Bjorndahl J M, Sung S S, Hansen J A, Fu S M
Immunology Program, Oklahoma Medical Research Foundation, Oklahoma City.
Eur J Immunol. 1989 May;19(5):881-7. doi: 10.1002/eji.1830190515.
Monoclonal antibodies (mAb) against CD3 or CD28 in conjunction with the tumor promoter phorbol 12-myristate 13-acetate (PMA) induce interleukin 2 receptor (IL2R) expression, IL2 production and proliferation in resting T cells. Recent studies indicate that these two pathways are biochemically distinct. In this study T cell activation induced by PMA and anti-CD28 mAb 9.3 is compared to the effects of PMA plus anti-CD3 mAb (T3-II and 235) in the presence or absence of cyclosporin A (CsA), dibutyryladenosine 3':5' cyclic monophosphate (db-cAMP) or cholera toxin (CT). Proliferation of T cells stimulated with PMA plus mAb 9.3 is resistant to the inhibitory effects of CsA, db-cAMP and CT. Only at the highest dose did CsA have any effect on PMA plus mAb 9.3-induced T cell proliferation. Conversely, CsA, db-cAMP and CT inhibit PMA plus T3-II-induced T cell proliferation. mRNA analysis further demonstrates the similarities and the differences between the CD28 and CD3 activation pathways. Recently, T3-II was reported to induce tumor necrosis factor (TNF) and lymphotoxin (LT) mRNA synthesis in PMA-treated T cells. In this study mAb 9.3 is shown to substitute for T3-II in the induction of TNF and mRNA. However, the production of TNF and LT mRNA in PMA plus mAb 9.3-treated T cells is greater than that seen in PMA plus T3-II-treated cells. mRNA synthesis included by PMA plus T3-II is blocked by CsA. mRNA production in T cells activated with PMA plus mAb 9.3 is resistant to CsA. Similar results are noted with IL2 and IL2R mRNA. Flow cytometric analysis of the IL2R confirms the mRNA data. CsA blocks the T3-II-induced potentiation of PMA-induced IL2R expression but not the mAb 9.3-induced potentiation. This differential inhibitory effect of CsA on IL2R expression is also seen with db-cAMP and CT. We examined the effects of these two pathways on the expression of the early activation antigen EA 1 and cytoplasmic free calcium. Recently, we have shown anti-CD3 mAb potentiate EA 1 expression induced by 1,2-sn-dioctanoylglycerol and this potentiation is calcium dependent. dp-cAMP blocks T3-II- and 235-induced potentiation of EA1 expression and inhibits the T3-II- and 235-mediated rise in intracellular free calcium [( Ca2+]i). Conversely, 9.3 does not potentiate EA 1 expression or induce a rise in [Ca2+]i. These results provide further evidence that the CD28 and CD3 activation pathways utilize distinct signal transduction pathways.
抗CD3或CD28的单克隆抗体（mAb）与肿瘤启动子佛波醇12 - 肉豆蔻酸酯13 - 乙酸酯（PMA）联合使用可诱导静息T细胞中白细胞介素2受体（IL2R）表达、IL2产生及细胞增殖。最近的研究表明这两条途径在生化方面是不同的。在本研究中，将PMA和抗CD28 mAb 9.3诱导的T细胞活化与PMA加抗CD3 mAb（T3 - II和235）在存在或不存在环孢素A（CsA）、二丁酰腺苷3':5'环一磷酸（db - cAMP）或霍乱毒素（CT）的情况下的作用进行比较。用PMA加mAb 9.3刺激诱导的T细胞增殖对CsA、db - cAMP和CT的抑制作用具有抗性。只有在最高剂量时，CsA才对PMA加mAb 9.3诱导的T细胞增殖有任何作用。相反，CsA、db - cAMP和CT抑制PMA加T3 - II诱导的T细胞增殖。mRNA分析进一步证明了CD28和CD3激活途径之间的异同。最近，有报道称T3 - II可诱导PMA处理的T细胞中肿瘤坏死因子（TNF）和淋巴毒素（LT）mRNA的合成。在本研究中，mAb 9.3在诱导TNF和mRNA方面可替代T3 - II。然而，PMA加mAb 9.3处理的T细胞中TNF和LT mRNA的产生量大于PMA加T3 - II处理的细胞。PMA加T3 - II诱导的mRNA合成被CsA阻断。用PMA加mAb 9.3激活的T细胞中的mRNA产生对CsA具有抗性。IL2和IL2R mRNA也有类似结果。对IL2R的流式细胞术分析证实了mRNA数据。CsA阻断T3 - II诱导的PMA诱导的IL2R表达增强，但不阻断mAb 9.3诱导的增强作用。db - cAMP和CT对IL2R表达也有这种不同的抑制作用。我们研究了这两条途径对早期激活抗原EA 1表达和细胞质游离钙的影响。最近，我们已表明抗CD3 mAb增强由1,2 - 二辛酰甘油诱导的EA 1表达，且这种增强是钙依赖性的。dp - cAMP阻断T
