Impaired expression of perforin and granulysin in CD8+ T cells at the site of infection in human chronic pulmonary tuberculosis
Impaired expression of perforin and granulysin in CD8+ T cells at the site of infection in human chronic pulmonary tuberculosis
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DOI：
10.1128/iai.00624-07
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发表时间：
2007-11-01
影响因子：
3.1
通讯作者：
Grundstroem, Susanna
中科院分区：
文献类型：
--
作者：
Andersson, Jan;Samarina, Arina;Grundstroem, Susanna
结核病中的保护性免疫依赖于从效应T细胞协调释放溶细胞效应分子和随后的颗粒相关的对感染靶细胞的杀伤。在这项研究中，我们研究了慢性进行性结核病患者冷冻保存的肺组织中单细胞水平的溶细胞分子（穿孔素和颗粒酶A）和抗菌分子（颗粒溶素）的表达。通过原位成像进行蛋白表达细胞的定量，而通过实时PCR分析感染组织中的mRNA水平。在所有患者的结核病灶中，持续性炎症，包括CD 68（+）巨噬细胞中诱导型一氧化氮合酶的过度表达和CD 3（+）、CD 8（+）和CD 4（+）T细胞的显著浸润，均明显。然而，尽管CD 3（+）T细胞的积累，穿孔素和颗粒溶素表达的CD 3（+）T细胞在结核病病灶中的比例分别比远端肺实质和未感染的对照肺低2 - 3倍。这在蛋白质和mRNA水平上都很明显。此外，穿孔素和颗粒溶素表达的CD 8（+）T细胞在结核病灶内的单个肉芽肿中很少。相反，在所有患者的病变中，表达颗粒酶A的CD 3（+）T细胞明显上调。共聚焦显微镜显示穿孔素和颗粒溶素共表达，主要在CD 8（+）T细胞中;然而，这种表达在结核病病变中较低。这些发现表明，有症状的慢性结核病与感染局部CD 8（+）T细胞中穿孔素和颗粒溶解素共表达上调不足有关。
Protective immunity in tuberculosis is dependent on the coordinated release of cytolytic effector molecules from effector T cells and the subsequent granule-associated killing of infected target cells. In this study, we investigated the expression of cytolytic (perforin and granzyme A) and antimicrobial (granulysin) molecules at the single-cell level in cryopreserved lung tissue from patients with chronic, progressive tuberculosis disease. Quantification of protein-expres sing cells was performed by in situ imaging, while mRNA levels in the infected tissue were analyzed by real-time PCR. Persistent inflammation, including excessive expression of inducible nitric oxide synthase in CD68(+) macrophages and significant infiltration of CD3(+), CD8(+) and CD4(+) T cells, was evident in tuberculosis lesions in all patients. However, despite the accumulation of CD3(+) T cells, perforin-and granulysin-expressing CD3(+) T cells were detected at two- to threefold-lower ratios in the tuberculosis lesions than in distal lung parenchyma and uninfected control lungs, respectively. This was evident at both the protein and mRNA levels. Moreover, perforin- and granulysin-expressing CD8(+) T cells were scarce in individual granulomas within the tuberculosis lesions. In contrast, significant up-regulation of granzyme A-expressing CD3(+) T cells was evident in the lesions from all patients. Confocal microscopy revealed coexpression of perforin and granulysin, primarily in CD8(+) T cells; however, this expression was lower in the tuberculosis lesions. These findings suggest that symptomatic, chronic tuberculosis disease is associated with insufficient up-regulation of perforin and granulysin coexpression in CD8(+) T cells at the local site of infection.