Mycobacterium Antigens

Abstract

There is provided a diagnostic reagent for use in the detection of M. bovis or M. tuberculosis infection in an animal, comprising a peptide which has an epitope from Mycobacterium bovis hypothetic protein Mb3645c (SEQ ID NO: 1) or an epitope from a polypeptide having at least 76% identity with SEQ ID NO: 1.

1 : A diagnostic reagent for use in the detection of  M. tuberculosis  infection in a human, comprising a peptide having an amino acid sequence at least 90% identical to SEQ ID NO: 1, said peptide comprising an epitope consisting of amino acids from within SEQ ID NO: 1, wherein said peptide has fewer than 103 amino acids. 
     
     
         2 : The diagnostic reagent according to  claim 1 , wherein said epitope is a T-cell epitope. 
     
     
         3 : The diagnostic reagent according to  claim 1 , wherein said diagnostic reagent is a cell-mediated immunity (CMI) assay diagnostic reagent. 
     
     
         4 : The diagnostic reagent according to  claim 1 , wherein said epitope is a T-cell epitope, and wherein said diagnostic reagent is a cell-mediated immunity (CMI) assay diagnostic reagent. 
     
     
         5 : The diagnostic reagent according to  claim 1 , wherein said peptide is a truncated form of SEQ ID NO: 1. 
     
     
         6 : The diagnostic reagent according to  claim 1 , wherein said epitope has 5 or more amino acids. 
     
     
         7 : The diagnostic reagent according to  claim 1 , further comprising one or more peptides each selected from amino acid sequences SEQ ID NO:14-25. 
     
     
         8 : The diagnostic reagent according to  claim 1 , further comprising one or more peptides each selected from amino acid sequences SEQ ID NO:16-25. 
     
     
         9 : The diagnostic reagent according to  claim 1 , further comprising one or more peptides each selected from amino acid sequences SEQ ID NO:14-25 and one or more peptides each selected from amino acid sequences SEQ ID NO:16-25. 
     
     
         10 : A diagnostic reagent for use in the detection of  M. tuberculosis  infection in a human, comprising a peptide having an amino acid sequence at least 90% identical to SEQ ID NO: 1, said peptide comprising an epitope consisting of amino acids from within SEQ ID NO: 1, wherein said peptide has 100 amino acids or fewer. 
     
     
         11 : The diagnostic reagent according to  claim 10 , wherein said peptide has 75 amino acids or fewer. 
     
     
         12 : The diagnostic reagent according to  claim 11 , wherein said peptide has 50 amino acids or fewer. 
     
     
         13 : The diagnostic reagent according to  claim 12 , wherein said peptide has 25 amino acids or fewer. 
     
     
         14 : The diagnostic reagent according to  claim 13 , wherein said peptide has 20 amino acids or fewer. 
     
     
         15 : The diagnostic reagent according to  claim 10 , wherein said epitope is a T-cell epitope, and wherein said diagnostic reagent is a cell-mediated immunity (CMI) assay diagnostic reagent. 
     
     
         16 : The diagnostic reagent according to  claim 10 , wherein said epitope has 5 or more amino acids. 
     
     
         17 : The diagnostic reagent according to  claim 10 , further comprising one or more peptides each selected from amino acid sequences SEQ ID NO:14-25 or one or more peptides each selected from amino acid sequences SEQ ID NO:16-25. 
     
     
         18 : The diagnostic reagent according to  claim 17 , further comprising one or more peptides each selected from amino acid sequences SEQ ID NO:14-25 and one or more peptides each selected from amino acid sequences SEQ ID NO:16-25. 
     
     
         19 : A method for diagnosing, in a human, infection by or exposure to  M. tuberculosis  versus prior  M. bovis bacillus  Calmette-Guerin (BCG) vaccination, comprising the steps of:
 (a) contacting a population of cells from said human with a diagnostic reagent comprising a peptide having an amino acid sequence at least 90% identical to SEQ ID NO: 1, said peptide comprising an epitope consisting of amino acids from within SEQ ID NO: 1; and   (b) determining whether cells of said population of cells recognize the epitope;   wherein recognition by said population of cells of said epitope indicates infection by or exposure to  M. tuberculosis  irrespective of prior  M. bovis bacillus  Calmette-Guerin (BCG) vaccination.   
     
     
         20 : The method according to  claim 19 , wherein determining step (b) comprising comparing IFNγ levels to a control. 
     
     
         21 : A method of diagnosing  M. tuberculosis  infection in a human, or of determining whether a human has been exposed to  M. tuberculosis , comprising:
 (i) contacting T cells from said human with one or more of
 (a) a peptide having the sequence shown in SEQ ID NO: 1 
 (b) a peptide having a sequence of at least 8 consecutive amino acids of the sequence shown in SEQ ID NO: 1; or 
 (c) a peptide having a sequence that is capable of binding to a T-cell receptor which recognizes a peptide having the sequence shown in SEQ ID NO: 1 or a sequence of at least 8 consecutive amino acids of the sequence shown in SEQ ID NO: 1; and 
   (ii) determining whether the said T-cells recognize said peptide, wherein recognition of said peptide by said T-cells is determined by detecting a cytokine from the T cells using a cell-mediated immunity (CMI) assay.   
     
     
         22 : The method of  claim 21 , wherein step (i) further comprises contacting said T cells from said human with one or more  M. tuberculosis  antigens. 
     
     
         23 : The method of  claim 22 , wherein said one or more  M. tuberculosis  antigens include antigens encoded by the RD-1 region, which antigens are selected from ESAT-6, CFP10, fragments thereof with SEQ ID NOs: 16-25, or combinations thereof. 
     
     
         24 : The method of  claim 21 , wherein one or more of the peptides:
 (i) is represented by SEQ ID Nos: 2-13, or   (ii) binds to a T-cell that recognizes (i),   are contacted with the T-cells.   
     
     
         25 : The method of  claim 21 , wherein said cytokine is IFN-γ. 
     
     
         26 : The method of  claim 21 , wherein said cytokine is detected by an enzyme-linked immunosorbent assay. 
     
     
         27 : The method of  claim 21 , wherein said T-cells have been cultured in vitro. 
     
     
         28 : The method of  claim 21 , wherein said peptide has fewer than 103 amino acids. 
     
     
         29 : The method of  claim 21 , wherein said peptide has up to 100 amino acids. 
     
     
         30 : A kit for diagnosing  M. tuberculosis  infection or exposure in a human, said kit comprising a composition comprising:
 (a) a peptide having the sequence shown in SEQ ID NO: 1;   (b) a peptide having a sequence of at least 8 consecutive amino acids of the sequence shown in SEQ ID NO: 1;   (c) a peptide having a sequence which is capable of binding to a T-cell receptor which recognizes a peptide defined in (a) or (b); or   (d) at least one peptide:
 (i) which is presented by SEQ ID Nos: 2-13, or 
 (ii) which binds to a T-cell that recognizes (i), 
   and the composition optionally comprises one or more further  M. tuberculosis  T-cell antigens,   wherein the kit comprises a means for detecting recognition of a peptide by a T-cell.   
     
     
         31 : The kit of  claim 30 , wherein said one or more further  M. tuberculosis  T-cell antigens is selected from ESAT-6 and/or CFP10, fragments with SEQ ID NOs: 16-25 or combinations thereof. 
     
     
         32 : The kit of  claim 30 , wherein said means for detecting recognition of said peptide by T-cells is a cell-mediated immunity (CMI) assay. 
     
     
         33 : The kit of  claim 30 , wherein said peptide has fewer than 103 amino acids. 
     
     
         34 : The kit of  claim 30 , wherein said peptide has up to 100 amino acids.