Author(s): Won DI, Park JR
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Abstract BACKGROUND: Interferon-gamma (IFN-gamma) release assays and the detection of IFN-gamma synthesis in the cytoplasm of activated CD4+ T cells by flow cytometry have recently been used for tuberculosis (TB) diagnosis. The aim of this study was to compare the performance of IFN-gamma assay between ELISA (QuantiFERON-TB Gold, QFT) and intracellular cytokine flow cytometry (ICCFC), and to investigate the significance of an optimal gating strategy in ICCFC. METHODS: The CD4+ T cell response to TB antigens was measured using the intracellular cytokine staining technique and four color FC (CD3, CD4, IFN-gamma, and tumor necrosis factor-alpha (TNF-alpha)) on whole blood samples. The results were compared with those of QFT. RESULTS: Regarding sensitivity in the TB group, patients in the QFT positive TB group (N = 22) were all ICCFC positive and 9 patients (64\%) in the QFT negative TB group (N = 14) were ICCFC positive. In all test tubes (N = 72), sensitivity of "targeted" gating for TNF-alpha+ IFN-gamma+ CD4+ T cells was significantly higher than customary gating (72\%, 54\%, respectively, P = 0.001). CONCLUSIONS: The diagnostic sensitivity of ICCFC was further confirmed to be much higher than that of QFT. In the ICCFC analysis, TNF-alpha+ IFN-gamma+ CD4+ T cells should be sequentially gated through appropriately defined regions, minimizing interferents and reflecting characteristics of light scatter and marker expressions of CD4+ T cells activated by TB antigens. (c) 2009 Clinical Cytometry Society.
This article was published in Cytometry B Clin Cytom
and referenced in Journal of Infectious Diseases & Preventive Medicine