Induction of a Th1-like response in vitro

Abstract

The invention provides compositions and methods for stimulating a Th1-like response in vitro. Compositions include fusion proteins and conjugates that contain at least a portion of a heat shock protein. A Th1-like response can be elicited by contacting in vitro a cell sample containing naive lymphocytes with a fusion protein or conjugate of the invention. The Th1-like response can be detected by measuring IFN-gamma produced by the cell sample.

1 - 64 . (canceled)  
     
     
         65 . A method of determining whether a fusion protein stimulates a Th1-like response, the method comprising: 
 (a) providing a cell sample comprising naive lymphocytes in vitro;    (b) providing a fusion protein comprising (i) a heat shock protein (Hsp) or a fragment thereof at least eight amino acid residues in length, and (ii) an amino acid sequence containing an MHC class I or MHC class II binding epitope of (a) a tumor associated antigen, or (b) a protein produced by a human pathogen;    (c) contacting the cell sample with the fusion protein; and    (d) determining whether the fusion protein stimulates a Th1-like response in the cell sample.    
     
     
         66 . The method of  claim 65 , wherein the Hsp is selected from the group consisting of Hsp65, Hsp40, Hsp10, Hsp60, and Hsp71.  
     
     
         67 . The method of  claim 65 , wherein the fusion protein comprises a full length Hsp selected from the group consisting of Hsp65, Hsp40, Hsp10, Hsp60, and Hsp71.  
     
     
         68 . The method of  claim 65 , wherein the fusion protein comprises amino acids 1-200 of Hsp65 of  Mycobacterium bovis.    
     
     
         69 . The method of  claim 65 , wherein the fusion protein comprises an amino acid sequence containing an MHC class I or MHC class II binding epitope of a protein produced by a virus.  
     
     
         70 . The method of  claim 69 , wherein the virus is selected from the group consisting of human papilloma virus (HPV), herpes simplex virus (HSV), hepatitis B virus (HBV), hepatitis C virus (HCV), cytomegalovirus (CMV), Epstein-Barr virus (EBV), influenza virus, measles virus, and human immunodeficiency virus (HIV).  
     
     
         71 . The method of  claim 65 , wherein the fusion protein comprises an HPV E7 protein or an MHC class I or MHC class II binding epitope of an HPV E7 protein.  
     
     
         72 . The method of  claim 65 , wherein the fusion protein comprises an HPV E6 protein or an MHC class I or MHC class II binding epitope of an HPV E6 protein.  
     
     
         73 . The method of  claim 65 , wherein the fusion protein comprises an HPV type 16 E7 protein or an MHC class I or MHC class II binding epitope of an HPV type 16 E7 protein.  
     
     
         74 . The method of  claim 65 , wherein the fusion protein comprises an HPV type 16 E6 protein or an MHC class I or MHC class II binding epitope of an HPV type 16 E6 protein.  
     
     
         75 . The method of  claim 65 , wherein the fusion protein comprises a  Mycobacterium bovis  Hsp65 protein and an HPV type 16 E7 protein.  
     
     
         76 . The method of  claim 65 , wherein the fusion protein comprises a full length Hsp.  
     
     
         77 . The method of  claim 65 , wherein the fusion protein comprises a fragment of an Hsp that is at least 50 amino acids in length.  
     
     
         78 . The method of  claim 65 , wherein the fusion protein comprises a fragment of an Hsp that is at least 100 amino acids in length.  
     
     
         79 . The method of  claim 65 , wherein the fusion protein comprises a fragment of an Hsp that is at least 200 amino acids in length.  
     
     
         80 . The method of  claim 65 , wherein the fusion protein comprises a fragment of an Hsp that is at least 300 amino acids in length.  
     
     
         81 . The method of  claim 65 , wherein the cell sample comprises cells derived from a spleen, lymph node, peripheral blood, bone marrow, thymus, lung, respiratory tract, or anogenital mucosa.  
     
     
         82 . The method of  claim 65 , wherein the cell sample comprises splenocytes or lymph node cells.  
     
     
         83 . The method of  claim 65 , wherein the detecting step comprises detecting IFN-gamma produced by the cell sample.  
     
     
         84 . The method of  claim 65 , comprising the further steps of: 
 (e) providing a second cell sample comprising naive lymphocytes;    (f) contacting the second cell sample with a second fusion protein; and    (g) determining whether the second fusion protein stimulates a Th1-like response in the second cell sample,    wherein the first fusion protein comprises the sequence of a full-length, naturally occurring Hsp, and the second fusion protein comprises at least eight amino acids but less than all of the sequence of a naturally occurring Hsp.