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Comparative analysis of whole‐blood interferon‐γ and flow cytometry assays for detecting post‐treatment Immune responses in patients with active tuberculosis
Author(s) -
Kim Chang Ho,
Choi Keum Ju,
Yoo Seung Soo,
Lee Shin Yup,
Won Dong Il,
Lim Jeong Ok,
Cha Seung Ick,
Park Jae Yong,
Lee Jaehee
Publication year - 2014
Publication title -
cytometry part b: clinical cytometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.646
H-Index - 61
eISSN - 1552-4957
pISSN - 1552-4949
DOI - 10.1002/cyto.b.21110
Subject(s) - tuberculosis , medicine , antigen , immunology , mycobacterium tuberculosis , immune system , flow cytometry , interferon gamma , cytokine , t cell , interferon , tumor necrosis factor alpha , pathology
Background Intracellular cytokine flow cytometry (ICCFC) has been explored to detect tuberculosis (TB) infections; however, there are little data regarding its use to examine the dynamic responses of Mycobacterium tuberculosis (MTB)‐specific T‐cells after anti‐tuberculous therapy. The aim of this study was to analyze both dynamic changes in functional MTB antigen‐specific T‐cell subsets and interferon‐gamma (IFN‐γ) levels using ICCFC and the QuantiFERON‐TB Gold In‐Tube (QFT‐IT) test, respectively, following anti‐tuberculous treatment in patients with active TB. Methods Twenty‐six patients with active TB were enrolled in the study, and QFT‐IT and ICCFC were performed simultaneously both before and after treatment. IFN‐γ levels (QFT‐IT test) and the numbers of IFN‐γ‐ or tumor necrosis factor‐alpha (TNF‐α)‐expressing T‐cells (ICCFC assay) were examined after stimulation with MTB antigen. Results There was no significant reduction in the mean IFN‐γ concentrations measured by the QFT‐IT test after anti‐tuberculous treatment ( P = 0.314). ICCFC analysis showed that the numbers of IFN‐γ + /CD4 ‐ T‐cells, and CD4 + T‐cells producing TNF‐α, either alone or in combination with IFN‐γ, were significantly reduced after anti‐tuberculous treatment. The IFN‐γ + /TNF‐α + /CD4 + T‐cell subset showed the greatest difference between untreated and treated patients with active TB (area under the curve = 0.734, P = 0.004). Conclusions Unlike the QFT‐IT test, ICCFC provides diverse immunological information about dynamic changes in the number of MTB antigen‐specific T‐cells following anti‐tuberculous therapy. Thus, analysis of MTB antigen‐stimulated T‐cell responses using ICCFC might have a role to play in monitoring treatment responses in patients with active TB. © 2013 International Clinical Cytometry Society