• Open Access

Conditional ligands for Asian HLA variants facilitate the definition of CD8+ T-cell responses in acute and chronic viral diseases

Authors

  • Cynthia X. L. Chang,

    1. Department of Microbiology, National University of Singapore, Singapore
    2. Immunology Programme, National University of Singapore, Singapore
    3. NUS Graduate School for Integrative Sciences and Engineering, National University of Singapore, Singapore
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  • Anthony T. Tan,

    1. Infection and Immunity Programme, Singapore Institute for Clinical Sciences, Agency for Science, Technology and Research, Singapore
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  • Ming Yan Or,

    1. Department of Microbiology, National University of Singapore, Singapore
    2. Immunology Programme, National University of Singapore, Singapore
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  • Kai Yee Toh,

    1. Department of Microbiology, National University of Singapore, Singapore
    2. Immunology Programme, National University of Singapore, Singapore
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  • Pei Yiing Lim,

    1. Department of Microbiology, National University of Singapore, Singapore
    2. Immunology Programme, National University of Singapore, Singapore
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  • Adeline S. E. Chia,

    1. Infection and Immunity Programme, Singapore Institute for Clinical Sciences, Agency for Science, Technology and Research, Singapore
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  • Thomas M. Froesig,

    1. Center for Cancer Immune Therapy, Department of Hematology, University Hospital Herlev, Herlev, Denmark
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  • Karen D. Nadua,

    1. Program of Emerging Viral Diseases, Duke-NUS Graduate Medical School, National University of Singapore, Singapore
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  • Hsueh-Ling J. Oh,

    1. Institute of Molecular and Cell Biology, Agency for Science, Technology and Research, Singapore
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  • Hoe Nam Leong,

    1. Singapore General Hospital, Singapore
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  • Sine R. Hadrup,

    1. Center for Cancer Immune Therapy, Department of Hematology, University Hospital Herlev, Herlev, Denmark
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  • Adam J. Gehring,

    1. Infection and Immunity Programme, Singapore Institute for Clinical Sciences, Agency for Science, Technology and Research, Singapore
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  • Yee-Joo Tan,

    1. Department of Microbiology, National University of Singapore, Singapore
    2. Institute of Molecular and Cell Biology, Agency for Science, Technology and Research, Singapore
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  • Antonio Bertoletti,

    1. Department of Medicine, National University of Singapore, Singapore
    2. Infection and Immunity Programme, Singapore Institute for Clinical Sciences, Agency for Science, Technology and Research, Singapore
    3. Program of Emerging Viral Diseases, Duke-NUS Graduate Medical School, National University of Singapore, Singapore
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  • Gijsbert M. Grotenbreg

    Corresponding author
    1. Department of Biological Sciences, National University of Singapore, Singapore
    2. Immunology Programme, National University of Singapore, Singapore
    3. NUS Graduate School for Integrative Sciences and Engineering, National University of Singapore, Singapore
    • Department of Microbiology, National University of Singapore, Singapore
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Full Correspondence: Dr. Gijsbert Grotenbreg, Immunology Programme, Life Sciences Institute #03-05, 28 Medical Drive, Singapore 117456

Fax: +65-6778-2684

e-mail: grotenbreg@nus.edu.sg

Abstract

Conditional ligands have enabled the high-throughput production of human leukocyte antigen (HLA) libraries that present defined peptides. Immunomonitoring platforms typically concentrate on restriction elements associated with European ancestry, and such tools are scarce for Asian HLA variants. We report 30 novel irradiation-sensitive ligands, specifically targeting South East Asian populations, which provide 93, 63, and 79% coverage for HLA-A, -B, and -C, respectively. Unique ligands for all 16 HLA types were constructed to provide the desired soluble HLA product in sufficient yield. Peptide exchange was accomplished for all variants as demonstrated by an ELISA-based MHC stability assay. HLA tetramers with redirected specificity could detect antigen-specific CD8+ T-cell responses against human cytomegalovirus, hepatitis B (HBV), dengue virus (DENV), and Epstein-Barr virus (EBV) infections. The potential of this population-centric HLA library was demonstrated with the characterization of seven novel T-cell epitopes from severe acute respiratory syndrome coronavirus, HBV, and DENV. Posthoc analysis revealed that the majority of responses would be more readily identified by our unbiased discovery approach than through the application of state-of-the-art epitope prediction. This flow cytometry-based technology therefore holds considerable promise for monitoring clinically relevant antigen-specific T-cell responses in populations of distinct ethnicity.

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