Basok Anna, Shnaider Alla, Man Limor, Chaimovitz Cidio, Douvdevani Amos
Department of Nephrology, Soroka University Medical Center, Ben-Gurion University of the Negev, Faculty of Health Sciences, Beer-Sheva, Israel.
J Am Soc Nephrol. 2001 Apr;12(4):695-702. doi: 10.1681/ASN.V124695.
Limited data are available concerning the interaction between lymphocytes and human peritoneal mesothelial cells (HPMC) during peritonitis. CD40 is a member of the tumor necrosis factor (TNF) family of receptors whose ligand (CD154) is mainly expressed on the membrane of activated CD4-positive lymphocytes. CD154-CD40 cross-linking is a central event in antigen presentation, B-cell activation by T cells, and regulation of cytokine secretion from various types of cells. The goal of this study was to demonstrate in vitro the presence of CD40 on HPMC and to test its functionality in inducing interleukin-15 (IL-15) and RANTES. We assayed the levels of CD40 by reverse transcription-PCR and flow cytometry and IL-15 and RANTES by enzyme-linked immunosorbent assay. Genetically modified L cells that express elevated levels of CD154 (CD40L cells) were used to stimulate CD40. HPMC express CD40 mRNA and protein. After stimulation with interferon-gamma (IFNgamma, 5U/ml) or TNFalpha (1 ng/ml), there was a small increase in CD40 mRNA and protein levels; when both cytokines were applied, the increase in CD40 levels was more than threefold. CD40 ligation induced IL-15 production by HPMC and was additive to IFNgamma stimulation. CD40 ligation was strongly synergistic with IFNgamma in induction of RANTES (20-fold as compared with unstimulated HPMC), whereas neither ligation nor IFNgamma alone could induce RANTES. Pretreatment of HPMC with TNFalpha and IFNgamma increased the response to CD40 ligation in magnitudes that correlated with the elevation of CD40 levels induced by the pretreatment. To conclude, the presence of a functional CD40 on HPMC whose ligation induced IL-15 and RANTES production was detected. It is possible that this receptor acts as a major mediator of T-cell-regulated immune and inflammatory response during peritonitis.
关于腹膜炎期间淋巴细胞与人类腹膜间皮细胞（HPMC）之间的相互作用，可用数据有限。CD40是肿瘤坏死因子（TNF）受体家族的成员，其配体（CD154）主要表达于活化的CD4阳性淋巴细胞膜上。CD154-CD40交联是抗原呈递、T细胞激活B细胞以及调节各种细胞因子分泌的核心事件。本研究的目的是在体外证明HPMC上存在CD40，并测试其在诱导白细胞介素-15（IL-15）和调节激活正常T细胞表达和分泌的趋化因子（RANTES）方面的功能。我们通过逆转录聚合酶链反应（RT-PCR）和流式细胞术检测CD40水平，通过酶联免疫吸附测定法检测IL-15和RANTES水平。使用表达高水平CD154的基因改造L细胞（CD40L细胞）刺激CD40。HPMC表达CD40 mRNA和蛋白。用γ干扰素（IFNγ，5U/ml）或肿瘤坏死因子α（TNFα，1ng/ml）刺激后，CD40 mRNA和蛋白水平略有增加；当两种细胞因子同时应用时，CD40水平的增加超过三倍。CD40连接诱导HPMC产生IL-15，并与IFNγ刺激相加。CD40连接在诱导RANTES方面与IFNγ强烈协同（与未刺激的HPMC相比增加20倍），而单独的连接或IFNγ都不能诱导RANTES。用TNFα和IFNγ预处理HPMC增加了对CD40连接的反应程度，这与预处理诱导的CD40水平升高相关。总之，检测到HPMC上存在功能性CD40，其连接可诱导IL-15和RANTES产生。在腹膜炎期间，该受体可能作为T细胞调节的免疫和炎症反应的主要介质发挥作用。
