流式微球分析法检测血清中Th1/Th2/Th17型细胞因子的表达在肺结核病患者中的临床意义
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摘要
目的探讨流式微球分析法(CBA法)检测血清Th1/Th2/Th17型细胞因子的表达在结核病患者中的临床意义。方法按照BD CBA Flex Set检测试剂盒的要求,用CBA法检测98例肺结核患者(痰涂片阳性肺结核患者38例,痰涂片阴性肺结核患者60例)和79例同期健康体检者的Th1型细胞因子(白细胞介素2、γ干扰素、肿瘤坏死因子),Th2型细胞因子(白细胞介素4、白细胞介素6、白细胞介素10)以及Th17型细胞因子(白细胞介素17A)的表达水平,并分析3组之间的差异。结果除白细胞介素17A组间、组内差异无统计学意义(P>0.05)外,结核组白细胞介素2、白细胞介素4、白细胞介素6、白细胞介素10、肿瘤坏死因子、γ干扰素水平均高于对照组(P<0.05);痰涂片阳性患者白细胞介素2、白细胞介素4、白细胞介素10、γ干扰素水平均高于对照组(P<0.05),痰涂片阳性患者和阴性患者差异无统计学意义(P>0.05);而痰涂片阳性患者白细胞介素6和肿瘤坏死因子水平均显著高于痰涂片阴性患者(P<0.05)。结论白细胞介素6和肿瘤坏死因子可作为判断结核病严重程度的指标,监测结核病病情的转归。与此同时,CBA法作为1种新兴的检测技术,敏感度高,可同时检测多种细胞因子,并能大大节约血清用量与时间,因此可作为结核病检测和病情监测的实验室技术。
Objective To evaluate the significance of cytometric beads array(CBA)in the detection of Th1/Th2/Th17 cytokine in patients with pulmonary tuberculosis.Methods The levels of 7cytokines(IL-2,IL-4,IL-6,IL-10,TNF,IFN-γand IL-17A)were detected by CBA according to the instruction of BD CBA Human Th1/Th2/Th17 Cytokine Kit(BD Biosciences,USA)in the serum of 98 patients with tuberculosis(38sputum-smear positive and 60sputum-smear negative)and 79 healthy individuals.Then analyze the relationship and differences among these groups.Results Except IL-17 A,the level of IL-2,IL-4,IL-6,IL-10,TNF,IFN-γwas higher in the patients group than control group;As to IL-2,IL-4,IL-10 and IFN-γ,the sputum-smear positive group had a higher level than control group,but the sputum-smear positive group had no difference with sputum-smear negative group.Compared with sputum-smear negative group,the sputum-smear positive group have a higher level of IL-6and TNF.Conclusion IL-6and TNF can be used to monitor the prognosis of tuberculosis.Meanwhile,CBA is sensitive,could detect several cytokines once with less sample consumption and time,thus could be applied for the monitor of tuberculosis progress.
Objective To evaluate the significance of cytometric beads array(CBA)in the detection of Th1/Th2/Th17 cytokine in patients with pulmonary tuberculosis.Methods The levels of 7cytokines(IL-2,IL-4,IL-6,IL-10,TNF,IFN-γand IL-17A)were detected by CBA according to the instruction of BD CBA Human Th1/Th2/Th17 Cytokine Kit(BD Biosciences,USA)in the serum of 98 patients with tuberculosis(38sputum-smear positive and 60sputum-smear negative)and 79 healthy individuals.Then analyze the relationship and differences among these groups.Results Except IL-17 A,the level of IL-2,IL-4,IL-6,IL-10,TNF,IFN-γwas higher in the patients group than control group;As to IL-2,IL-4,IL-10 and IFN-γ,the sputum-smear positive group had a higher level than control group,but the sputum-smear positive group had no difference with sputum-smear negative group.Compared with sputum-smear negative group,the sputum-smear positive group have a higher level of IL-6and TNF.Conclusion IL-6and TNF can be used to monitor the prognosis of tuberculosis.Meanwhile,CBA is sensitive,could detect several cytokines once with less sample consumption and time,thus could be applied for the monitor of tuberculosis progress.
引文
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[15]王琳,蔡映云,程秋兰,等.肺结核患者的Th1/Th2细胞因子失衡[J].中华结核和呼吸杂志,2002,25(9):535-537.
[16]李红,唐神结.肺结核患者外周血sIL-2R、TNF-α、IFN-γ、IL-6的检测及意义[J].中国防痨杂志,2011,33(1):57-60.
[17]沈云飞,殷凯生,王新宁.肺结核与肺癌患者血IL-12、IFN-γ和IL-4的联合检测及意义[J].江苏医药,2006,32(12):1110-1111.
[2]张培元.肺结核诊断和治疗指南[J].中华结核和呼吸杂志,2001,24(2):70-74.
[3]Winslow GM,Cooper A,Reiley W,et al.Early T-cell responses in tuberculosis immunity[J].Immunol Rev,2008,225(1):284-299.
[4]邓国防,雷建平.结核病与相关免疫细胞和细胞因子[J].中国防痨杂志,2008,30(5):456-460.
[5]Sutherland JS,Young JM,Peterson KL,et al.Polyfunctional CD4+and CD8+T cell responses to tuberculosis antigens in HIV-1-infected patients before and after antiretroviral treatment[J].J Immunol,2010,184(11):6537-6544.
[6]Zumla A,George A,Sharma V,et al.The WHO 2014global tuberculosis report-further to go[J].The Lancet Global Health,2015,3(1):e10-12.
[7]Lasco TM,Yamamoto T,Yoshimura T,et al.Effect of mycobacterium bovis BCG vaccination on mycobacteriumspecific cellular proliferation and tumor necrosis factor alpha production from distinct Guinea pig leukocyte populations[J].Infect Immun,2003,71(12):7035-7042.
[8]Harris J,De Haro SA,Master SS,et al.T helper 2cytokines inhibit autophagic control of intracellular Mycobacterium tuberculosis[J].Immunity,2007,27(3):505-517.
[9]Leal IS,Smedegard B,Andersen P,et al.Interleukin-6and interleukin-12participate in induction of a type 1protective T-cell response during vaccination with a tuberculosis subunit vaccine[J].Infect Immun,1999,67(11):5747-5754.
[10]De La Barrera S,Aleman M,Musella R,et al.IL-10down-regulates costimulatory molecules on Mycobacterium tuberculosis-pulsed macrophages and impairs the lytic activity of CD4and CD8 CTL in tuberculosis patients[J].Clin Exp Immunol,2004,138(1):128-138.
[11]Basu R,Hatton RD,Weaver CT.The Th17family:flexibility follows function[J].Immunol Rev,2013,252(1):89-103.
[12]朱晓燕,陈慧,徐俊驰,等.肺结核患者外周血sIL-2R、IL-6、IL-8、IL-10及TNF-α的检测意义[J].临床肺科杂志,2015(9):1564-1566.
[13]南德齐.肺结核患者血清TNF和IL-6联检的临床意义[J].放射免疫学杂志,2000,13(6):364.
[14]唐神结,肖和平,范以虎,等.肺结核患者血清前炎细胞因子及其受体的变化[J].中华结核和呼吸杂志,2002,25(6):325-329.
[15]王琳,蔡映云,程秋兰,等.肺结核患者的Th1/Th2细胞因子失衡[J].中华结核和呼吸杂志,2002,25(9):535-537.
[16]李红,唐神结.肺结核患者外周血sIL-2R、TNF-α、IFN-γ、IL-6的检测及意义[J].中国防痨杂志,2011,33(1):57-60.
[17]沈云飞,殷凯生,王新宁.肺结核与肺癌患者血IL-12、IFN-γ和IL-4的联合检测及意义[J].江苏医药,2006,32(12):1110-1111.