Methods of preparing anti-human papillomavirus antigen t cells

Abstract

Disclosed are methods of preparing an isolated population of human papillomavirus (HPV)-specific T cells comprise dividing an HPV-positive tumor sample into multiple fragments; separately culturing the multiple fragments; obtaining T cells from the cultured multiple fragments; testing the T cells for specific autologous HPV-positive tumor recognition; selecting the T cells that exhibit specific autologous HPV-positive tumor recognition; and expanding the number of selected T cells to produce a population of HPV-specific T cells for adoptive cell therapy. Related methods of treating or preventing cancer using the T cells are also disclosed.

1 . A method of preparing a population of human papillomavirus (HPV)-specific T cells, the method comprising:
 dividing an HPV-positive tumor sample into multiple fragments;   separately culturing the multiple fragments in the presence of only one cytokine;   obtaining T cells from the cultured multiple fragments;   testing the T cells for one or both of specific autologous HPV-positive tumor recognition and HPV antigen recognition;   selecting the T cells that exhibit one or both of specific autologous HPV-positive tumor recognition and HPV antigen recognition; and   expanding the number of selected T cells to produce a population of HPV-specific T cells.   
     
     
         2 . The method according to  claim 1 , wherein expanding the number of selected T cells comprises expanding the number of cells using one or both of (i) irradiated allogeneic feeder cells and (ii) irradiated autologous feeder cells and one or both of (iii) OKT3 antibody and (iv) interleukin (IL)-2. 
     
     
         3 . The method according to  claim 1 , wherein the cytokine is IL-2. 
     
     
         4 . A method of preparing a population of HPV-specific T cells, the method comprising:
 dividing an HPV-positive tumor sample into multiple fragments;   separately culturing the multiple fragments;   obtaining T cells from the cultured multiple fragments;   testing the T cells for one or both of specific autologous HPV-positive tumor recognition and HPV antigen recognition;   selecting the T cells that exhibit one or both of specific autologous HPV-positive tumor recognition and HPV antigen recognition; and   expanding the number of selected T cells using one or both of (i) OKT3 antibody and (ii) interleukin (IL)-2 to produce a population of HPV-specific T cells.   
     
     
         5 . The method according to  claim 4 , wherein separately culturing the multiple fragments comprises culturing the multiple fragments in the presence of only one cytokine. 
     
     
         6 . The method according to  claim 5 , wherein the cytokine is IL-2. 
     
     
         7 . The method according to  claim 4 , wherein expanding the number of selected T cells further comprises using one or both of irradiated allogeneic feeder cells and irradiated autologous feeder cells. 
     
     
         8 . The method according to  claim 1 , wherein the population of HPV-specific T cells recognize HPV 16-positive cancer cells. 
     
     
         9 . The method according to  claim 1 , wherein the population of HPV-specific T cells recognize HPV 16 E6. 
     
     
         10 . The method according to  claim 1 , wherein the population of HPV-specific T cells recognize HPV 16 E7. 
     
     
         11 . The method according to  claim 1 , wherein the population of HPV-specific T cells recognize HPV 18-positive cancer cells. 
     
     
         12 . The method according to  claim 1 , wherein the population of HPV-specific T cells recognize HPV 18 E6. 
     
     
         13 . The method according to  claim 1 , wherein the population of HPV-specific T cells recognize HPV 18 E7. 
     
     
         14 .- 19 . (canceled) 
     
     
         20 . A method of treating or preventing cancer in a mammal, the method comprising preparing a population of HPV-specific T cells according to the method of  claim 1  and administering the population of T cells to the mammal in an amount effective to treat or prevent cancer in the mammal. 
     
     
         21 . A method of treating or preventing cancer in a mammal, the method comprising:
 dividing an HPV-positive tumor sample into multiple fragments;   separately culturing the multiple fragments;   obtaining T cells from the cultured multiple fragments;   testing the T cells for one or both of specific autologous HPV-positive tumor recognition and HPV antigen recognition;   selecting the T cells that exhibit one or both of specific autologous HPV-positive tumor recognition and HPV antigen recognition;   expanding the number of selected T cells to produce a population of HPV-specific T cells for adoptive cell therapy; and   administering the expanded number of T cells to the mammal in an amount effective to treat or prevent cancer in the mammal.   
     
     
         22 . The method according to  claim 20 , wherein the cancer is cancer of the uterine cervix, oropharynx, anus, anal canal, anorectum, vagina, vulva, or penis. 
     
     
         23 . The method according to  claim 20 , wherein the cancer is an HPV-positive cancer. 
     
     
         24 . The method according to  claim 20 , further comprising administering to the mammal nonmyeloablative lymphodepleting chemotherapy prior to administering the T cells. 
     
     
         25 . The method according to  claim 20 , wherein either
 (a) the T cells are modified to express a T cell growth factor that promotes the growth and activation of the T cells; or   (b) a T cell growth factor that promotes the growth and activation of the T cells is administered to the mammal either concomitantly with the T cells or subsequently to the T cells.