Goletti Delia, Butera Ornella, Bizzoni Federica, Casetti Rita, Girardi Enrico, Poccia Fabrizio
Second Division of Infectious Diseases of the Health Department, Department of Experimental Research, National Institute for Infectious Diseases Lazzaro Spallanzani, Rome, Italy.
J Infect Dis. 2006 Oct 1;194(7):984-92. doi: 10.1086/507427. Epub 2006 Aug 30.
Interferon (IFN)-gamma response to region of difference (RD) 1 proteins (culture filtrate 10 and early secreted antigenic target 6) or overlapping peptides is a novel diagnostic marker of tuberculosis (TB) infection. Because we have recently shown that the response to certain peptides selected from RD1 allows discrimination between active TB (A-TB) and successfully treated TB (T-TB), we analyzed here the effector memory T cell profile and RD1-specific responses under the same clinical conditions.
T cell responses to RD1 antigens were analyzed in patients with either severe or mild A-TB (classified on the basis of radiological lesions) and in 2 sets of healthy control subjects--those who had been successfully treated (the T-TB control subjects) and those whose tuberculin skin test (TST) results were negative (the TST-negative control subjects). IFN-gamma -producing CD4+ effector T cells were monitored by flow-cytometric analysis and ex vivo enzyme-linked immunospot (ELISPOT) assay, whereas a "cultured" ELISPOT assay was used to determine the frequency of memory T cells.
In the patients with severe A-TB, both CD4-mediated effector memory and central memory responses to the selected RD1 peptides were almost absent, whereas these responses were found in the majority of the patients with mild A-TB. In contrast, recognition of the selected RD1 peptides was detected in the T-TB control subjects only by expanding the central memory T cell pool.
These data suggest a protective role for RD1 peptide-specific CD4+ effector T cells, which undergo clonal expansion during Mycobacterium tuberculosis replication and then a contraction phase after disease resolution, culminating in the generation of CD4+ memory T cells.
干扰素(IFN)-γ对差异区域(RD)1蛋白(培养滤液10和早期分泌性抗原靶标6)或重叠肽的反应是结核病(TB)感染的一种新型诊断标志物。由于我们最近发现,对从RD1中选择的某些肽的反应能够区分活动性结核病(A-TB)和成功治愈的结核病(T-TB),因此我们在此分析了相同临床条件下的效应记忆T细胞谱和RD1特异性反应。
在患有重度或轻度A-TB(根据放射学病变分类)的患者以及两组健康对照受试者中分析T细胞对RD1抗原的反应,这两组健康对照受试者分别是已成功治愈的(T-TB对照受试者)和结核菌素皮肤试验(TST)结果为阴性的(TST阴性对照受试者)。通过流式细胞术分析和体外酶联免疫斑点(ELISPOT)测定法监测产生IFN-γ的CD4 +效应T细胞,而“培养的”ELISPOT测定法用于确定记忆T细胞的频率。
在重度A-TB患者中,几乎不存在对所选RD1肽的CD4介导的效应记忆和中枢记忆反应,而在大多数轻度A-TB患者中发现了这些反应。相比之下,仅通过扩大中枢记忆T细胞库才能在T-TB对照受试者中检测到对所选RD1肽的识别。
这些数据表明RD1肽特异性CD4 +效应T细胞具有保护作用,其在结核分枝杆菌复制过程中经历克隆扩增,然后在疾病消退后进入收缩期,最终产生CD4 +记忆T细胞。