Romani L, Mencacci A, Tonnetti L, Spaccapelo R, Cenci E, Wolf S, Puccetti P, Bistoni F
Department of Experimental Medicine, University of Perugia, Italy.
Eur J Immunol. 1994 Apr;24(4):909-15. doi: 10.1002/eji.1830240419.
By means of polymerase chain reaction-assisted mRNA amplification, we have monitored message levels of interleukin (IL)-12 in splenic macrophages and of interferon-gamma (IFN-gamma), IL-4, and IL-10 in CD4+ and CD8+ T cells using Candida albicans/host combinations that result either in a T helper type-1 (Th1)-associated self-limiting infection ("healer mice") or in a Th2-associated progressive disease ("nonhealer mice"). The timing and pattern of message detection did not differ qualitatively by the expression of IFN-gamma or IL-10 mRNA in CD4+ and CD8+ cells from healer (i.e. PCA-2 into CD2F1) vs. nonhealer (i.e. CA-6 into CD2F1 or PCA-2 into DBA/2) mice. In contrast, IL-4 mRNA was uniquely expressed by CD4+ cells from nonhealer animals. IL-12p40 was readily detected in macrophages from healer mice but was detected only early in infection in mice with progressive disease. Cytokine levels were measured in sera, and antigen-driven cytokine production by CD4+ and CD8+ cells was assessed in vitro, while IFN-gamma-producing cells were enumerated in CD4- CD8- cell fractions. Overall, our results showed that (i) antigen-specific secretion of IFN-gamma protein in vitro by CD4+ cells occurred only in healing infection; (ii) IL-4- and IL-10-producing CD4+ cells would expand in nonhealer mice in the face of high levels of circulating IFN-gamma, likely released by CD4- CD8- lymphocytes; (iii) a finely regulated IFN-gamma production correlated in the healer mice with IL-12 mRNA detection, and IL-12 was required in vitro for yeast-induced development of IFN-gamma-producing CD4+ cells. Although the mutually exclusive production of IL-4/IL-10 and IFN-gamma by early CD4+ cells may be the major discriminative factor of cure and noncure responses in candidiasis, IL-12 rather than IFN-gamma production may be an indicator of Th1 differentiation.
通过聚合酶链反应辅助的mRNA扩增技术，我们使用白色念珠菌/宿主组合监测了脾巨噬细胞中白细胞介素（IL）-12以及CD4⁺和CD8⁺ T细胞中干扰素-γ（IFN-γ）、IL-4和IL-10的信使水平，这些组合分别导致1型辅助性T细胞（Th1）相关的自限性感染（“治愈小鼠”）或Th2相关的进行性疾病（“非治愈小鼠”）。来自治愈小鼠（即PCA-2接种到CD2F1小鼠）与非治愈小鼠（即CA-6接种到CD2F1小鼠或PCA-2接种到DBA/2小鼠）的CD4⁺和CD8⁺细胞中IFN-γ或IL-10 mRNA的表达在信使检测的时间和模式上没有定性差异。相反，非治愈动物的CD4⁺细胞独特地表达IL-4 mRNA。在治愈小鼠的巨噬细胞中很容易检测到IL-12p40，但在患有进行性疾病的小鼠中仅在感染早期检测到。检测了血清中的细胞因子水平，并在体外评估了CD4⁺和CD8⁺细胞由抗原驱动的细胞因子产生，同时在CD4⁻CD8⁻细胞组分中计数产生IFN-γ的细胞。总体而言，我们的结果表明：（i）CD4⁺细胞在体外抗原特异性分泌IFN-γ蛋白仅发生在治愈性感染中；（ii）在面对可能由CD4⁻CD8⁻淋巴细胞释放的高水平循环IFN-γ时，产生IL-4和IL-10的CD4⁺细胞在非治愈小鼠中会扩增；（iii）在治愈小鼠中，精细调节的IFN-γ产生与IL-12 mRNA检测相关，并且在体外酵母诱导产生IFN-γ的CD4⁺细胞的发育需要IL-12。尽管早期CD4⁺细胞相互排斥地产生IL-4/IL-10和IFN-γ可能是念珠菌病治愈和非治愈反应的主要判别因素，但IL-12的产生而非IFN-γ的产生可能是Th1分化的一个指标。
