Single-cell perforin and granzyme expression reveals the anatomical localization of effector CD8+ T cells in influenza virus-infected mice
Single-cell perforin and granzyme expression reveals the anatomical localization of effector CD8+ T cells in influenza virus-infected mice
复制标题
DOI：
10.1073/pnas.0538056100
复制
发表时间：
2003-03-04
影响因子：
11.1
通讯作者：
Kelso, A
中科院分区：
文献类型：
--
作者：
Johnson, BJ;Costelloe, EO;Kelso, A
流感病毒感染激活溶细胞性T淋巴细胞（CTL），其通过从细胞质颗粒释放穿孔素和颗粒酶而有助于病毒清除。在小鼠肺实质新鲜分离的细胞中检测到病毒特异性穿孔素依赖性CD 8（+）CTL，但在纵隔淋巴结（MLN）中未检测到病毒特异性穿孔素依赖性CD 8（+）CTL，在纵隔淋巴结（MLN）中未检测到病毒特异性穿孔素依赖性CD 8（+）CTL，或在原发性流感病毒感染期间未检测到病毒特异性穿孔素依赖性CD 8（+）CTL。为了确定这种差异是否是由于MLN中效应CTL的低频率或不完全成熟，我们在从病毒感染小鼠分离后立即测量了CD 8（+）群体和单个细胞中穿孔素、颗粒酶A、B和C以及IFN-γ mRNA的表达。定量PCR显示MLN、脾和肺实质中活化的CD 8（+）细胞中穿孔素、颗粒酶A、颗粒酶13和IFN-γ的显著表达。未检测到颗粒酶C表达。来自三种组织的单个活化或核蛋白肽/I类四聚体结合的CD 8（+）细胞表达穿孔素、颗粒酶和IFN-γ mRNA的不同组合。尽管来自肺的细胞以较高频率表达颗粒酶A和B，但每个组织都含有与颗粒酶A和/或B共表达穿孔素的细胞。MLN和肺之间的主要差异是肺中活化的CD 8（+）T细胞的频率升高，而不是它们的穿孔素/颗粒酶表达谱。这些数据表明，一些CTL成熟成穿孔素/颗粒酶表达的效应细胞在MLN，但达到可检测的频率，只有当他们在感染的肺积累。
Influenza virus infection activates cytolytic T lymphocytes (CTL) that contribute to viral clearance by releasing perforin and granzymes from cytoplasmic granules. Virus-specific, perforin-dependent CD8(+) CTL were detected in freshly isolated cells from the mouse lung parenchyma but not from the mediastinal lymph nodes (MLN), where they are primed, or from the spleen during primary influenza virus infection. To determine whether this difference was due to the low frequency or incomplete maturation of effector CTL in MLN, we measured expression of perforin, granzymes A, B, and C, and IFN-gamma mRNAs in CD8(+) populations and single cells immediately after isolation from virus-infected mice. Quantitative PCR revealed significant expression of perforin, granzyme A, granzyme 13, and IFN-gamma in activated CD8(+) cells from MLN, spleen, and lung parenchyma. Granzyme C expression was not detected. Individual activated or nucleoprotein peptide/class I tetramer-binding CD8(+) cells from the three tissues expressed diverse combinations of perforin, granzyme, and IFN-gamma mRNAs. Although cells from lung expressed granzymes A and B at higher frequency, each of the tissues contained cells that coexpressed perforin with granzymes A and/or B. The main difference between MLN and lung was the elevated frequency of activated CD8(+) T cells in the lung, rather than their perforin/granzyme expression profile. The data suggest that some CTL mature into perforin/granzyme-expressing effector cells in MLN but reach detectable frequencies only when they accumulate in the infected lung.