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BACKGROUND: Clinical investigation of antigen-specific T cells in potentially immunodeficient patients is an important and often challenging aspect of patient diagnostic work up. Methods for detection of microbial exposure to the T-cell compartment exist but are laborious and time consuming. Recently, a whole blood technique involving flow cytometry and detection of CD25 and OX40 (CD134) expression on the surface of activated CD4+ T cells was shown to be accurate and concordant when compared with more traditional methods of antigen-specific T-cell detection. METHODS: Whole heparinized blood was collected from healthy donors and set up using the "OX40" assay to detect antigen-specific CD4+ T-cell responses to Varicella Zoster Virus, Epstein-Barr Virus (EBV), Cytomegalovirus, Candida albicans, and Streptococcus pneumoniae. RESULTS: The "OX40" assay technique was clinically validated for routine use in an NHS clinical immunology laboratory by analysis of incubation length (40-50 h), sample transport time (up to 24 h at room temperature), concordance with serology testing, proliferation and interferon-gamma production. In addition, 63 healthy controls (age range 21-78) were tested for responses to generate a healthy control reference range. CONCLUSIONS: The OX40 assay, as presented in this report, represents an economical, rapid, robust whole blood technique to detect antigen-specific T cells, which is suitable for clinical immunology diagnostic laboratory use.

Original publication




Journal article


Cytometry B Clin Cytom

Publication Date





350 - 361


CD4+ T cells, costimulation/costimulatory, diagnostics, flow cytometry, Adult, Aged, CD4-Positive T-Lymphocytes, Candida albicans, Cytomegalovirus, Female, Flow Cytometry, Herpesvirus 3, Human, Herpesvirus 4, Human, Humans, Interferon-gamma, Interleukin-2 Receptor alpha Subunit, Male, Middle Aged, Receptors, OX40, Reference Values, Streptococcus pneumoniae, Young Adult