流式细胞术检测人外周血产生IL-22和/或IL-17的淋巴细胞亚群及其在结核病患者中的应用

doi:10.3969/j.issn.1673-8640.2013.01.015

检验医学 ›› 2013, Vol. 28 ›› Issue (1): 61-66.DOI: 10.3969/j.issn.1673-8640.2013.01.015

• 技术研究与评价.论著 • 上一篇下一篇

流式细胞术检测人外周血产生IL-22和/或IL-17的淋巴细胞亚群及其在结核病患者中的应用

梁华¹,梁友宝²,贺文欣³,杨军平¹,孙国宏¹,杨小军¹,李柏青⁴

1. 江西中医学院附属医院检验科，江西南昌 330006；2.蚌埠市第一医院检验科，安徽蚌埠 233030；3.蚌埠医学院免疫学教研室安徽省感染与免疫重点实验室，安徽蚌埠 233030

收稿日期:2012-11-26 修回日期:2012-08-24 出版日期:2013-01-30 发布日期:2013-01-10
通讯作者: 李柏青，联系电话：0552-3171086。
作者简介:梁华，男，1975年生，硕士，主管技师，主要从事细胞免疫学研究。
基金资助:
国家科技重大专项基金资助项目(2008ZX10003-011)

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

1. Department of Clinical Laboratory，the Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine，Jiangxi Nanchang 330006，China；2. Department of Clinical Laboratory，the First Hospital of Bengbu，Anhui Bengbu 233030，China；3. Department of Immunology，Bengbu Medical College，Anhui Key Laboratory of Infection and Immunity，Anhui Bengbu 233030，China

Received:2012-11-26 Revised:2012-08-24 Online:2013-01-30 Published:2013-01-10

摘要/Abstract

摘要： 目的　建立用流式细胞术(FCM)检测人外周血产白细胞介素22(IL-22)的不同淋巴细胞亚群的方法，并探讨其在结核病(TB)患者中的应用，以了解产IL-22的淋巴细胞亚群在抗结核感染免疫中的作用。方法　采集22例TB患者和18名健康对照者外周血，与相同体积RPMI-1640培养液混匀，加入佛波醇酯(PMA)和钙离子霉素，混匀后于37 ℃ 5% CO₂静置培养2 h，再加入莫能霉素后继续培养4 h，收集细胞，用荧光素标记进行细胞表面抗原和胞内细胞因子染色，用FCM检测表达IL-22、IL-17的不同淋巴细胞亚群比例。结果　PMA浓度为100 ng/mL，钙离子霉素浓度为1 μg/mL，刺激培养2 h可最有效刺激IL-22的产生；TB患者外周血中产IL-22的γδT细胞比例（7.37%）明显高于正常人（1.81%，P＜0.05），而在CD4^＋T、CD8^＋T、自然杀伤(NK)细胞和B细胞等其他细胞亚群中，产IL-22的阳性细胞比例在TB患者和健康对照者之间差异无统计学意义（P＞0.05）；Th22和Th17细胞比例在TB患者和健康对照者之间差异无统计学意义（P＞0.05)。结论　建立了一套简便、完整、可靠的FCM检测产IL-22的淋巴细胞亚群的方法，产IL-22的γδT细胞在抗结核感染免疫中可能发挥免疫效应。

关键词: 白细胞介素22, 白细胞介素17, 淋巴细胞, 结核病, 流式细胞术

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

梁华 1，梁友宝 2，贺文欣 3，杨军平 1，孙国宏 1，杨小军 1，李柏青 3. 流式细胞术检测人外周血产生IL-22和/或IL-17的淋巴细胞亚群及其在结核病患者中的应用[J]. 检验医学, 2013, 28(1): 61-66.

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.

参考文献

[1]Dumoutier L，Louahed J，Renauld JC. Cloning and characterization of IL-10-related T cell-derived inducible factor(IL-TIF)，a novel cytokine structurally related to IL-20 and inducible by IL-9[J]. J Immunol，2000，164(4)：1814-1819.
[2]Wolk K，Kunz S，Witte E. IL-22 increases the innate immunity of tissues[J]. Immunity，2004，21(2)：241-254.
[3]Ikeuchi H，Kuroiwa T，Hiramatsu N，et al. Expression of interleukin-22 in rheumatoid arthritis：potential role as a proinflammatory cytokine[J]. Arthritis Rheum，2005，52(4)：1037-1046.
[4]Brand S，Beigel F，Olszak T，et al. IL-22 is increased in active Crohn′s disease and promotes pro-inflammatory gene expression and intestinal epithelial cell migration[J]. Am J Physiol Gastrointest Liver Physiol，2006，290(4)：G827-G838.
[5]Andoh A，Zhang Z，Inatomi O，et al. Interleukin-22，a member of the IL-10 subfamily，induces inflammatory responses in colonic subepithelial myofibroblasts[J]. Gastroenterology，2005，129(3)：969-984.
[6]Eyerich S，Eyerich K，Pennino D，et al. Th22 cells represent a distinct human T cell subset involved in epidermal immunity and remodeling[J]. J Clin Invest，2009，119(12)：3573-3585.
[7]Mosmann TR，Coffman RL. Th1 and Th2 cells：different patterns of lymphokine secretion lead to different functional properties[J].Annu Rev Immunol，1989，7：145-173.
[8]〖JP4〗Harrington LE，Hatton RD，Mangan PR，et al. Interleukin 17-producing CD4 effector T cells develop via a lineage distinct from the T helper type 1 and 2 lineages[J]. Nat Immunol，2005，6(11)：1123-1132.
[9]Korn T，Bettelli E，Oukka M，et al. IL-17 and Th17 cells[J]. Annu Rev Immunol，2009，27：485-517.
[10]庄园，石云，邹全明. IL-22：Th17细胞的重要效应分子[J]. 中国免疫学杂志，2009，25(8)：761-764.
[11]周茂华，吴长有. 不同刺激剂和培养环境对CD4 、CD8 T细胞内细胞因子表达的影响[J]. 细胞与分子免疫学杂志，2007，23(2)：168 -171.
[12]Kemp K，Bruunsqaard H. Identification of IFN-gamma-producing CD4 T cells following PMA stimulation[J]. J Interferon Cytokine Res，2001，21(7)：503-506.
[13]Ordway DJ，Costa L，Martins M，et al. Increased interleukin-4 production by CD8 and gamma delta T cells in health-care workers is associated with the subsequent development of active tuberculosis[J]. J Infect Dis，2004，190(4)：756-766.
[14]Zheng Y，Danlienko DM，Valdez P，et al. Interleukin-22，a T(H）17 cytokine ，mediates IL-23-induced dermal inflammation and acanthosis[J]. Nature，2007，445(7128)：648-651.
[15]Yao S，Huang D，Chen CY，et al.Differentiation，distribution and gamma delta T cell-driven regulation of IL-22-producing T cells in tuberculosis[J]. PLoS Pathog，2010，6(2)：e1000789.
[16]Scriba TJ，Kalsdorf B，Abrahams DA，et al. Distinct，specific IL-17-and IL-22-producing CD4 T cell subsets contribute to the human anti-mycobacterial immune response[J]. J Immunol，2008，180(3)：1962-1970.

流式细胞术检测人外周血产生IL-22和/或IL-17的淋巴细胞亚群及其在结核病患者中的应用

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

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

[1]	楚野, 熊春翔, 黄宇, 曾佳兴, 易波德, 黄能干, 杨屹峰, 杨枫. 基于CAR、NLR的列线图模型在预测老年髋部骨折患者术后1年内死亡中的价值[J]. 检验医学, 2025, 40(9): 834-840.
[2]	王秋菊, 肖辉建, 吴双, 赵婉婷, 代绪波, 庄岳鹏. 外周血淋巴细胞形态学分类在危重型流行性感冒疗效判断中的价值[J]. 检验医学, 2025, 40(9): 901-906.
[3]	赵君, 王鹏. 基于血常规指标和NLR、PLR、MLR诊断儿童腺病毒感染[J]. 检验医学, 2025, 40(8): 817-820.
[4]	王丽, 唐玲, 金亚雄, 张君龙, 高雪丹, 牛倩. LDH、MO#和LMR在RA-ILD鉴别诊断中的价值[J]. 检验医学, 2025, 40(6): 525-533.
[5]	施方静, 胡姗姗, 罗信国, 朱艳, 俞方泉, 叶虹位. 慢性淋巴细胞白血病患者程序性死亡分子1/程序性死亡-配体1的表达及其临床意义[J]. 检验医学, 2025, 40(6): 592-595.
[6]	李亚周, 苏敏, 张景皓, 方毅, 王粟, 王诗雯, 赵虎. CRP、HBP、CLR、HAR单项和联合检测在老年患者血流感染诊断中的价值[J]. 检验医学, 2025, 40(5): 489-492.
[7]	王蜀平, 张文, 王溦雅, 马丽娟. 外周血淋巴细胞亚群和细胞因子在鉴别儿童病毒性脑炎与自身免疫性脑炎中的价值[J]. 检验医学, 2025, 40(3): 223-229.
[8]	郑莹, 陆喆, 薛静. SLE患儿血清25（OH）D3水平与淋巴细胞亚群的关系[J]. 检验医学, 2025, 40(3): 230-234.
[9]	杨含草, 梁可青, 吴茗, 徐锦. 血液免疫学指标在评估自身免疫性脑炎患儿预后中的价值[J]. 检验医学, 2025, 40(3): 235-240.
[10]	高子康, 陈晶. 基于外周血淋巴细胞亚群特征构建抑郁障碍患者预后列线图预测模型[J]. 检验医学, 2025, 40(12): 1146-1152.
[11]	谭阳阳, 杨微, 沈化清, 李碧君, 陆秋霞, 刘熙君, 赖媛媛. 有核红细胞假性升高1例报道[J]. 检验医学, 2025, 40(11): 1132-1134.
[12]	邹珍洋, 于瑾舒, 何仁栋, 谢宁, 郭斌. GDF-15、NT-proBNP和NLR在慢性心力衰竭诊断、病情严重程度和预后评估中的价值[J]. 检验医学, 2025, 40(10): 946-953.
[13]	王梦茹, 周麟, 邢茜茜, 崔倩倩. 老年髋部骨折患者术前NLR、UA和PGE2联合检测对术后谵妄的预测价值[J]. 检验医学, 2025, 40(10): 987-991.
[14]	刘梦娜, 王洪玲, 白萍, 蔡宇, 李莉. ETP-ALL实验室诊断和治疗新进展[J]. 检验医学, 2025, 40(1): 1-7.
[15]	王洪玲, 蔡宇, 刘梦娜, 白萍. POX阳性率>3%的ETP-ALL患者临床特征分析[J]. 检验医学, 2025, 40(1): 20-24.