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Rapid Determination of Epstein-Barr Virus–Specific CD8 + T-Cell Frequencies by Flow Cytometry

Blood, ISSN: 0006-4971, Vol: 94, Issue: 9, Page: 3094-3100
1999
  • 92
    Citations
  • 0
    Usage
  • 20
    Captures
  • 0
    Mentions
  • 0
    Social Media
Metric Options:   Counts1 Year3 Year

Metrics Details

  • Citations
    92
    • Citation Indexes
      92
  • Captures
    20

Article Description

We have developed an efficient and rapid method for detection of Epstein-Barr virus (EBV)-specific CD8 + T-cell frequencies both in freshly isolated peripheral blood mononuclear cells (PBMCs) and in vitro established cytotoxic T lymphocyte (CTL) lines. Responder cells are thereby stimulated with an autologous lymphoblastoid cell line for 5 hours and intracellular accumulation of interferon γ (IFNγ) is detected by multiparameter flow cytometric analysis. EBV-specific CD8 + T-cell frequencies ranged between 0.63% and 1.29% in PBMCs of 5 healthy long-term EBV carriers. Using EBV-specific T-cell lines, it was shown that flow cytometric analysis is more sensitive than limiting dilution analysis for CTL precursors and enzyme-linked immunospot assay detecting IFNγ-producing T cells. The class I restriction of IFNγ production was confirmed using an anti-class I monoclonal antibody (MoAb). Information on other cytokine production of EBV-specific CTLs could be obtained using combinations of anti-cytokine MoAbs. The sensitive and rapid nature of the flow cytometric assay for EBV-specific CD8 + T-cell frequency has significant advantages for evaluation of EBV-specific CD8 + T-cell responses in PBMCs of patients with EBV-related diseases.

Bibliographic Details

Kiyotaka Kuzushima; Yo Hoshino; Ken Fujii; Naoaki Yokoyama; Masatoshi Fujita; Tohru Kiyono; Hiroshi Kimura; Tsuneo Morishima; Yasuo Morishima; Tatsuya Tsurumi

American Society of Hematology

Biochemistry, Genetics and Molecular Biology; Immunology and Microbiology; Medicine

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