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
- 92Citations
- 20Captures
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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
http://www.sciencedirect.com/science/article/pii/S0006497120710517; http://dx.doi.org/10.1182/blood.v94.9.3094; http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=0033229704&origin=inward; https://ashpublications.org/blood/article/94/9/3094/252472/Rapid-Determination-of-EpsteinBarr-VirusSpecific; https://dx.doi.org/10.1182/blood.v94.9.3094
American Society of Hematology
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