Harada Y, Watanabe S, Yssel H, Arai K
Department of Molecular and Developmental Biology, University of Tokyo, Japan.
J Allergy Clin Immunol. 1996 Dec;98(6 Pt 2):S161-73. doi: 10.1016/s0091-6749(96)70063-5.
According to the widely accepted classification, human TH cell clones can be divided into two mutually exclusive subsets, TH1 and TH2, based on their profile of cytokine production. The intracellular difference between these clones is not clear. To characterize the biochemical nature of T-cell receptor (TCR)/CD3 complex-mediated signal transduction pathways, we introduced several human TH cell clones of THO- or TH1-like phenotype and analyzed the effects of various drugs and antibodies on cytokine production or proliferation of these clones. The tyrosine kinase inhibitor herbimycin inhibited the production of interferon-gamma (IFN-gamma) by THO-like clone, after stimulation with anti-CD3 monoclonal antibody alpha CD3-mAb) or with phorbol 12-myristate 13-acetate (PMA) and the calcium ionophore A23187. However, whereas herbimycin strongly inhibited the production of IL-4 and IL-5 by alpha CD3 mAb stimulated T cells, it did not affect the production of these cytokines after PMA/A23187 stimulation. Cyclosporin A inhibited the proliferation as well as the production of the cytokines, including that of IL-2, IL-4, IL-5, and IFN-gamma, irrespective of the mode of stimulation. A23187, which synergizes with PMA in the induction of IL-4 and IFN-gamma, inhibited PMA-induced IL-10 production in a dose-dependent manner. Transforming growth factor-beta and anti-IL-2 receptor monoclonal antibody partially inhibited alpha CD3 mAb-mediated T-cell proliferation, but had no effect on the proliferation induced by PMA and A23187. Cyclic adenosine monophosphate (cAMP)-elevating drugs, like prostaglandin E2 and dibutyryl cAMP, inhibited the TCR-mediated cytokine production but shifted the cytokine production profile from a TH0 to a TH2 type after stimulation with PMA and A23187. Finally, we analyzed the induction of activity of two transcription factors, nuclear factor-kappa B (NF-kappa B) and nuclear factor of activated T cells, involved in the regulation of cytokine gene expression, after a different mode of activation. The induction of NF-kappa B (p50/p65 heterodimer) by using alpha CD3-mAb stimulation but not by using PMA/A23187 stimulation was found to be inhibited by using cAMP-elevating drugs.
根据广泛接受的分类，人类TH细胞克隆可根据其细胞因子产生谱分为两个相互排斥的亚群，即TH1和TH2。这些克隆之间的细胞内差异尚不清楚。为了表征T细胞受体（TCR）/CD3复合物介导的信号转导途径的生化性质，我们引入了几种TH0或TH1样表型的人类TH细胞克隆，并分析了各种药物和抗体对这些克隆的细胞因子产生或增殖的影响。酪氨酸激酶抑制剂赫伯霉素在抗CD3单克隆抗体（αCD3 - mAb）或佛波醇12 - 肉豆蔻酸酯乙酸酯（PMA）和钙离子载体A23187刺激后，抑制了TH0样克隆产生干扰素 - γ（IFN - γ）。然而，尽管赫伯霉素强烈抑制αCD mAb刺激的T细胞产生IL - 4和IL - 5，但它对PMA/A23187刺激后这些细胞因子的产生没有影响。环孢素A抑制增殖以及包括IL - 2、IL - 4、IL - 5和IFN - γ在内的细胞因子的产生，而与刺激方式无关。与PMA协同诱导IL - 4和IFN - γ的A23187以剂量依赖性方式抑制PMA诱导的IL - 10产生。转化生长因子 - β和抗IL - 2受体单克隆抗体部分抑制αCD3 mAb介导的T细胞增殖，但对PMA和A23187诱导的增殖没有影响。环磷酸腺苷（cAMP）升高药物，如前列腺素E2和二丁酰cAMP，抑制TCR介导的细胞因子产生，但在PMA和A23187刺激后将细胞因子产生谱从TH0型转变为TH2型。最后，我们分析了在不同激活模式后，参与细胞因子基因表达调控的两种转录因子，核因子 - κB（NF - κB）和活化T细胞核因子的活性诱导情况。发现使用cAMP升高药物可抑制通过αCD3 - mAb刺激而非PMA/A23187刺激诱导的NF - κB（p50/p65异二聚体）。
