Matsui M, Araya S, Wang H-Y, Onai N, Matsushima K, Saida T
Department of Neurology and Clinical Research Center, Center for Neurological Diseases, Utano National Hospital, Kyoto, Japan.
Clin Exp Immunol. 2003 Nov;134(2):225-31. doi: 10.1046/j.1365-2249.2003.02291.x.
To determine whether there is an association between intracellular cytokine profiles and the expression of surface antigens, we performed a simultaneous flow cytometric analysis of these laboratory parameters in 11 healthy volunteers. Peripheral blood lymphocytes were double-stained for CD4 or CD8, as well as CD11a, CD25, CD26, CD29 and CD45RA or the chemokine receptors CCR3, CCR4, CCR5 or CXCR3. Portions of the cell samples were cultured for 4 h in the presence of 1 microm monensin and 20 microg/ml brefeldin A with or without stimulation by phorbol myristate acetate plus ionomycin for the detection of intracellular interferon-gamma (IFN-gamma), interleukin-2 (IL-2), tumour necrosis factor (TNF)-alpha, and IL-4. As a result, CD4+CD29high helper inducer T cells were closely associated with IFN-gamma and TNF-alpha producing CD4+ cells, while CD4+CXCR3+ cells showed a negative correlation with IL-4-producing cells, suggesting that both of these CD4+ subsets consist mainly of Th1 cells. In contrast, CD4+CD45RA+ cells were correlated inversely with IFN-gamma and TNF-alpha-producing cells, and CD8+CD11ahigh killer effector and total CCR5+ cells showed an inverse correlation with IL-2 producing cells, suggesting an immunoregulatory role for these three subsets in non-pathological conditions. Therefore, monitoring of lymphocyte subsets that express functional surface antigens could provide additional information concerning immune deviation, as assessed by the production of Th1/Th2 type cytokines. Further, this type of combined study may provide clues for the pathogenesis of immune-mediated disorders.
为了确定细胞内细胞因子谱与表面抗原表达之间是否存在关联，我们对11名健康志愿者的这些实验室参数进行了同步流式细胞术分析。外周血淋巴细胞用CD4或CD8以及CD11a、CD25、CD26、CD29和CD45RA或趋化因子受体CCR3、CCR4、CCR5或CXCR3进行双染色。将部分细胞样本在存在1微摩尔莫能菌素和20微克/毫升布雷菲德菌素A的情况下培养4小时，有或没有佛波醇肉豆蔻酸酯乙酸盐加离子霉素的刺激，以检测细胞内干扰素-γ（IFN-γ）、白细胞介素-2（IL-2）、肿瘤坏死因子（TNF）-α和IL-4。结果，CD4+CD29高辅助诱导T细胞与产生IFN-γ和TNF-α的CD4+细胞密切相关，而CD4+CXCR3+细胞与产生IL-4的细胞呈负相关，这表明这两个CD4+亚群主要由Th1细胞组成。相反，CD4+CD45RA+细胞与产生IFN-γ和TNF-α的细胞呈负相关，CD8+CD11a高杀伤效应细胞和总CCR5+细胞与产生IL-2的细胞呈负相关，这表明这三个亚群在非病理条件下起免疫调节作用。因此，监测表达功能性表面抗原的淋巴细胞亚群可以提供有关免疫偏差的额外信息，这是通过Th1/Th2型细胞因子的产生来评估的。此外，这种类型的联合研究可能为免疫介导疾病的发病机制提供线索。
