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Identification of Mycobacterium tuberculosis PPE68-Specific HLA-A*0201-Restricted Epitopes for Tuberculosis Diagnosis

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Abstract

PPE68 is a Mycobacterium tuberculosis-specific protein which is absent from the vaccine strains of BCG. A panel of 14 PPE68-derived peptides predicted to bind to HLA-A*0201 was synthesized. The HLA-A*0201 restriction of these peptides was determined in T2 cell line and HLA-A*0201 transgenic mice. The specificity of peptides was assessed in pulmonary tuberculosis (TB) patients using IFN-γ enzyme-linked immunospot (ELISPOT) assay, and immunodominant peptides were further used to evaluate their diagnostic potential in HLA-A*0201-positive pulmonary TB patients. 13 out of 14 peptides were identified as high-affinity binders. Of these peptides, 12 peptides induced significant IFN-γ-secreting T cell response in transgenic mice and 9 peptides were efficiently recognized by peripheral blood mononuclear cells of 10 HLA-A*0201-positive TB patients. Four immunodominant HLA-A*0201-restricted epitopes (PPE68126–134, PPE68133–141, PPE68140–148, and PPE68148–156) were recognized by the most of 80 HLA-A*0201-positive TB patients (81, 86, 74, and 84 %, respectively). These epitopes may be used for a potential diagnosis of M. tuberculosis infection.

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Acknowledgments

The work was funded by the National Natural Science Foundation of China (30972763, 81261160323, 81172811, 81171576 and 31070143), National Science and Technology Key Projects for Major Infectious Diseases (2013ZX10003001), the Natural Science Foundation of Zhejiang (LY13H160035 and LQ14C010006), the Planned Science and Technology Project of Ningbo (2012A610247), and the Planned Science and Technology Project of Zhejiang (2014C33261).

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    Authors and Affiliations

    1. Institute of Arboviruses, School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou, China

      Zhi-liang Duan, Qiang Li, Sina Wang, Hui-fang Liu, Bo-kun Chen, Xi Huang & Jin-sheng Wen

    2. Department of Clinical Laboratory, The Second Affiliated Hospital & Yuying Children’s Hospital of Wenzhou Medical University, Wenzhou, China

      Zhi-liang Duan & Xin-yu Chen

    3. Department of Clinical Laboratory, The Third Affiliated Hospital of Wenzhou Medical University, Wenzhou, China

      Qiang Li

    4. Department of Immunology, Institute of Tuberculosis Control, Institute of Human Virology, Sun Yat-sen University Zhongshan School of Medicine, Guangzhou, China

      Xi Huang

    5. Key Laboratory of Tropical Diseases Control (Sun Yat-sen University), Ministry of Education, Guangzhou, China

      Xi Huang

    6. Department of Liver, The Secondary Hospital of Ningbo, Ningbo, China

      De-zhou Li

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    1. Zhi-liang Duan
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    Correspondence to Xi Huang or Jin-sheng Wen.

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    Zhi-liang Duan and Qiang Li have contributed equally to this work.

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    Duan, Zl., Li, Q., Wang, S. et al. Identification of Mycobacterium tuberculosis PPE68-Specific HLA-A*0201-Restricted Epitopes for Tuberculosis Diagnosis. Curr Microbiol 70, 769–778 (2015). https://doi.org/10.1007/s00284-015-0786-x

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    • DOI: https://doi.org/10.1007/s00284-015-0786-x

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