Methods for recovering peptides from stress protein-peptide complexes

Abstract

Disclosed is a method for inhibiting the proliferation of a tumor in a mammal. The method involves the steps of (a) isolating a stress protein-peptide complex from tumor cells previously removed from the mammal and (b) administering the isolated stress protein-peptide complex back to the mammal in order to stimulate in the mammal an immune response against the tumor from which the complex was isolated. Stress protein-peptide complexes having particular utility in the practice of the instant invention include the Hsp70-peptide, Hsp90-peptide and gp96-peptide complexes.

1 . A method for inhibiting proliferation of a tumor in a mammal, the method comprising: 
 administering to the mammal harboring the tumor a composition comprising, 
 (a) an immunogenic stress protein-peptide complex isolated from a cell derived from the tumor, said complex being operative to initiate in the mammal an immune response against said tumor, and  
 (b) a pharmaceutically acceptable carrier,  
   in an amount sufficient to elicit in the mammal an immune response against the tumor thereby inhibiting proliferation of the tumor.    
     
     
         2 - 18 . (canceled)  
     
     
         19 . A method of preparing a composition comprising a recovered population of peptides in admixture with a pharmaceutically acceptable nontoxic carrier, wherein said method comprises the steps of: 
 (a) purifying a population of stress protein-peptide complexes from mammalian tumor cells, wherein the stress protein is noncovalently associated with the peptide in said complexes;    (b) releasing the peptides from said population of complexes to produce a released population of peptides;    (c) recovering the released population of peptides; and    (d) combining said released population of peptides with a pharmaceutically acceptable nontoxic carrier.    
     
     
         20 . The method of  claim 19 , wherein the peptides are released from said population of complexes by a method comprising placing said population of complexes in the presence of adenosine triphosphate.  
     
     
         21 . The method of  claim 19 , wherein said mammalian tumor cells are human cells.  
     
     
         22 . The method of  claim 19 , wherein said mammalian tumor cells are from a tumor selected from the group consisting of melanocarcinoma, hepatocarcinoma, and renal cell carcinoma.  
     
     
         23 . The method of  claim 19 , wherein said tumor cells are from a metastasis.  
     
     
         24 . The method of  claim 19 , wherein said tumor cells have been proliferated in vivo.  
     
     
         25 . The method of  claim 19 , wherein said tumor cells have been proliferated in vitro.  
     
     
         26 . The method of  claim 19 , wherein the stress protein is a member of a stress protein family selected from the group consisting of hsp60, hsp70 and hsp90.  
     
     
         27 . The method of  claim 19 , wherein the stress protein is hsp70.  
     
     
         28 . The method of  claim 19 , wherein the stress protein is hsp90.  
     
     
         29 . The method of  claim 19 , wherein the stress protein is gp96.  
     
     
         30 . The method of  claim 19 , further comprising combining said released population of peptides with an adjuvant.  
     
     
         31 . The method of  claim 30 , wherein the adjuvant is selected from the group consisting of a pluronic tri-block copolymer, muramyl dipeptide, detoxified endotoxin, saponin, QS-21, and liposome.