Detection of interleukin-22/17-producing lymphocytes subpopulations in peripheral blood of human by flow cytometry and application in patients with tuberculosis

doi:10.3969/j.issn.1673-8640.2013.01.015

Abstract

Abstract: Objective　To establish a method of detecting different interleukin 22（IL-22）producing lymphocyte subpopulations in human peripheral blood by flow cytometry（FCM)，and investigate its application in patients with tuberculosis（TB）and realize the role of IL-22 producing lymphocyte subpopulations in anti-infection immunity against TB. Methods　Peripheral blood samples collected from 22 TB patients and 18 healthy controls were mixed with RPMI-1640 culture medium at the scale of the same volume. After adding phorbol ester（PMA）and calcium ionomycin mixed for 2 h culturing on the conditions of 37℃ and 5% CO₂，monensin was added and continued being cultured for 4 h. The cells were collected and stained surface molecular or intracellular with fluorochrome-conjugated monoclonal antibodies. The respective proportions of different IL-22 and IL-17 producing lymphocyte subpopulations were detected by FCM. Results　The condition that the concentration of PMA was 100ng/mL and that of ionomycin was 1μg/mL with 2 h being stimulated and cultured could produce IL-22 effectively. The proportions of IL-22 producing gamma delta T cells in TB patients（7.37%）were obviously higher than those in healthy controls（1.81%，P＜0.05). No statistical significance was found between the proportions of IL-22 producing positive cells in CD4^＋T，CD8^＋T，natural killer（NK）cells and B cells in TB patients and those in healthy controls （P＞0.05). No statistical significance was found between the proportions of Th22 and Th17 in TB patients and those in healthy controls （P＞0.05). Conclusions　The established easy，complete and reliable method for the detection of IL-22 producing lymphocyte subpopulations and IL-22 producing gamma delta T cells could act immunological effects in anti-infection immunity against TB.

Key words: Interleukin 22, Interleukin 17, Lymphocyte, Tuberculosis, Flow cytometry

LIANG Hua 1，LIANG Youbao 2，HE Wenxin 3，YANG Junping 1，SUN Guohong 1，YANG Xiaojun 1，LI Baiqing 3. Detection of interleukin-22/17-producing lymphocytes subpopulations in peripheral blood of human by flow cytometry and application in patients with tuberculosis[J]. , 2013, 28(1): 61-66.

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