Andersson U, Andersson J, Lindfors A, Wagner K, Möller G, Heusser C H
Department of Immunology, University of Stockholm, Sweden.
Eur J Immunol. 1990 Jul;20(7):1591-6. doi: 10.1002/eji.1830200727.
The production of interleukin 2 (IL 2), IL 4 and interferon-gamma (IFN-gamma) by in vitro activated unselected human blood mononuclear cells was studied at a single-cell level. Individual lymphokine-synthesizing cells were identified by intracellular immunofluorescent staining using cytokine-specific monoclonal or polyclonal antibodies. Cultures from adult blood donors revealed a biphasic kinetic production pattern for IL 2 and IFN-gamma with peaks occurring 4-6 and 24-30 h after initiation of the cultures. Approximately 20%-40% of the lymphocytes produced IL 2 and IFN-gamma. In contrast, only 1%-3% of the lymphocytes synthesized IL 4 with maximal frequency after 6 h of culture. CD4+ as well as CD8+ T cells contributed to the synthesis of all three lymphokines studied. CD4+CD45R- T cells were the major producers of IL 2 and IL 4, while CD8+CD45R- T cells were the most common phenotype of IFN-gamma-synthesizing cells. By performing two-color immunofluorescence studies we observed that among IL 4-producing cells every second one made simultaneously IL 2 and every fourth one made IFN-gamma. Mononuclear cells from umbilical cord blood could be stimulated to make IL 2 to the same extent as cells from adult blood donors. No IL 4 production and a strikingly reduced frequency of IFN-gamma producers were noted in cell cultures from neonates. IL 2, IL 4 and IFN-gamma accumulated in the Golgi system, which resulted in a characteristic morphology of the staining, eliminating problems with evaluation of background signals.
在单细胞水平上研究了体外激活的未分选人血单核细胞产生白细胞介素2（IL-2）、IL-4和干扰素-γ（IFN-γ）的情况。使用细胞因子特异性单克隆或多克隆抗体通过细胞内免疫荧光染色鉴定单个淋巴因子合成细胞。来自成年献血者的培养物显示IL-2和IFN-γ的双相动力学产生模式，在培养开始后4 - 6小时和24 - 30小时出现峰值。大约20% - 40%的淋巴细胞产生IL-2和IFN-γ。相比之下，只有1% - 3%的淋巴细胞在培养6小时后以最高频率合成IL-4。CD4⁺以及CD8⁺T细胞都参与了所研究的三种淋巴因子的合成。CD4⁺CD45R⁻T细胞是IL-2和IL-4的主要产生者，而CD8⁺CD45R⁻T细胞是合成IFN-γ细胞中最常见的表型。通过进行双色免疫荧光研究，我们观察到在产生IL-4的细胞中，每第二个细胞同时产生IL-2，每第四个细胞产生IFN-γ。脐血单核细胞可以被刺激产生与成年献血者细胞相同程度的IL-2。在新生儿的细胞培养物中未观察到IL-4的产生，并且产生IFN-γ的细胞频率显著降低。IL-2、IL-4和IFN-γ在高尔基体系统中积累，这导致了染色的特征性形态，消除了背景信号评估的问题。
