Mycobacterium tuberculosis epitopes and methods of use thereof
AssigneeBeckman Coulter, Inc.
InventorMarkus Macurer
Life of this patent
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Abstract
The invention provides novel Class I HLA-A2 and Class II HLA-DR4-restricted epitopes and methods for their use in detecting T-cells in peripheral blood specific for infection or latency of mycobacterial infection, including M. tuberculosis and M. leprae as others. For example, methods for diagnosing the presence of infection or exposure by M. tuberculosis utilize multimers of HLA monomers or modified monomers having a bound HLA-binding peptide to perform high throughput screening of patient PBLs. The methods can be used for monitoring the success of anti mycobacterial treatment in patients and to screen vaccines and drugs for effectiveness in treating or preventing exposure, infection and latency of mycobacteria in humans.
Claims36
An isolated human T-cell epitope derived from Mycobacterium tuberculosis (MTB), said epitope comprising an amino acid sequence as set forth in SEQ ID NO:3, 6, 7 or 8.
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A method for diagnosing the presence of a infection by or exposure to MTB in a human subject, comprising:
- a) contacting PBLs obtained from the subject with multimers or oligomers of HLA-DR4 monomers or modified monomers, having a bound HLA-binding peptide comprising an amino acid sequence as set forth in SEQ ID NO:8,
- b) detecting binding of the multimers or oligomers to the PBLs wherein the binding to PBLs is indicative of the presence of MTB infection or exposure in the subject.
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A method for diagnosing the presence of an infection by or exposure to MTB in a human subject, comprising:
- a) contacting PBLs obtained of the subject with tetramers of HLA-A2 monomers, or modified monomers, oligomer or multimers having a bound HLA-binding peptide comprising an amino acid sequence as set forth in SEQ ID NO:3;
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- b) detecting binding of the tetramers to the PBLs wherein the binding of the tetramers to PBLs is indicative of the presence of MTB infection or exposure in the subject.
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A method for determining efficacy of a drug for treatment or prevention of infection or latency of MTB infection in a human subject, comprising:
- a) obtaining a sample comprising PBLs from a patient with a DR4 allelle undergoing treatment for latent MTB using the drug;
- b) contacting the sample under suitable binding conditions with a solid support having bound tetramers of HLA monomers or modified monomers selected from at least one of the following:
- 1) HLA-DR4 monomers or modified monomers, having a bound HLA-binding peptide comprising an amino acid sequence selected from the amino acid sequences set forth in SEQ ID NO:
- 6, 7, 8;
- and
- 2) HLA-A2 monomers or modified monomers, having a bound HLA-binding peptide comprising an amino acid sequence as set forth in SEQ ID NO:
- 1, 2 or 3;
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- c) detecting the amount of binding of the tetramers to the PBLs;
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- d) repeating a), b) and c) after a suitable interval of treatment time,
- wherein a decrease in the amount of the binding after the suitable interval of treatment time indicates efficacy of the drug and a lack of decrease indicates lack of efficacy of the drug for treatment or prevention of infection or latency of MTB in the human.
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A method comprising:
- a) contacting PBLs obtained from a subject with tetramers of HLA monomers or modified monomers, having a bound HLA-binding peptide,
- wherein the subject has an HLA-DR4 allelle and the binding peptide comprises an amino acid sequence as set forth in SEQ ID NO:8, or wherein the subject has an HLA-A2 allelle and the binding peptide comprises an amino acid sequence as set forth in SEQ ID NO:3;
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- b) detecting binding of the tetramers to the PBLs to identify the presence of M. leprae or M. tuberculosis infection or exposure in the subject.
An isolated human HLA-A2-restricted T-cell epitope derived from Mycobacterium tuberculosis (MTB), said epitope comprising an amino acid sequence selected from the group consisting of SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, and conservative variations thereof.
A method for diagnosing the presence of an infection by or exposure to MTB in a human subject, comprising:
- a) contacting PBLs obtained of the subject with tetramers of HLA-A2 monomers, or modified monomers, oligomer or multimers having a bound HLA-binding peptide comprising an amino acid sequence as set forth in any of SEQ ID NO:21-25;
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- b) detecting binding of the tetramers to the PBLs wherein the binding of the tetramers to PBLs is indicative of the presence of MTB infection or exposure in the subject.
A method for determining efficacy of a drug for treatment or prevention of infection or latency of MTB infection in a human subject, comprising:
- a) obtaining a sample comprising PBLs from a patient with a DR4 allelle undergoing treatment for latent MTB using the drug;
- b) contacting the sample under suitable binding conditions with a solid support having bound tetramers of HLA monomers or modified monomers selected from at least one of the following:
- 1) HLA-DR4 monomers or modified monomers, having a bound HLA-binding peptide comprising an amino acid sequence selected from the amino acid sequences set forth in SEQ ID NO:
- 6, 7, 8;
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- 2) HLA-A2 monomers or modified monomers, having a bound HLA-binding peptide comprising an amino acid sequence as set forth in SEQ ID NO:
- 1, 2 ,3, 21, 22, 23, 24, or 25;
- and
Dependencies are inferred from the claim text.