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Expression of the interleukin-4 receptor alpha in human conjunctival epithelial cells.

机译:白细胞介素4受体α在人结膜上皮细胞中的表达。

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AIM: To investigate the expression and function of interleukin-4 receptor alpha (IL-4Ralpha) in human conjunctival epithelial cells (HCjECs). METHODS: The presence of IL-4Ralpha mRNA and protein was examined by reverse transcription (RT) PCR and immunohistology, respectively. Cell surface expression was examined by flow cytometry. The effects of interleukin (IL)-4 or IL-13 on the tyrosine phosphorylation of signal transducer and the activator of transcription 6 (STAT6) were evaluated by immunoblot analyses. The transcripts upregulated upon IL-4 stimulation were examined using GeneChip, and confirmed by quantitative RT-PCR. RESULTS: IL-4Ralpha mRNA and protein were detected in human conjunctival epithelium. IL-4Ralpha protein was expressed on the cell surface of HCjECs. IL-4 and IL-13 induced tyrosine phosphorylation of STAT6. GeneChip analysis showed that nine transcripts were upregulated more than fourfold by IL-4 stimulation in the primary HCjECs from two individuals. Quantitative RT-PCR assay confirmed the upregulation of these transcripts: lecithin retinol acyltransferase (LRAT), calpain (CAPN14), tumour necrosis factor alpha-induced protein 6 (TNFAIP6), RAS guanyl-releasing protein 1 (RASGRP1), endothelin receptor type A (EDNRA), hyaluronan synthase 3 (HAS3), cathepsin C (CTSC), carbonic anhydrase II (CA2) and cytokine-inducible SH2-containing protein (CISH). CONCLUSIONS: HCjECs expressed functioning IL-4Ralpha, and IL-4 stimulation induced the expression of several genes.
机译:目的:研究白细胞介素4受体α(IL-4Ralpha)在人结膜上皮细胞(HCjECs)中的表达和功能。方法:分别通过逆转录(RT)PCR和免疫组织学检查IL-4Ralpha mRNA和蛋白的存在。通过流式细胞术检查细胞表面表达。通过免疫印迹分析评估白介素(IL)-4或IL-13对信号转导子和转录激活因子6(STAT6)酪氨酸磷酸化的影响。使用GeneChip检查IL-4刺激后上调的转录本,并通过定量RT-PCR进行确认。结果:在人结膜上皮中检测到IL-4Ralpha mRNA和蛋白。 IL-4Ralpha蛋白在HCjECs的细胞表面表达。 IL-4和IL-13诱导STAT6的酪氨酸磷酸化。基因芯片分析显示,在来自两个个体的主要HCjEC中,IL-4刺激使9个转录本上调了四倍以上。定量RT-PCR分析证实了这些转录本的上调:卵磷脂视黄醇酰基转移酶(LRAT),钙蛋白酶(CAPN14),肿瘤坏死因子α诱导蛋白6(TNFAIP6),RAS鸟苷释放蛋白1(RASGRP1),内皮素A型受体(EDNRA),透明质酸合酶3(HAS3),组织蛋白酶C(CTSC),碳酸酐酶II(CA2)和细胞因子诱导的含SH2的蛋白质(CISH)。结论:HCjECs表达功能性IL-4Ralpha,IL-4刺激诱导了几个基因的表达。

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