MTB分泌蛋白抗原85B-早期分泌靶抗原6融合蛋白对结核性胸膜炎的辅助诊断价值

doi:10.3969/j.issn.1000-6621.2018.11.004

摘要/Abstract

摘要：

目的研究MTB分泌蛋白抗原85B(Ag85B)-早期分泌靶抗原6(ESAT6)融合蛋白在结核性胸膜炎辅助诊断中的价值。方法研究对象为2014年1月至2015年1月于河北省胸科医院住院的结核性胸膜炎(结核组)患者45例,非结核性胸膜炎(非结核组)患者26例,以及体检健康人群(健康组)25名。应用流式细胞仪分别检测结核分枝杆菌Ag85B-ESAT6融合蛋白刺激前后结核组、非结核组外周血、胸腔积液及健康组外周血中γ干扰素(IFN-γ)的含量,采用免疫组织化学染色法检测结核组与非结核组患者组织(通过手术或穿刺活检获得的胸膜组织)中CD4 ⁺和CD8 ⁺细胞的表达水平。采用SPSS 13.0软件进行统计学处理,数据符合正态分布的组间比较采用t检验,以P<0.05为差异有统计学意义。结果 MTB Ag85B-ESAT6融合蛋白刺激前后,结核组外周血中IFN-γ的含量分别为(42.63±10.51)pg/ml和(401.90±72.54)pg/ml;非结核组分别为(38.97±7.08)pg/ml和(40.04±6.80)pg/ml;健康组分别为(39.61±7.28)pg/ml和(39.86±6.97)pg/ml。结核组胸腔积液中IFN-γ的含量分别为(411.91±41.56)pg/ml和(1342.67±167.96)pg/ml;非结核组分别为(47.99±11.49)pg/ml和(48.76±11.25)pg/ml。刺激前结核组胸腔积液中IFN-γ的含量明显高于非结核组,两组比较差异有统计学意义(t=55.194,P=0.000);刺激后结核组外周血单个核细胞产生的IFN-γ含量相对于刺激前及刺激后的其他两组比较,差异均有统计学意义(t=32.879、33.211和33.204,P值均为0.000);刺激后结核组与刺激前及非结核组刺激前后胸腔积液中IFN-γ的含量比较,差异均具有统计学意义(t=36.085、51.478和51.499,P值均为0.000)。免疫组织化学染色结果显示结核组与非结核组患者胸膜组织中的CD4 ⁺和CD8 ⁺积累光密度值(结核组:16349.91±2376.36和10525.77±1164.86;非结核组:1853.64±670.40和1327.15±175.55)差异均具有统计学意义(t=14.381和19.127,P值均为0.000)。结论结核分枝杆菌Ag85B-ESAT6融合蛋白刺激外周血和胸腔积液后均明显提高结核特异性IFN-γ的含量,对结核性胸膜炎的辅助诊断有较高的临床应用价值。

关键词: 结核, 胸膜, 干扰素γ, 实验室技术和方法, 评价研究, Ag85B-ESAT6融合蛋白

Abstract:

Objective To explore the value of Mycobacterium tuberculosis antigen 85B (Ag85B) and early secreted antigenic target 6-kDa antigen (ESAT6) fusion protein in the diagnosis of tuberculous pleurisy.Methods Inpatients and healthy controls were enrolled in Hebei Chest Hospital from January 2014 to January 2015, including 45 patients with tuberculous pleurisy, 26 patients with non-tuberculous pleurisy, and 25 cases of healthy controls. Flow cytometry was used to detect the IFN-γ concentrations in the serum, and in the pleural effusion with and without the Ag85B-ESAT6 fusion protein stimulation. The number of CD4 ⁺ and CD8 ⁺cells in tuberculous pleurisy group and non-tuberculous pleurisy group patients was detected by immunohistochemical staining. The statistic analysis was performed by SPSS 13.0 software by t test, P<0.05 was considered as statistically significant.Results With and without stimulation of Mycobacterium tuberculosis Ag85B-ESAT6 fusion protein, the concentrations of IFN-γ in the blood of tuberculous pleurisy group were (42.63±10.51)pg/ml and (401.90±72.54)pg/ml, respectively, and non-tuberculous pleurisy group were (38.97±7.08)pg/ml and (40.04±6.80)pg/ml, respectively, and healthy control-group were (39.61±7.28)pg/ml and (39.86±6.97)pg/ml, respectively. Under the same condition, the concentrations of IFN-γ in tuberculous pleurisy group pleural effusion was (411.91±41.56)pg/ml and (1342.67±167.96)pg/ml, respectively, and non-tuberculous pleurisy group were (47.99±11.49)pg/ml and (48.76±11.25)pg/ml, respectively. The concentration of pleural effusion IFN-γ without stimulation was significantly higher in tuberculous pleurisy group than that in non-tuberculous pleurisy group(t=55.194, P=0.000). The stimulated concentration of blood IFN-γ in tuberculous pleurisy group was significantly higher than those in the other groups whether stimulated or not(t=32.879, 33.211, 33.204, P=0.000, 0.000, 0.000). The stimulated concentration of pleural effusion IFN-γ was significantly higher than those in the other groups whether stimulated or not(t=36.085, 51.478, 51.499, P=0.000, 0.000, 0.000). Immunohistochemical staining showed that the numbers of CD4 ⁺ and CD8 ⁺ cells in tissues between tuberculous pleurisy group (16349.91±2376.36, 10525.77±1164.86) and non-tuberculous pleurisy group (1853.64±670.40, 1327.15±175.55) was significantly different (t=14.381, 19.127, all P=0.000). Conclusion Mycobacterium tuberculosis Ag85B-ESAT6 fusion protein significantly increased the concentration of IFN-γ after stimulation of peripheral blood and pleural effusion, and showed potential clinical value for the diagnosis of tuberculous pleurisy.

Key words: Tuberculosis, pleural, Interferon-gamma, Laboratory techniques and procedures, Evaluation studies, Ag85B-ESAT6 fusion protein

安晓颖,杨永辉,朱桂云,方鹏,白洪忠,章志华. MTB分泌蛋白抗原85B-早期分泌靶抗原6融合蛋白对结核性胸膜炎的辅助诊断价值[J]. 中国防痨杂志, 2018, 40(11): 1164-1169. doi: 10.3969/j.issn.1000-6621.2018.11.004

AN Xiao-ying,YANG Yong-hui,ZHU Gui-yun,FANG Peng,BAI Hong-zhong,ZHANG Zhi-hua. Diagnostic value of Mycobacterium tuberculosis antigen 85B and early secreted antigenic target 6-kDa antigen fusion protein for tuberculous pleurisy[J]. Chinese Journal of Antituberculosis, 2018, 40(11): 1164-1169. doi: 10.3969/j.issn.1000-6621.2018.11.004

图/表 6

表1

不同组别研究对象的性别和年龄比较

组别	性别(例或名)		χ²值	P值	年龄 (岁, $x ˉ$ ±s)	t值	P值
组别	男	女	χ²值	P值	年龄 (岁, $x ˉ$ ±s)	t值	P值
结核组	31	14	1.607^a	0.205	35.13±6.77	0.960^a	0.340
非结核组	14	12	1.163^b	0.281	33.46±7.56	-0.232^b	0.818
健康组	14	11	0.024^c	0.877	33.96±7.81	0.658^c	0.513

表1

表2

不同组别MTB Ag85B-ESAT6融合蛋白刺激前血清中IFN-γ含量比较

组别	例数(例或名)	IFN-γ(pg/ml, $x ˉ$ ±s)	t值	P值
结核组	45	42.63±10.51	1.747^a	0.085
非结核组	26	38.97±7.08	-0.317^b	0.752
健康组	25	39.61±7.28	1.412^c	0.163

表2

表3

结核与非结核组胸腔积液行MTB Ag85B-ESAT6融合蛋白刺激前后的IFN-γ含量比较

组别	总例数	刺激后IFN-γ(pg/ml, $x ˉ$ ±s)	刺激前IFN-γ(pg/ml, $x ˉ$ ±s)	t值	P值
结核组	45	1342.67±167.96	411.91±41.56	36.085	0.000
非结核组	26	48.76±11.25	47.99±11.49	0.244	0.808

表3

表4

不同组别MTB Ag85B-ESAT6融合蛋白刺激外周血中单个核细胞产生IFN-γ含量比较

组别	例数(例或名)	IFN-γ含量(pg/ml, $x ˉ$ ±s)	t值	P值
结核组	45	401.90±72.54	33.211^a	0.000
非结核组	26	40.04±6.80	0.094^b	0.925
健康组	25	39.86±6.97	33.204^c	0.000

表4

表5

不同组别MTB Ag85B-ESAT6融合蛋白刺激前后外周血中IFN-γ含量的比较

组别	总数(例或名)	刺激后IFN-γ(pg/ml, $x ˉ$ ±s)	刺激前IFN-γ(pg/ml, $x ˉ$ ±s)	t值	P值
结核组	45	401.90±72.54	42.63±10.51	32.879	0.000
非结核组	26	40.04±6.80	38.97±7.08	0.554	0.582
健康组	25	39.86±6.97	39.61±7.28	0.122	0.903

表5

图1

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