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 - 19 . (canceled) 
     
     
         20 . A method of treating or preventing a head and neck cancer in a mammal, the method comprising:
 (a) dividing the head and neck tumor sample into multiple fragments;   (b) culturing the multiple fragments in the presence of at least one cytokine;   (c) obtaining T-cells from the cultured fragments;   (d) expanding the number of T-cells to produce an expanded population of HPV-specific T-cells using
 one or both of (i) irradiated allogenic feeder cells and (ii) irradiated autologous feeder cells; and 
 one or both of (iii) OKT3 antibody and (iv) interleukin-2; 
   (e) optionally, adding a second culturing step following (c); and   (f) administering the population of head and neck tumor-specific T-cells to the mammal in an amount effective to treat or prevent the head and neck cancer in the mammal.   
     
     
         21 . The method of  claim 20 , wherein the cancer is head and neck squamous-cell carcinoma (HNSCC). 
     
     
         22 . The method of  claim 20 , wherein the cancer is HPV-positive oropharyngeal cancer. 
     
     
         23 . The method of  claim 20 , wherein the cancer is selected from the group consisting of laryngeal cancer, hypopharyngeal cancer, nasal cavity cancer, nasopharyngeal cancer, oral cancer, oropharyngeal cancer, and salivary gland cancer. 
     
     
         24 . The method of  claim 20 , wherein a second culturing step is added after step (c) to further expand the number of selected T-cells. 
     
     
         25 . The method of  claim 20 , wherein the cytokine in step (b) is IL-2. 
     
     
         26 . The method of  claim 20 , wherein the head and neck tumor is HPV + . 
     
     
         27 . The method of  claim 20 , wherein the population of cells obtained in step (f) are enriched for HPV-specific T-cells. 
     
     
         28 . The method of  claim 27 , wherein the population of HPV-specific T-cells recognizes HPV 16-positive cancer cells. 
     
     
         29 . The method of  claim 27 , wherein the population of HPV-specific cells recognizes an HPV antigen selected from the group consisting of HPV 16 E6 and HPV 16 E7. 
     
     
         30 . The method of  claim 27 , wherein the population of HPV-specific cells recognizes an HPV antigen selected from the group consisting of HPV 18 E6 and HPV 18 E7. 
     
     
         31 . The method of  claim 20 , further comprising administering to the mammal nonmyleoablative lymphodepleting chemotherapy prior to step (f). 
     
     
         32 . A method of treating or preventing head and neck squamous-cell carcinoma in a human, the method comprising:
 (a) dividing a head and neck squamous-cell carcinoma tumor sample into multiple fragments;   (b) culturing the multiple fragments in the presence of at least one cytokine;   (c) obtaining T-cells from the cultured fragments;   (d) expanding the number of T-cells to produce an expanded population of HPV-specific T-cells using
 one or both of (i) irradiated allogenic feeder cells and (ii) irradiated autologous feeder cells; and 
 one or both of (iii) OKT3 antibody and (iv) interleukin-2; 
   (e) optionally, adding a second culturing step following (c);   (f) administering to the human nonmyleoablative lymphodepleting chemotherapy, and   (g) administering the population of head and neck squamous-cell carcinoma tumor-specific T-cells to the human in an amount effective to treat or prevent the head and neck squamous cell carcinoma in the human.   
     
     
         33 . A method of treating or preventing an HPV+ cancer in a female subject, the method comprising:
 (a) dividing an HPV +  tumor sample into multiple fragments;   (b) culturing the multiple fragments in the presence of at least one cytokine;   (c) obtaining T-cells from the cultured fragments;   (d) expanding the number of T-cells to produce an expanded population of HPV-specific T-cells using
 one or both of (i) irradiated allogenic feeder cells and (ii) irradiated autologous feeder cells; and 
 one or both of (iii) OKT3 antibody and (iv) interleukin-2; 
   (e) optionally, adding a second culturing step following (c); and   (f) administering the population of HPV +  tumor-specific T-cells to the female subject in an amount effective to treat or prevent the HPV+ cancer in the female subject.   
     
     
         34 . The method of  claim 33 , wherein the HPV +  cancer is cervical cancer. 
     
     
         35 . A method of treating or preventing HPV+ cervical cancer in a human female, the method comprising:
 (a) dividing an HPV +  cervical cancer tumor sample into multiple fragments;   (b) culturing the multiple fragments in the presence of at least one cytokine;   (c) obtaining T-cells from the cultured fragments;   (d) expanding the number of T-cells to produce an expanded population of HPV-specific T-cells using one or both of (i) irradiated allogenic feeder cells and (ii) irradiated autologous feeder cells; and   one or both of (iii) OKT3 antibody and (iv) interleukin-2;   (e) optionally, adding a second culturing step following (c); and   (f) administering the population of HPV +  cervical cancer tumor-specific T-cells to the human female in an amount effective to treat or prevent the HPV+ cervical cancer in the human female.   
     
     
         36 . The method of  claim 33 , further comprising administering to the female subject nonmyleoablative lymphodepleting chemotherapy prior to step (f). 
     
     
         37 . The method of  claim 35 , further comprising administering to the human female nonmyleoablative lymphodepleting chemotherapy prior to step (f).