Falchetti R, Di Francesco P, Lanzilli G, Gaziano R, Casalinuovo I A, Palamara A T, Ravagnan G, Garaci E
Institute of Experimental Medicine, CNR, Rome, Italy.
Immunology. 1998 Nov;95(3):346-51. doi: 10.1046/j.1365-2567.1998.00608.x.
We have studied the patterns of interleukin-2 (IL-2), IL-4 and interferon-gamma (IFN-gamma) co-expression displayed by individual splenic CD4+ and CD8+ T cells in response to influenza virus immunization. Unseparated spleen cells obtained from mice intraperitoneally (i.p.) injected with A/PR8 (H1N1) influenza virus (PR8) were cultured for 24 hr in the presence of ultraviolet-inactivated PR8. As controls, cultures of both naive spleen cells stimulated with PR8 or of immune cells lacking the inactivated virus were used. The frequencies of CD4+ and CD8+ T cells expressing IL-2, IL-4 and IFN-gamma were determined by three-colour flow cytometric analysis of fixed and saponin-permeabilized cells fluorescent-stained for either CD4 or CD8 surface molecules and for one of the following combinations of two intracellular cytokines: IL-2/IL-4, IL-2/IFN-gamma and IL-4/IFN-gamma. The results showed that immunization with influenza virus induces in both CD4+ and CD8+ T cells a heterogeneity of cytokine response patterns that do not follow the type 1/type 2 polarized response model, but with substantial differences between the two populations. In fact, the analysis of the phenotypes of virus-immune CD8+ T cells revealed similar significant proportions of cells either expressing any one of the three cytokines or co-expressing combinations of them (i.e. IL-4/IL-2, IL-4/IFN-gamma and IL-2/IFN-gamma), whereas immune CD4+ T cells were seen to express almost exclusively a single cytokine per cell. The observed patterns of cytokine production suggest that influenza virus immunization induces the expression of a type 0 cytokine pattern at both population and single cell levels in CD8+ T cells and exclusively at the population level in CD4+ T cells.
我们研究了脾脏中单个CD4⁺和CD8⁺T细胞在流感病毒免疫反应中所呈现的白细胞介素-2（IL-2）、IL-4和干扰素-γ（IFN-γ）共表达模式。从经腹腔（i.p.）注射A/PR8（H1N1）流感病毒（PR8）的小鼠中获取未分离的脾细胞，在存在紫外线灭活PR8的情况下培养24小时。作为对照，使用了用PR8刺激的未免疫脾细胞培养物或缺乏灭活病毒的免疫细胞培养物。通过对固定且经皂素通透处理的细胞进行三色流式细胞术分析来确定表达IL-2、IL-4和IFN-γ的CD4⁺和CD8⁺T细胞的频率，这些细胞用CD4或CD8表面分子以及以下两种细胞内细胞因子组合之一进行荧光染色：IL-2/IL-4、IL-2/IFN-γ和IL-4/IFN-γ。结果表明，流感病毒免疫在CD4⁺和CD8⁺T细胞中均诱导了细胞因子反应模式的异质性，这些模式并不遵循1型/2型极化反应模型，且两个群体之间存在显著差异。事实上，对病毒免疫的CD8⁺T细胞表型分析显示，表达三种细胞因子中任何一种或共表达它们的组合（即IL-4/IL-2、IL-4/IFN-γ和IL-2/IFN-γ）的细胞比例相似，而免疫CD4⁺T细胞几乎每个细胞仅表达一种细胞因子。观察到的细胞因子产生模式表明，流感病毒免疫在CD8⁺T细胞的群体和单细胞水平均诱导了0型细胞因子模式的表达，而在CD4⁺T细胞中仅在群体水平诱导了该模式的表达。
