Johnson Barbara J, Costelloe Elaine O, Fitzpatrick David R, Haanen John B A G, Schumacher Ton N M, Brown Lorena E, Kelso Anne
Cooperative Research Centre for Vaccine Technology and Queensland Institute of Medical Research, Post Office Royal Brisbane Hospital, Queensland 4029, Australia.
Proc Natl Acad Sci U S A. 2003 Mar 4;100(5):2657-62. doi: 10.1073/pnas.0538056100. Epub 2003 Feb 24.
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 B, 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.
流感病毒感染会激活细胞毒性T淋巴细胞（CTL），这些细胞通过从细胞质颗粒中释放穿孔素和颗粒酶来促进病毒清除。在初次流感病毒感染期间，从新鲜分离的小鼠肺实质细胞中可检测到病毒特异性、依赖穿孔素的CD8(+) CTL，但在其启动部位纵隔淋巴结（MLN）或脾脏中未检测到。为了确定这种差异是由于MLN中效应CTL的频率低还是成熟不完全所致，我们在从病毒感染小鼠分离后立即测量了CD8(+)群体和单细胞中穿孔素、颗粒酶A、B和C以及IFN-γ mRNA的表达。定量PCR显示，来自MLN、脾脏和肺实质的活化CD8(+)细胞中穿孔素、颗粒酶A、颗粒酶B和IFN-γ有显著表达。未检测到颗粒酶C的表达。来自这三个组织的单个活化或与核蛋白肽/ I类四聚体结合的CD8(+)细胞表达了穿孔素、颗粒酶和IFN-γ mRNA的不同组合。虽然肺中的细胞表达颗粒酶A和B的频率较高，但每个组织中都含有同时表达穿孔素与颗粒酶A和/或B的细胞。MLN和肺之间的主要差异在于肺中活化CD8(+) T细胞的频率升高，而不是它们的穿孔素/颗粒酶表达谱。数据表明，一些CTL在MLN中成熟为表达穿孔素/颗粒酶的效应细胞，但只有当它们在感染的肺中积累时才能达到可检测的频率。
