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A simple and efficient method of generating HCMV pp65-specific T cells using overlapping peptides.

S-U Hwang, J-H Lee, S-Y Choi

    Acta Virologica
    |November 5, 2020
    PubMed
    Summary

    Overlapping peptides rapidly expand human cytomegalovirus (HCMV)-specific cytotoxic T lymphocytes from peripheral blood mononuclear cells (PBMCs). This method overcomes HLA restriction, generating effective T cells for adoptive cell therapy.

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    Area of Science:

    • Immunology
    • Virology
    • Cell Therapy

    Background:

    • Generating human cytomegalovirus (HCMV)-specific T lymphocytes is challenging due to complex culture methods, long durations, and human leukocyte antigen (HLA) restrictions.
    • Existing methods limit the efficient expansion of cytotoxic T lymphocytes (CTLs) crucial for combating HCMV infections.

    Purpose of the Study:

    • To investigate a novel method for rapidly generating large numbers of HCMV-specific CTLs using overlapping peptides (OLP).
    • To assess the efficacy of OLP stimulation regardless of HLA type for potential adoptive cell therapy.

    Main Methods:

    • Peripheral blood mononuclear cells (PBMCs) from healthy donors were stimulated in vitro with 138 pp65-derived overlapping synthetic peptides.
    • Expansion and specificity of HCMV pp65-specific CD8+ T lymphocytes were analyzed using pentamer staining and interferon-gamma (IFN-γ) expression.
    • Cytolytic activity of generated T lymphocytes against target cells was evaluated.

    Main Results:

    • In vitro stimulation with pp65-derived OLP rapidly expanded HCMV pp65-specific CD8+ T lymphocytes from PBMCs expressing HLA-A*02:01 and HLA-A*24:02.
    • OLP-induced T lymphocytes demonstrated effective cytolytic responses against HCMV-loaded target cells, irrespective of HLA type.
    • Phenotypic analysis confirmed an increase in CD3+ CD8+ T cells without affecting other immune cell populations.

    Conclusions:

    • In vitro stimulation with overlapping peptides is an efficient method for generating substantial numbers of HCMV pp65-specific cytotoxic T lymphocytes.
    • This OLP-based approach overcomes HLA restriction, offering a promising strategy for HCMV-targeted adoptive cell therapy.