Degiannis D, Luke-Gustites D, Mikhail N, Raska K, Raskova J
University Diagnostic Laboratories, University of Medicine and Dentistry, New Jersey-Robert Wood Johnson Medical School, Piscataway 08854.
Transplantation. 1992 Aug;54(2):308-12. doi: 10.1097/00007890-199208000-00021.
We studied the effect of cyclosporine A, prednisolone, and the Ca2+ channel blocker verapamil on interleukin-6 binding to mitogen-activated peripheral blood mononuclear cells, using a flow cytometric technique and phycoerythrin-conjugated IL-6. All mitogenic stimuli up-regulated IL-6 binding to a variable degree. PHA alone or in combination with PMA was the most effective stimulant in up-regulating IL-6 binding in all the experiments performed. The main changes in IL-6 binding were seen in the large cell cluster, which consisted mainly of lymphoblasts. PHA and PHA/PMA, however, also up-regulated the mean fluorescence intensity on the small cell cluster, which consisted mainly of quiescent lymphocytes. The overall effect of the three pharmacological agents on mitogen-up-regulated IL-6 binding was minimal; most significant were a down-regulation by all three agents of IL-6 binding by small lymphocytes in PHA/PMA cultures, a down-regulation of IL-6 binding by CsA in PHA/PMA-induced large PBMC, and an up-regulation by verapamil of PMA-induced IL-6 binding in large PBMC. Measurements of IL-2 binding and of IL-6 production in the same cultures showed a different pattern than that seen with IL-6 binding, as well as different CsA, prednisolone, and verapamil action. In conclusion, by using a new flow cytometric technique providing information both about the quantity of bound cytokine and about the proportion of IL-6-binding cells, we have demonstrated that IL-6 receptor expression in vitro by PBMC can be up-regulated by the use of stimulants differing in the signal transduction pathways they activate. In addition, by using different pharmacological agents and stimuli to dissect different activation pathways of the in vitro immune response, we conclude that IL-6R generation is regulated differently from IL-6 production. Furthermore, since CsA and prednisolone are known inhibitors of in vitro IL-2 production, our results indicate that IL-6R generation does not rely exclusively on the presence of IL-2.
我们使用流式细胞术和藻红蛋白偶联的白细胞介素-6（IL-6），研究了环孢素A、泼尼松龙和钙离子通道阻滞剂维拉帕米对IL-6与丝裂原激活的外周血单个核细胞结合的影响。所有促有丝分裂刺激均不同程度地上调了IL-6的结合。在所有进行的实验中，单独使用PHA或与PMA联合使用是上调IL-6结合最有效的刺激物。IL-6结合的主要变化见于主要由淋巴母细胞组成的大细胞群。然而，PHA和PHA/PMA也上调了主要由静止淋巴细胞组成的小细胞群的平均荧光强度。这三种药物对丝裂原上调的IL-6结合的总体影响很小；最显著的是，在PHA/PMA培养物中，这三种药物均下调了小淋巴细胞的IL-6结合，环孢素A在PHA/PMA诱导的大的外周血单个核细胞（PBMC）中下调了IL-6结合，而维拉帕米在大的PBMC中上调了PMA诱导的IL-6结合。在相同培养物中对IL-2结合和IL-6产生的测量显示出与IL-6结合不同的模式，以及环孢素A、泼尼松龙和维拉帕米的不同作用。总之，通过使用一种新的流式细胞术技术，该技术既能提供关于结合的细胞因子数量的信息，又能提供关于IL-6结合细胞比例的信息，我们证明了PBMC在体外的IL-6受体表达可通过使用激活不同信号转导途径的刺激物而上调。此外，通过使用不同的药物和刺激物来剖析体外免疫反应的不同激活途径，我们得出结论，IL-6受体的产生与IL-6的产生受到不同的调节。此外，由于环孢素A和泼尼松龙是已知的体外IL-2产生的抑制剂，我们的结果表明IL-6受体的产生并不完全依赖于IL-2的存在。
