摘要
目前,抗结核的化疗方案包括2个月的强化期与4个月的巩固期,而治疗过程中特异性细胞免疫应答随细菌载量改变呈现的动态变化特征有可能成为治疗疗效的生物标志物,并为个性化治疗方案提供理论依据。为此,我们收集了9例活动性结核病患者治疗前,治疗2、4、6个月的外周血样本,采用酶联免疫斑点测定法(enzyme-linked immune spot assay,ELISPOT)测定了不同治疗阶段结核特异性抗原ESAT-6和CFP-10刺激下PBMC中分泌IFN-γ的细胞数量,同时分析了抗原特异性Th1型细胞因子分泌水平以及多功能T细胞比例的动态变化特征。结果显示:随着治疗的深入,ESAT-6及CFP-10刺激患者PBMC后,释放IFN-γ的细胞数量显著下降(P<0.05)。CD4^+IFN-γ^+T淋巴细胞的比例在治疗前与治疗6个月时相比有下降趋势,且治疗后结果具有归一性;虽然多功能T细胞有重要抗结核作用,但在我们统计的患者治疗过程中的变化不明显。以上结果表明抗结核治疗过程中外周淋巴细胞抗原特异性释放IFN-γ的细胞数量与CD4^+IFN-γ^+T淋巴细胞的比例随着患者的病情好转呈现下降趋势,提示抗原特异性细胞免疫应答的动态变化谱与病原菌负荷下降存在密切关系,可能成为抗结核治疗过程中的免疫学标志物。
The current anti-tuberculosis（TB） chemotherapy includes a two months intensive period and a four months consolidate period. In the process of anti-tuberculosis treatment, the dynamic change of mycobacterial specific cellular immune response is supposed to be affected by the decrease in bacteria loads, which might become biomarkers for treatment and prognosis. In this study, 9 active TB patients have been tracked to monitor the mycobacterial antigen specific T cell responses during the treatment. Peripheral blood samples were collected before the treatment and at 2, 4 and 6 months of the treatment. Antigen specific IFN-γ secreting cells were determined by enzyme-linked immune assay spots（enzyme-linked immune spot assay, ELIS- POT）. Meanwhile, antigen specific cytokine-producing Thl cells were detected by intracellular cytokine staining and mono- and multifunctional T cells proportion were analyzed. Our results showed that along with treatment, the numbers of IFN-y secreting ceils significantly decreased from （213.1 ± 40.40 ）/2.5 ×10^5 to （ 113.5 ± 23.82 ）/2.5 ×10^5 （ P = 0.0144） upon ESAT-6 stimulation and from （106.7±22.54）/2.5× 10^5 to （34. 94±17.6）/2.5×10^5 （P=0. 0234） upon CFP-10 stimulation. This was consistent with the results from flow cytometry that the percentage of IFN-γ^＋ CD4^＋ T cells was decreased after 6- month treatment. However, TNF-α^＋ CD4^＋ T and IL-2^＋ CD4^＋T sustained at similar levels after treatment when compared to non-treat- ment points, which suggested different roles of cytokines during anti-TB immune responses. Although multifunctional T cells are reported to play an important role in anti-TB response, there exhibited no obvious change during the process of treatment in our study. Taken together, our study demonstrated that antigen specific IFN-γ release displayed a decrease tendency, which was largely due to the reduction of bacilli load after the treatment. It might become the immunological biomarkers in the process of anti-tuberc
出处
《现代免疫学》
CSCD
北大核心
2017年第3期198-205,共8页
Current Immunology
基金
"艾滋病和病毒性肝炎等重大传染病防治专项"(2013ZX10003007-003-003)
国家自然科学青年基金(81501361)
上海市免疫学研究所课题组长启动经费
