 6 , wherein the sample of cells is obtained from a single subject. 
     
     
         10 . The method of  claim 6 , wherein the first polyclonal γδ T-cell population comprises about 10 4  to about 10 6  γδ T cells. 
     
     
         11 . The method of  claim 6 , wherein the aAPCs are transgenic K562 cells. 
     
     
         12 . The method of  claim 6 , wherein the aAPCs express CD137L. 
     
     
         13 . The method of  claim 12 , wherein the aAPCs further express CD 19, CD64, CD86,-and mIL15. 
     
     
         14 . The method of  claim 11 , wherein the aAPCs express at least one anti-CD3 antibody clone. 
     
     
         15 . The method of  claim 14 , wherein the at least one anti-CD3 antibody clone is OKT3 and/or UCHT1. 
     
     
         16 . The method of  claim 6 , wherein culturing the first polyclonal γδ T-cell population with aAPCs comprises culturing the cells at a ratio of about 10:1 to about 1:10 (γδ T cells to aAPCs). 
     
     
         17 . The method of  claim 6 , wherein culturing the first polyclonal γδ T-cell population with aAPCs results in at least a 100-fold increase in the number of polyclonal γδ T cells. 
     
     
         18 . A method of treating a disease in a patient comprising: administering an effective amount of a cell composition produced by the method of  claim 6  to a patient in need thereof. 
     
     
         19 . The method of  claim 18 , wherein the disease is cancer. 
     
     
         20 . The composition of  claim 19 , wherein the cancer is ALL, B-ALL, CML, colon cancer, ovarian cancer, or pancreatic cancer.