Gibbs V C, Wood D M, Garovoy M R
Hum Immunol. 1985 Nov;14(3):259-69. doi: 10.1016/0198-8859(85)90233-2.
We have developed in culture for the first time one of the actual target cells in kidney rejection by establishing long-term cultures of human kidney capillary endothelium and have begun to explore their response to immunologic stimuli. Microvascular endothelial cells were obtained from the human kidney cortex by mechanical separation, enzyme digestion, and density gradient centrifugation. The cells were exposed to 1% PHA, a PHA-stimulated PBL supernatant, human and mouse IL-1 and human gamma-interferon. They were stained with FITC-conjugated monoclonal antibodies specific for beta 2-microglobulin, HLA-DR, and DQ and analyzed by flow cytometry (FACS-II). HKCE cells express HLA-ABC antigens during growth and confluent phases but do not express DR or DQ antigens. Only the PHA-stimulated PBL supernatant and gamma-interferon induced class II antigen expression. A change in cellular morphology was noted after 72 hr of incubation in the presence of the lectin-stimulated PBL supernatant. Our findings show that microvascular endothelial cells express selected Ia antigens only after immunologic induction. This is in agreement with studies using umbilical and aortic endothelial cells. These data suggest that a dynamic interaction takes place between activated lymphocytes whose products induce expression of class II antigens and human kidney endothelial cells.
我们首次通过建立人肾毛细血管内皮细胞的长期培养体系，在体外培养出了肾移植排斥反应中的一种实际靶细胞，并开始探索它们对免疫刺激的反应。微血管内皮细胞是通过机械分离、酶消化和密度梯度离心从人肾皮质中获取的。将这些细胞暴露于1%的PHA、PHA刺激的外周血淋巴细胞（PBL）上清液、人和小鼠白细胞介素-1（IL-1）以及人γ-干扰素中。用与β2-微球蛋白、HLA-DR和DQ特异性结合的异硫氰酸荧光素（FITC）标记的单克隆抗体对细胞进行染色，并通过流式细胞术（FACS-II）进行分析。人肾毛细血管内皮细胞（HKCE细胞）在生长和汇合阶段表达HLA-ABC抗原，但不表达DR或DQ抗原。只有PHA刺激的PBL上清液和γ-干扰素能诱导II类抗原表达。在凝集素刺激的PBL上清液存在的情况下孵育72小时后，观察到细胞形态发生了变化。我们的研究结果表明，微血管内皮细胞仅在免疫诱导后才表达特定的Ia抗原。这与使用脐静脉和主动脉内皮细胞的研究结果一致。这些数据表明，其产物诱导II类抗原表达的活化淋巴细胞与人肾内皮细胞之间发生了动态相互作用。
