Tuberculosis antigen detection assays and vaccines

Abstract

The present invention relates to isolated Tuberculosis (TB) antigens that are useful in therapeutic and vaccine compositions for stimulating a TB specific immunological response. The identified antigens are also useful in diagnostic assays to determine the presence of active TB in an individual. Accordingly, the present invention includes polypeptide molecules, nucleic acid molecules, vaccine compositions, diagnostic assays, and methods of diagnosis and monitoring treatment related to these TB antigens.

1 . An isolated polypeptide molecule that consists of an immunogenic portion of a  Mycobacterium tuberculosis  antigen, wherein said antigen consisting of:
 a) an amino acid sequence encoded by a nucleic acid molecule of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, or combination thereof;   b) an amino acid sequence encoded by a coding region of a nucleic acid molecule of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, or combination thereof;   c) an amino acid sequence encoded by a complement of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, or combination thereof;   d) an amino acid sequence encoded by a nucleic acid molecule that hybridizes to SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, or combination thereof under high stringency conditions, wherein said conditions comprise 1×SSC, 1% SDS and 0.1-2 mg/ml denatured calf thymus DNA at 65° C.; or   e) an amino acid sequence set forth in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, or combination thereof   
     
     
         2 . The isolated polypeptide molecule of  claim 1 , wherein the isolated polypeptide molecule stimulates an immunogenic specific Tuberculosis Bacterium (TB) response in a host. 
     
     
         3 . An isolated polypeptide molecule that consists of an immunogenic portion of a  M. tuberculosis  antigen, wherein said antigen having greater than or equal to about 70% similarity with a sequence consisting of:
 a) an amino acid sequence encoded by a nucleic acid molecule of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, or combination thereof;   b) an amino acid sequence encoded by a coding region of a nucleic acid molecule of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, or combination thereof;   c) an amino acid sequence encoded by a complement of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, or combination thereof;   d) an amino acid sequence encoded by a nucleic acid molecule that hybridizes to SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, or combination thereof, under high stringency conditions, wherein said conditions comprise 1×SSC, 1% SDS and 0.1-2 mg/ml denatured calf thymus DNA at 65° C.; or   e) an amino acid sequence set forth in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, or combination thereof   
     
     
         4 . The isolated polypeptide molecule of  claim 3 , wherein the isolated polypeptide molecule stimulates an immunogenic specific TB response in a host. 
     
     
         5 . The isolated polypeptide molecule of  claim 3 , wherein the nucleic acid molecule has greater than or equal to about 80% similarity with said sequences. 
     
     
         6 . The isolated polypeptide molecule of  claim 5 , wherein the nucleic acid molecule has greater than or equal to about 90% similarity with said sequences. 
     
     
         7 . A fusion protein comprising a polypeptide selected from the group consisting of:
 a) at least one of the polypeptides of  claim 1 ;   b) at least two of the polypeptides of  claim 1 ;   c) at least one polypeptide of  claim 1  and a known  M. tuberculosis  antigen; and   d) at least one polypeptide of  claim 1  and an  M. tuberculosis  antigen presented on a MHC Class-2 molecule.   
     
     
         8 . A composition that comprises the polypeptide sequence of  claim 1  and a physiologically acceptable carrier. 
     
     
         9 . The composition of  claim 8 , further including an immune response enhancer. 
     
     
         10 . The composition of  claim 9 , wherein the immune response enhancer is an adjuvant or another TB antigen. 
     
     
         11 . The composition of  claim 10 , wherein the composition is a vaccine composition. 
     
     
         12 . The composition of  claim 8 , wherein the adjuvant includes at least one component selected from the group consisting of 3D-MPL and QS21. 
     
     
         13 . The composition of  claim 8 , wherein the composition is formulated in an oil and water emulsion. 
     
     
         14 . The composition of  claim 9 , wherein the immune response enhancer is an immunostimulatory cytokine or chemokine 
     
     
         15 . A kit for diagnosing the presence or absence of  M. tuberculosis  infection in a person, the kit comprises one or more reagents for detecting the polypeptide molecule of  claim 1 . 
     
     
         16 . A pharmaceutical composition comprising the polypeptide of  claim 1 , and a carrier.