IL-27及其受体在艾滋病发病机制和抗病毒免疫反应中的作用

发布时间：2018-04-28 15:41

本文选题：IL-27 + gp130/CD130　；参考：《中南大学》2014年硕士论文

【摘要】：背景：艾滋病(Acquired Immune Deficiency Syndrome, AIDS)是严重威胁人民健康的公共卫生问题,随着高效抗反转录病毒治疗(Highly active antiretro viral therapy, HAART)的推广,艾滋病抗病毒治疗取得了一定的成效,但迄今为止仍无有效的方法彻底清除艾滋病毒(human immunodeficiency virus, HIV)。近年来,细胞因子在AIDS的发病过程及其抗病毒治疗中的作用受到日益关注。科学家们已发现了多种细胞因子参与抗HIV病毒免疫应答和免疫重建,并提出其中一些细胞因子可能成为加强抗HIV病毒效果的辅助治疗手段。IL-27是HIV研究领域一个较新的细胞因子,已有体外实验发现IL-27在HIV-1病毒感染中可能存在一定的治疗价值,但IL-27相关的临床研究较少。本文将研究HIV感染者IL-27及其受体的表达情况及其在艾滋病发病机制和HAART抗病毒治疗免疫反应中的作用。目的：研究HIV感染者IL-27和它的受体表达情况及其与正常人的区别；HAART对IL-27和它的受体表达的影响；分析HIV感染者IL-27及其受体gp130与CD4+T细胞数、HIV-RNA病毒载量的关系,IL-27与Thl细胞因子IFN-y和Th2细胞因子IL-4的关系,进一步探索艾滋病的发病机制和免疫重建机制。方法：从2012年12月到2013年3月来湘雅二医院艾滋病专科门诊就诊并准备启动HAART治疗的HIV感染者/AIDS患者中选取34例纳入本研究,并定期随访收集HAART治疗开始前(0月),开始治疗6月、12月的血标本。同时随机选取健康体检者27例作为健康对照组。使用酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)检测血浆中IL-27、IFN-γ、IL-4的浓度,流式细胞术检测外周血CD3+CD4+T细胞数、外周血单个核细胞(Peripheral Blood Mononuclear Cell, PBMC)中CD4+细胞表面gp130的表达情况,HIV实时荧光定量PCR测定外周血中HIV-RNA病毒载量,RT-PCR和琼脂糖凝胶电泳检测gp130mRNA的表达。对以上所测得数据进行统计学处理和相关性分析。结果:HIV感染未治疗者血浆中IL-27的平均浓度明显低于正常人,开始HAART治疗后IL-27的浓度逐渐升高,治疗6月及12月后血浆中IL-27浓度明显高于未治疗前。HIV感染者PBMC中CD4+细胞表面gp130表达水平明显低于正常人,随着HAART抗病毒治疗的进行,可逆转这种情况,使gp130的表达逐渐增加；治疗6月、12月时CD4+细胞表面gp130表达水平明显高于未治疗前。通过RT-PCR、琼脂糖凝胶电泳半定量检测gp130mRNA的表达水平证实了细胞表面gp130表达趋势的变化。IL-27浓度与CD4+细胞表面gp130表达水平成正相关；IL-27和它的受体gp130均与CD4+T细胞数成正相关、与血浆HIV-RNA病毒载量成负相关。HIV感染者血浆IFN-y浓度随着治疗时间的延长越来越高,IL-4的浓度随着治疗时间的延长越来越低,其浓度水平都在向正常人群靠近。在HIV感染者中IL-27浓度的变化趋势与IFN-y一致,与IL-4相反。结论： 1、首次观察到HIV病毒感染者血浆IL-27浓度低于健康人群,且随着HAART抗病毒治疗,IL-27的浓度逐渐升高；同时观察到HIV病毒感染者IL-27的受体gp130在CD4+T细胞表面的表达水平明显低于健康人群,随着HAART抗病毒治疗时间的延长,这种受体表达受损可以逐渐恢复。 2、发现HIV病毒感染者外周学IL-27的浓度及其受体gp130的表达与血浆HIV-RNA水平成负相关,与CD3+CD4+T细胞数量成正相关。提示血浆中的IL-27对HIV感染者可能具有保护效应,IL-27及其受体gp130可能参与了HIV的发病机制及抗病毒免疫反应机制。 3、IL-27的浓度变化和IFN-y与IL-4的浓度变化存在一定的关系,提示IL-27在HIV病毒感染中以炎性作用为主,且可能与Thl/Th2细胞因子失衡有关。
[Abstract]:Background: Acquired Immune Deficiency Syndrome (AIDS) is a public health problem that seriously threatens the people's health. With the promotion of high performance antiretroviral therapy (Highly active antiretro viral therapy, HAART), AIDS antiviral treatment has achieved a definite effect, but so far there are still no effective methods to eliminate it. Human immunodeficiency virus (HIV). In recent years, the role of cytokines in the pathogenesis of AIDS and its antiviral therapy has been increasingly concerned. Scientists have discovered a variety of cytokines involved in the anti HIV virus immune response and immune reconstruction, and suggest that some of its cytokines may become the strengthening of the anti HIV virus. .IL-27 is a new cytokine in the research field of HIV. It has been found that IL-27 may have some therapeutic value in HIV-1 virus infection in vitro, but there are few clinical studies on IL-27. This paper will study the expression of IL-27 and its receptor in HIV infected persons and the pathogenesis of AIDS and HAART resistance. The role of virus in the treatment of immune response.
Objective: To study the expression of IL-27 and its receptor and the difference between HIV and normal people, the effect of HAART on the expression of IL-27 and its receptor, the relationship between the number of IL-27 and its receptor gp130 and the number of CD4+T cells, the number of HIV-RNA virus, the relationship between IL-27 and Thl cytokine IFN-y and cytokines, and further explore the relationship between the IL-27 and Thl cytokine IFN-y and cytokines. The pathogenesis of AIDS and the mechanism of immune reconstruction.
Methods: from December 2012 to March 2013, 34 cases of /AIDS patients with HIV infection in the AIDS specialist clinic in Xiangya No.2 Hospital and /AIDS patients prepared to start the treatment were selected to be included in this study, and the regular follow-up collection of HAART treatment began (0 months) and began to treat the blood scale in June and December. At the same time, 27 health examiners were randomly selected as health. In the control group, enzyme-linked immunosorbent assay (ELISA) was used to detect the concentration of IL-27, IFN- gamma, and IL-4 in plasma. Flow cytometry was used to detect the number of CD3+CD4+T cells in peripheral blood, and the expression of peripheral blood mononuclear cells (Peripheral Blood Mononuclear Cell). The HIV-RNA virus load in peripheral blood was measured by CR, and the expression of gp130mRNA was detected by RT-PCR and agarose gel electrophoresis. Statistics and correlation analysis were carried out for the above data.
Results: the average concentration of IL-27 in plasma of HIV infected people was significantly lower than that of normal people, and the concentration of IL-27 increased gradually after HAART treatment. The concentration of IL-27 in plasma was significantly higher than that of.HIV infected persons in June and December. The level of gp130 expression on the surface of CD4+ cells in PBMC was significantly lower than that of normal people, with the antiviral therapy of HAART. It can reverse this situation and increase the expression of gp130 gradually. The expression level of gp130 on the surface of CD4+ cells in June and December is obviously higher than that before the treatment. The expression level of gp130mRNA by RT-PCR and agarose gel electrophoresis confirms the gp130 expression trend of the cell surface and the gp130 expression on the surface of CD4+ cells. The level of IL-27 and its receptor gp130 were positively correlated with the number of CD4+T cells. The concentration of IFN-y in the plasma of.HIV infected with HIV-RNA virus was increasing with the prolongation of the treatment time. The concentration of IL-4 was decreasing with the time of treatment, and the concentration level was close to the normal population. In HIV infection, the concentration level of the IL-4 was in the HIV infection. The change trend of IL-27 concentration was consistent with that of IFN-y, contrary to IL-4.
Conclusion:
1, the concentration of IL-27 in plasma of HIV virus infected people was lower than that of healthy people for the first time, and with HAART antiviral therapy, the concentration of IL-27 increased gradually. Meanwhile, the expression level of IL-27 receptor gp130 on the surface of CD4+T cells was obviously lower than that of the healthy population, and this receptor table was prolonged with the prolongation of the time of HAART antiviral therapy. The damage can be recovered gradually.
2, it was found that the concentration of IL-27 in peripheral study and the expression of gp130 and the expression of gp130 were negatively related to the plasma HIV-RNA level, and were positively related to the number of CD3+CD4+T cells. It suggests that IL-27 in plasma may have protective effect on HIV infected persons. IL-27 and its receptor gp130 may be involved in the pathogenesis of HIV and the mechanism of antiviral immune response.
3, the concentration changes of IL-27 and the changes of the concentration of IFN-y and IL-4 have a certain relationship, suggesting that IL-27 is mainly inflammatory in HIV virus infection, and may be related to the imbalance of Thl/Th2 cytokine.

【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R512.91

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