Stress protein-peptide complexes as prophylactic and therapeutic vaccines against intracellular pathogens

Abstract

Disclosed is a family of vaccines that contain stress protein-peptide complexes which when administered to a mammal are operative to initiate in the mammal a cytotoxic T cell response against cells infected with a preselected intracellular pathogen. Also disclosed are methodologies for preparing and administering vaccines containing such stress protein-peptide complexes.

What is claimed is:  
     
         1 . A vaccine for administration to a mammal for inducing in the mammal a cytotoxic T cell response against a preselected intracellular pathogen, the vaccine comprising: 
 (a) an immunogenic stress protein-peptide complex operative to initiate in said mammal a cytotoxic T cell response against said pathogen, said complex comprising, 
 a peptide that is present in a eukaryotic cell infected with said pathogen but not present in said cell when said cell is not infected with said pathogen, complexed with a stress protein; and  
   (b) a pharmaceutically acceptable carrier.    
     
     
         2 . A vaccine for administration to a mammal for inducing in said mammal resistance to infection by a preselected intracellular pathogen, the vaccine comprising: 
 (a) an immunogenic stress protein-peptide complex operative to initiate in said mammal, by means of a cytotoxic T cell response in said mammal, resistance to infection by said pathogen, said complex comprising, 
 a peptide that is present in a eukaryotic cell infected with said pathogen but not present in said cell when said cell is not infected with said pathogen, complexed with a stress protein; and  
   (b) a pharmaceutically acceptable carrier.    
     
     
         3 . The composition of  claim 1  or  2 , wherein said stress protein is a member of the stress protein families selected from the group consisting of Hsp60, Hsp70, and Hsp90.  
     
     
         4 . The composition of  claim 1  or  2 , wherein said stress protein is a gp96.  
     
     
         5 . The composition of  claim 1  or  2  further comprising a cytokine.  
     
     
         6 . The composition of  claim 5 , wherein said cytokine is selected from the group consisting of IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IFNα, IFNβ, IFNγ, TNFα, TNFβ, G-CSF, GM-CSF, GM-CSF and TGF-β.  
     
     
         7 . The composition of  claim 1  or  2 , wherein said eukaryotic cell is an immortalized eukaryotic cell.  
     
     
         8 . The composition of  claim 1  or  2 , wherein said mammal is a human.  
     
     
         9 . The composition of  claim 1  or  2 , wherein said pathogen is a virus.  
     
     
         10 . The composition of  claim 9 , wherein said virus is selected from the group consisting of hepatitis type A, hepatitis type B, hepatitis type C, influenza, varicella, adenovirus, herpes simplex type I, herpes simplex type II, rinderpest, rhinovirus, echovirus, rotavirus, respiratory synctial virus, papilloma virus, papova virus, cytomegalovirus, echinovirus, arbovirus, huntavirus, coxsachie virus, mumps virus, measles virus, rubella virus, polio virus, human immunodeficiency virus type I, and human immunodeficiency virus type II.  
     
     
         11 . The composition of  claim 1  or  2 , wherein said pathogen is a bacteria.  
     
     
         12 . The composition of  claim 11 , wherein said bacteria is selected from the group consisting of Mycobacteria, Rickettsia, Neisseria and Legionella.  
     
     
         13 . The composition of  claim 1  or  2 , wherein said pathogen is a protozoa.  
     
     
         14 . The composition of  claim 13 , wherein said protozoa is selected from the group consisting of Leishmania, Trypanosoma and Kokzidioa.  
     
     
         15 . The composition of  claim 1  or  2 , wherein said pathogen is an intracellular parasite.  
     
     
         16 . The composition of  claim 15 , wherein said parasite is selected from the group consisting of Chlamydia and Rickettsia.  
     
     
         17 . The composition of  claim 1  or  2 , wherein said peptide is non covalently complexed with said stress protein.  
     
     
         18 . A method of inducing in a mammal a cytotoxic T cell response against a preselected intracellular pathogen that causes disease in said mammal, the method comprising: 
 administering to said mammal a vaccine comprising, 
 (a) an immunogenic stress protein-peptide complex operative to initiate in said mammal a cytotoxic T cell response against said pathogen and comprising, a peptide that is present in a eukaryotic cell infected with said pathogen but not present in said cell when said cell is not infected with said pathogen, complexed with a stress protein, and  
 (b) a pharmaceutically acceptable carrier,  
   in an amount sufficient to elicit in said mammal a cytotoxic T cell response against said pathogen.    
     
     
         19 . A method of inducing in a mammal resistance to infection by a preselected intracellular pathogen that-causes disease in said mammal, the method comprising: 
 administering to said mammal a vaccine comprising, 
 (a) an immunogenic stress protein-peptide complex operative to initiate in said mammal cytotoxic T cell response against said pathogen and comprising, 
 a peptide that is present in a eukaryotic cell infected with said pathogen but not present in said cell when said cell is not infected with said pathogen, complexed with a stress protein, and  
 
 (b) a pharmaceutically acceptable carrier, in an amount sufficient to induce in said mammal, by means of the cytotoxic T cell response in said mammal, resistance to infection by said pathogen.  
   
     
     
         20 . The method of  claim 18  or  19 , wherein said cytotoxic T cell response is mediated by the class I major histocompatibility complex.  
     
     
         21 . The method of  claim 18  or  19 , wherein said stress protein is a member of the stress protein families selected from the group consisting of Hsp60, Hsp70, and Hsp90.  
     
     
         22 . The method of  claim 18  or  19 , wherein said stress protein is a gp96.  
     
     
         23 . The method of  claim 18  or  19 , wherein said composition further comprises a cytokine.  
     
     
         24 . The method of  claim 23 , wherein said cytokine is selected from the group consisting of IL-1α IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IFNα, IFNβ, IFNγ, TNFα, TNFβ, G-CSF, GM-CSF, GM-CSF and TGF-β.  
     
     
         25 . The method of  claim 18  or  19 , wherein said eukaryotic cell is an immortalized eukaryotic cell.  
     
     
         26 . The method of  claim 18  or  19 , wherein said mammal is a human.  
     
     
         27 . The method of  claim 18  or  19 , wherein said vaccine is administered prophylactically to said mammal for stimulating in said mammal a cytotoxic T cell response for preventing subsequent infection of said mammal by said pathogen.  
     
     
         28 . The method of  claim 18  or  19 , wherein said vaccine is administered therapeutically to said mammal for stimulating in said mammal a cytotoxic T cell response against said pathogen presently infecting said mammal.  
     
     
         29 . The method of  claim 18  or  19 , wherein said vaccine is administered to said mammal in an amount in the range of about 0.1 to about 1000 micrograms of complex/kg body weight of mammal/immunization.  
     
     
         30 . The method of  claim 29 , wherein said amount is in range of about 0.5 to about 100 micrograms of complex/kg body weight of mammal/immunization.  
     
     
         31 . A method for preparing a vaccine for inducing in a mammal a cytotoxic T cell response against a preselected intracellular pathogen, the method comprising: 
 (a) harvesting from a eukaryotic cell infected with said pathogen an immunogenic stress protein-peptide complex comprising, 
 a peptide that is present in said cell infected with said pathogen but not present in said cell when said cell is not infected with said pathogen complexed with a stress protein, said complex, when administered to said mammal, being operative at initiating in said mammal a cytotoxic T cell response against said pathogen; and  
   (b) combining said complex with a pharmaceutically acceptable carrier.    
     
     
         32 . A method for preparing a vaccine for inducing in a mammal a cytotoxic T cell response against a preselected intracellular pathogen, the method comprising: 
 (a) reconstituting in vitro a peptide that is present in a eukaryotic cell infected with said pathogen but not present in said cell when said cell is not infected with said pathogen and a stress protein, thereby to generate a stress protein-peptide complex, which when administered to said mammal is operative to initiate a cytotoxic T cell response against said pathogen in said mammal; and    (b) combining said complex with a pharmaceutically acceptable carrier.    
     
     
         33 . The method of  claim 32 , wherein said stress protein is harvested in the presence of ATP prior to reconstitution.  
     
     
         34 . The method of  claim 32 , wherein said stress protein is treated with loss pH prior to reconstitution.  
     
     
         35 . The method of  claim 31  or  32 , wherein said stress protein is a member of the stress protein families selected from the group consisting of Hsp60, Hsp70 and Hsp90.  
     
     
         36 . The method of  claim 31  or  32 , wherein said stress protein is a gp96.