Cryopreservation of apoptotic cancer cells for use in immunotherapy against cancer

Abstract

Described herein is a reliable method for preparing a potent vaccine useful for immunotherapy comprising the step of cryopreserving a population of cells undergoing immunogenic cell death, and using such cells to activate dendritic cells for use in immunotherapy. In a specific embodiment, the method comprises cryopreserving cancer cells undergoing cell death, which can be used to prepare a pharmaceutical composition for immunotherapy against cancer.

1 - 24 . (canceled) 
     
     
         25 . A method for preparing a pharmaceutical composition for use in immunotherapy comprising:
 a) inducing immunogenic cell death in a population of cells expressing an antigen of interest;   b) cryopreserving the cells undergoing immunogenic cell death in a cryopreservant;   c) thawing the cryopreserved cells;   d) pulsing immature dendritic cells with the thawed cryopreserved cells; and   e) co-culturing the pulsed dendritic cells with lymphocytes.   
     
     
         26 . The method of  claim 25 , wherein the cells expressing an antigen of interest are obtained from one or more cell line. 
     
     
         27 . The method of  claim 26 , wherein the one or more cell line is a cancer cell line. 
     
     
         28 . The method of  claim 25 , wherein the lymphocytes are T lymphocytes. 
     
     
         29 . The method of  claim 25 , wherein the cells undergoing immunogenic cell death are cryopreserved within 2.5 hours after induction of immunogenic cell death. 
     
     
         30 . The method of  claim 25 , wherein the immunogenic cell death is induced by high hydrostatic pressure, anthracyclines, anti-EGFR antibodies, Big Potassium channel antagonists, bortezomib, cardiac glycosides, cyclophosphamide, GADD43/PP1 inhibitors and mitomycin, irradiation by UV light or gamma rays, oxaliplatin, photodynamic therapy with hypericin, poly(I:C), or thapsigargin and cisplatin. 
     
     
         31 . A method for administering immunotherapy to a patient comprising:
 a) inducing immunogenic cell death in a population of cells expressing an antigen of interest;   b) cryopreserving the cells undergoing immunogenic cell death in a cryopreservant;   c) thawing the cryopreserved cells;   d) pulsing immature dendritic cells obtained from the patient with the thawed cryopreserved cells;   e) co-culturing the pulsed dendritic cells with lymphocytes obtained from the patient; and   f) administering the lymphocytes co-cultured with the pulsed dendritic cells to the patient.   
     
     
         32 . The method of  claim 31 , wherein the cells expressing an antigen of interest are obtained from one or more cell line. 
     
     
         33 . The method of  claim 32 , wherein the one or more cell line is a cancer cell line. 
     
     
         34 . The method of  claim 31 , wherein the lymphocytes are T lymphocytes. 
     
     
         35 . The method of  claim 31 , wherein the immunogenic cell death is induced by high hydrostatic pressure, anthracyclines, anti-EGFR antibodies, Big Potassium channel antagonists, bortezomib, cardiac glycosides, cyclophosphamide, GADD43/PP1 inhibitors and mitomycin, irradiation by UV light or gamma rays, oxaliplatin, photodynamic therapy with hypericin, poly(I:C), or thapsigargin and cisplatin.