Antigen presenting cells, method for their preparation and their use for cancer vaccines

Abstract

The present invention discloses a method of generation of antigen presenting cells, comprising: a. collecting said cells from a subject, b. activating said collected cells; c. culturing and optionally expanding ex vivo said activated cells; d. treating said cultured and optionally expanded cells with DNA hypomethylating agents so that said cells concomitantly express multiple tumor associated antigens. The cells obtainable according to the method of the present invention, as well as the cellular components thereof whether alone or in combination with said cells, are useful for prevention and treatment of malignancies of different histotype that constitutively express one or more of the multiple tumor associated antigens that are expressed in said cells. Conveniently, said cells and/or cellular components are in the form of a vaccine. Said vaccines are advantageous over the prior art in that as they concomitantly express multiple/all methylation-regulated tumor associated antigens.

1 - 31 . (Canceled).  
     
     
         32 . A method for the generation of cells presenting cancer-testis antigens comprising: 
 a) collecting peripheral blood mononuclear cells from a subject or cells derived from peripheral blood mononuclear cells,    b) activating said collected cells to form activated cells;    c) culturing and optionally expanding ex vivo said activated cells;    d) treating said cultured and optionally expanded cells with DNA hypomethylating agents by pulsing said cells four times every 12 hours; then replacing half of the culture medium with fresh medium and allowing to proceed for additional 48 hours so that said cells express said antigens.    
     
     
         33 . A method according to  claim 32 , wherein said subject is a mammal.  
     
     
         34 . A method according to  claim 33 , wherein said subject is a human.  
     
     
         35 . A method according to  claim 32 , wherein said subject is a cancer patient.  
     
     
         36 . A method according to  claim 32 , wherein said activated cells are Epstein-Barr virus-immortalized B-lymphoblastoid cell lines.  
     
     
         37 . A method according to  claim 32 , wherein said activated cells are Pokeweed mitogen (PWM)-activated B-lymphocytes.  
     
     
         38 . A method according to  claim 32 , wherein said activated cells are CD40 activated B-lymphocytes.  
     
     
         39 . A method according to  claim 32 , wherein said activated cells are Phytohemagglutinin (PHA)+recombinant human interleukin-2 (rhIL-2)-activated PBMC.  
     
     
         40 . A method according to  claim 32 , wherein said activated cells are Phytohemagglutinin (PHA)+recombinant human interleukin-2 (rhIL-2)+pokeweed mitogen (PWM)-activated PBMC.  
     
     
         41 . A method according to  claim 32 , wherein histone deacetylase inhibitors are further used in step d).  
     
     
         42 . A method according to  claim 32 , wherein said DNA hypomethylating agent is selected from 5-aza-cytidine or 5-aza-2′-deoxycytidine.  
     
     
         43 . Cells obtainable by the method according to any of claims  32 - 42  and  62 - 65 .  
     
     
         44 . A method according to  claim 32 , wherein said subject is a healthy subject.  
     
     
         45 . A method according to  claim 37 , wherein said activation is carried out for 2 days.  
     
     
         46 . A method according to  claim 39 , wherein said activation is carried out for 2 days.  
     
     
         47 . A method according to  claim 40 , wherein said activation is carried out for 2 days.