Il-12 immunotherapy for cancer

Abstract

Compositions and methods for delivering immune modulatory molecules to result in a therapeutic effect are disclosed. The compositions and methods use stably integrating lentiviral delivery systems. The methods are useful for therapeutically and prophylactically treating cancer such as leukemia.

1 - 73 . (canceled) 
     
     
         74 . A lentiviral vector comprising an IL-12 expression cassette, wherein the IL-12 expression cassette comprises a polynucleotide encoding a p35 polypeptide and a p40 polypeptide, wherein the polynucleotide has at least 90% sequence identity to SEQ ID NO: 20 or SEQ ID NO: 21 and the polypeptide encoded by the polynucleotide activates an IL-12 receptor. 
     
     
         75 . The lentiviral vector of  claim 74 , wherein the lentiviral vector further comprises a central polypurine tract (cPPT), a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE), a 5′-long terminal repeat (LTR), a HIV signal sequence, a HIV Psi signal 5′-splice donor (SD), a delta-GAG element, a rev response element (RRE), a 3′-splice acceptor (SA), and a 3′-self-inactivating (SIN) LTR. 
     
     
         76 . A method of treating cancer in a human patient, the method comprising administering to the patient a pharmaceutical composition comprising (i) a population of cells that secrete interleukin 12 (IL-12), wherein the cells comprise an IL-12 expression cassette, and (ii) a pharmaceutically acceptable excipient, diluent, or carrier, wherein the cells are obtained by:
 a. contacting isolated human cells with a viral vector comprising the IL-12 expression cassette, thereby transducing the isolated human cells to express IL-12;   b. detecting the concentration of IL-12 secreted by the human cells transduced in (a) when cultured at a density of 10 6  cells/ml for two hours; and   c. releasing the transduced human cells for treatment of cancer in the patient if the concentration of secreted IL-12 detected in (b) is at least 1,500 pg/ml.   
     
     
         77 . The method of  claim 76 , wherein the viral vector is a retroviral vector. 
     
     
         78 . The method of  claim 77 , wherein the retroviral vector is a lentiviral vector. 
     
     
         79 . The method of  claim 78 , wherein the lentiviral vector comprises a cPPT, a WPRE, a 5′-LTR, a HIV signal sequence, a HIV Psi signal 5′-SD, a delta-GAG element, a RRE, a 3′-SA, and a 3′-SIN-LTR. 
     
     
         80 . The method of  claim 76 , wherein the human cells are cancer cells. 
     
     
         81 . The method of  claim 80 , wherein the cancer cells are leukemia cells. 
     
     
         82 . The method of  claim 76 , wherein the human cells are T-cells. 
     
     
         83 . The method of  claim 76 , wherein the human cells are stem cells.