免疫缺陷病毒感染对恒河猴多聚免疫球蛋白受体表达的影响

发布时间：2018-04-22 14:23

  本文选题：多聚免疫球蛋白受体 + SIV/SHIV感染　； 参考：《中国疾病预防控制中心》2017年硕士论文

【摘要】：目的:多聚免疫球蛋白受体(polymeric immunoglobulin receptor,pIgR)是黏膜免疫系统的重要组成成分,不仅在IgA从固有膜到粘膜表面的转运过程中发挥关键作用,而且直接参与SIgA的构成。因此,pIgR的有效分泌是多聚免疫球蛋白(dIgA和pIgM)发挥正常粘膜防御功能的必要条件。虽然SIgA在HIV/AIDS患者中出现严重缺陷,但对pIgR表达水平与分布在免疫缺陷病毒(HIV/SIV)感染后的变化规律尚不清楚。本研究对恒河猴肠道和呼吸道粘膜中pIgR表达水平与分布在SIV/SHIV感染后的变化以及IL-17A对体外培养人支气管上皮细胞中pIgR表达的影响进行了观察,拟阐明肠道和呼吸道pIgR在免疫缺陷病毒感染后的变化规律及可能机制。为艾滋病等黏膜损伤相关疾病中粘膜免疫系统的重建提供必要的科学基础。方法:本研究利用常规和实时荧光定量RT-PCR方法检测正常和SIV/SHIV感染恒河猴粘膜组织中pIgRmRNA及其表达水平。利用Western blotting检测正常和感染恒河猴粘膜组织中pIgR蛋白及其表达水平。利用免疫荧光标记及激光扫描共聚焦显微镜观察正常和感染恒河猴粘膜组织中pIgR蛋白的分布和丰度。另外,利用体外细胞培养技术观察IL-17A对人支气管上皮细胞(HBE)中pIgR mRNA水平和pIgR蛋白水平的影响。结果:研究发现:1)正常恒河猴胃底、十二指肠、空肠、回肠、盲肠、结肠和直肠pIgR蛋白表达水平存在部位间差异,小肠和大肠的前端表达水平较高。在SIV/SHIV感染后相对应的组织中pIgR蛋白表达水平明显下降。在胃底、空肠近端、盲肠、结肠和直肠中,差异具有统计学意义,P值分别为 0.033,0.0043,0.0364,0.0252,0.0424。2)气管粘膜中pIgRmRNA和pIgR蛋白表达水平在感染后均显著下降,P值为0.0014;IL-17AmRNA表达水平在SIV/SHIV感染后也下降,并且两者(pIgR mRNA vs IL-17A mRNA)具有相关关系(正相关)。在肺组织中pIgR mRNA和IL-17AmRNA表达水平在感染后均有所上升,两者具有相关性(正相关),但pIgR蛋白表达水平在感染后下降。3)不同浓度IL-17A刺激HBE以后,pIgR mRNA的相对表达量在72 h内均呈现先增加后减少的趋势,在24h内有一个最高值出现。结论:上述结果表明,SIV/SHIV感染恒河猴消化道粘膜和气管粘膜中pIgR转录水平和蛋白表达水平都出现不同程度的降低,IL-17A对粘膜上皮细胞pIgR的表达具有一定的促进作用。免疫缺陷病毒感染后粘膜pIgR表达异常可能与IL-17A水平下降有关,很可能参与了 HIV/AIDS患者粘膜免疫系统损伤过程,应在HIV/AIDS粘膜损伤修复中予以重视。
[Abstract]:Aim: polymeric immunoglobulin receptor pIgR, an important component of mucosal immune system, plays a key role not only in the transport of IgA from the intrinsic membrane to the mucosal surface, but also directly in the composition of SIgA. Therefore, the effective secretion of pIgR is a necessary condition for polyimmunoglobulin D IgA and pIgM to play a normal mucosal defense function. Although SIgA is seriously deficient in HIV/AIDS patients, it is not clear that the pIgR expression level and its distribution after HIV / IV infection are unknown. In this study, the expression and distribution of pIgR in intestinal and respiratory mucosa of rhesus monkeys after SIV/SHIV infection and the effect of IL-17A on pIgR expression in cultured human bronchial epithelial cells were observed. To elucidate the changes and possible mechanisms of intestinal and respiratory pIgR after HIV infection. To provide the necessary scientific basis for the reconstruction of mucosal immune system in diseases related to mucosal injury such as AIDS. Methods: the expression of pIgRmRNA and its expression in normal and SIV/SHIV infected rhesus monkey mucosa were detected by routine and real-time fluorescence quantitative RT-PCR. Western blotting was used to detect the expression of pIgR protein in normal and infected rhesus monkey mucosa. The distribution and abundance of pIgR protein in normal and infected rhesus monkey mucosa were observed by immunofluorescence labeling and laser scanning confocal microscopy. In addition, the effects of IL-17A on the levels of pIgR mRNA and pIgR protein in human bronchial epithelial cells were observed by cell culture in vitro. Results: the expression of pIgR protein in stomach, duodenum, jejunum, ileum, cecum, colon and rectum of normal rhesus monkey were different. The expression of pIgR protein decreased significantly in the corresponding tissues after SIV/SHIV infection. In the gastric fundus, proximal jejunum, cecum, colon and rectum, the difference was statistically significant (P = 0.033). The expression of pIgRmRNA and pIgR protein in tracheal mucosa decreased significantly after infection (P = 0.0014) IL-17 mRNA expression was also decreased after SIV/SHIV infection. There was a positive correlation between pIgR mRNA and IL-17A mRNAs. The expression of pIgR mRNA and IL-17AmRNA in lung tissues increased after infection. There was a positive correlation (positive correlation, but the expression of pIgR protein decreased after infection. 3) the relative expression of pIgR mRNA increased first and then decreased within 72 h after HBE stimulated by different concentrations of IL-17A, and a maximum value appeared within 24 h. Conclusion: these results suggest that pIgR transcription and protein expression in the digestive tract mucosa and tracheal mucosa of rhesus monkey infected with SIV / SHIV can be decreased to some extent. IL-17A can promote the expression of pIgR in mucosal epithelial cells to some extent. The abnormal expression of pIgR may be related to the decrease of IL-17A level, which may be involved in the process of mucosal immune system injury in HIV/AIDS patients. It should be paid more attention to in the repair of HIV/AIDS mucosal damage.
【学位授予单位】：中国疾病预防控制中心
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R512.91

【参考文献】

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1 Dong Li;Feng-Jie Wang;Lei Yu;Wen-Rong Yao;Yan-Fang Cui;Gui-Bo Yang;;Expression of pIgR in the tracheal mucosa of SHIV/SIV-infected rhesus macaques[J];Zoological Research;2017年01期

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本文编号：1787626