發(fā)布時間：2018-04-22 14:23

  本文選題：多聚免疫球蛋白受體 + SIV/SHIV感染��； 參考：《中國疾病預防控制中心》2017年碩士論文

【摘要】：目的:多聚免疫球蛋白受體(polymeric immunoglobulin receptor,pIgR)是黏膜免疫系統(tǒng)的重要組成成分,不僅在IgA從固有膜到粘膜表面的轉(zhuǎn)運過程中發(fā)揮關(guān)鍵作用,而且直接參與SIgA的構(gòu)成。因此,pIgR的有效分泌是多聚免疫球蛋白(dIgA和pIgM)發(fā)揮正常粘膜防御功能的必要條件。雖然SIgA在HIV/AIDS患者中出現(xiàn)嚴重缺陷,但對pIgR表達水平與分布在免疫缺陷病毒(HIV/SIV)感染后的變化規(guī)律尚不清楚。本研究對恒河猴腸道和呼吸道粘膜中pIgR表達水平與分布在SIV/SHIV感染后的變化以及IL-17A對體外培養(yǎng)人支氣管上皮細胞中pIgR表達的影響進行了觀察,擬闡明腸道和呼吸道pIgR在免疫缺陷病毒感染后的變化規(guī)律及可能機制。為艾滋病等黏膜損傷相關(guān)疾病中粘膜免疫系統(tǒng)的重建提供必要的科學基礎(chǔ)。方法:本研究利用常規(guī)和實時熒光定量RT-PCR方法檢測正常和SIV/SHIV感染恒河猴粘膜組織中pIgRmRNA及其表達水平。利用Western blotting檢測正常和感染恒河猴粘膜組織中pIgR蛋白及其表達水平。利用免疫熒光標記及激光掃描共聚焦顯微鏡觀察正常和感染恒河猴粘膜組織中pIgR蛋白的分布和豐度。另外,利用體外細胞培養(yǎng)技術(shù)觀察IL-17A對人支氣管上皮細胞(HBE)中pIgR mRNA水平和pIgR蛋白水平的影響。結(jié)果:研究發(fā)現(xiàn):1)正常恒河猴胃底、十二指腸、空腸、回腸、盲腸、結(jié)腸和直腸pIgR蛋白表達水平存在部位間差異,小腸和大腸的前端表達水平較高。在SIV/SHIV感染后相對應(yīng)的組織中pIgR蛋白表達水平明顯下降。在胃底、空腸近端、盲腸、結(jié)腸和直腸中,差異具有統(tǒng)計學意義,P值分別為 0.033,0.0043,0.0364,0.0252,0.0424。2)氣管粘膜中pIgRmRNA和pIgR蛋白表達水平在感染后均顯著下降,P值為0.0014;IL-17AmRNA表達水平在SIV/SHIV感染后也下降,并且兩者(pIgR mRNA vs IL-17A mRNA)具有相關(guān)關(guān)系(正相關(guān))。在肺組織中pIgR mRNA和IL-17AmRNA表達水平在感染后均有所上升,兩者具有相關(guān)性(正相關(guān)),但pIgR蛋白表達水平在感染后下降。3)不同濃度IL-17A刺激HBE以后,pIgR mRNA的相對表達量在72 h內(nèi)均呈現(xiàn)先增加后減少的趨勢,在24h內(nèi)有一個最高值出現(xiàn)。結(jié)論:上述結(jié)果表明,SIV/SHIV感染恒河猴消化道粘膜和氣管粘膜中pIgR轉(zhuǎn)錄水平和蛋白表達水平都出現(xiàn)不同程度的降低,IL-17A對粘膜上皮細胞pIgR的表達具有一定的促進作用。免疫缺陷病毒感染后粘膜pIgR表達異�？赡芘cIL-17A水平下降有關(guān),很可能參與了 HIV/AIDS患者粘膜免疫系統(tǒng)損傷過程,應(yīng)在HIV/AIDS粘膜損傷修復中予以重視。
[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.
【學位授予單位】：中國疾病預防控制中心
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R512.91

【參考文獻】

相關(guān)期刊論文 前1條

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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本文編號：1787626