Histidine-tagged intimin and methods of using intimin to stimulate an immune response and as an antigen carrier with targeting capability

Abstract

The present invention describes the isolation and purification of histidine-tagged functional portions of intimin (his-tagged intimin or his-intimin), a protein associated with the ability of certain strains of pathogenic bacteria to adhere to epithelial cells. The invention further describes the use of intimin as an antigen to promote a protective immune response. In addition, the invention describes the combination of intimin with one or more other antigens and administration of the combination to promote a protective immune response against intimin and the one or more antigens. One aspect of the invention is the administration of intimin to target specific epithelial cells to promote a protective immune response to intimin proteins. Additional aspects of the invention include the use of intimin or intimin combined with one or more antigens and administration of the combination to target gastrointestinal mucosa and stimulate an immune response. Additionally, the invention describes administration of the combination of intimin combined with drugs, to provide a means for targeted delivery of drugs to specific epithelial cells. Other aspects of the invention include the production of antibodies directed against his-intimin and methods of using such antibodies to provide passive immune protection, and in an assay system.

1 - 14 . (canceled)  
     
     
         15 . A method of promoting a protective immune response against bacteria expressing intimin or intimin-like proteins, comprising administering to a patient intimin, and intimin-like protein, or a portion thereof which induces antibodies that block binding activity.  
     
     
         16 . A method according to  claim 15 , wherein said intimin, intimin-like protein, or portion thereof is purified.  
     
     
         17 . A method according to  claim 15 , wherein said intimin, intimin-like protein, or portion thereof is enriched.  
     
     
         18 . A method according to  claim 15 , wherein said intimin, intimin-like protein, or portion thereof is histidine-tagged.  
     
     
         19 . A method of promoting a protective immune response against at least one antigen comprising administering to a patient a composition comprising at least one antigen chemically, physically or recombinantly conjugated to intimin, to an intimin-like protein, or to a portion thereof, wherein said portion retains binding function.  
     
     
         20 . A method according to  claim 19 , wherein said intimin, intimin-like protein, or portion thereof is purified.  
     
     
         21 . A method according to  claim 19 , wherein said intimin, intimin-like protein, or portion thereof is enriched.  
     
     
         22 . A method according to  claim 19 , wherein said intimin, intimin-like protein, or portion thereof is histidine-tagged.  
     
     
         23 . A method of targeting the delivery of at least one antigen, at least one drug, or a combination thereof to epithelial cells, comprising administering to a patient a composition comprising at least one antigen, at least one drug or a combination thereof, conjugated to intimin, to an intimin-like protein, or to a portion thereof, wherein said portion retains binding function.  
     
     
         24 . A method according to  claim 23 , wherein said intimin, intimin-like protein, or portion thereof is purified.  
     
     
         25 . A method according to  claim 23 , wherein said intimin, intimin-like protein, or portion thereof is enriched.  
     
     
         26 . A method according to  claim 23 , wherein said intimin, intimin-like protein, or portion thereof is histidine-tagged.  
     
     
         27 . A method of providing passive immune protection comprising administering anti-intimin antibodies to a patient in need thereof.  
     
     
         28 - 46 . (canceled)  
     
     
         47 . A method of promoting a protective immune response against bacteria expressing intimin or intimin-like proteins, comprising administering to a patient intimin, and intimin-like protein, or a portion thereof which retains binding activity.  
     
     
         48 . A method according to  claim 47 , wherein said intimin, intimin-like protein, or portion thereof is purified.  
     
     
         49 . A method according to  claim 47 , wherein said intimin, intimin-like protein, or portion thereof is enriched.  
     
     
         50 . A method according to  claim 47 , wherein said intimin, intimin-like protein, or portion thereof is histidine-tagged.