HIV-1高暴露未感染人群淋巴细胞活化状态与HLA分型的关系

发布时间：2018-03-20 22:51

本文选题：HIV-1　切入点：ESNs人群　出处：《华中科技大学》2011年博士论文　论文类型：学位论文

【摘要】：研究目的 1.以中国汉族HIV-1暴露而血清阴性者(HIV-1-exposed seronegative subjects, ESNs人群)为研究对象,以健康对照人群和人类免疫缺陷病毒I型(Human Immunodeficiency Virus Type 1, HIV-1)携带者为对照,探讨三组人群DC-SIGN和DC-SIGNR基因多态性与HIV-1不易感性的关系。 2.分析三组人群HIV-1感染相关受体的表达差异,从蛋白质表达水平上寻找与HIV-1不易感性相关的受体。 3.从基因多态性的角度解释与HIV-1不易感性密切相关的受体差异表达原因。研究方法应用扩增片段长度多态性(PCR-ALFP)的方法进行DC-SIGN和DC-SIGNR基因多态性分析；应用流式细胞术分析HIV-1感染相关受体的表达差异；应用序列特异引物PCR (sequence-specific primers, PCR-SSP)分析HLA基因多态性；应用SPSS13.0统计软件分析三组人群之间受体表达和基因型分布的差异。主要研究结果 1. DC-SIGN在三组人群中以7/7型为主,三组人群共检出9个非7/7基因型,但DC-SIGN各基因型分布差异无统计学意义(P=0.648); DC-SIGNR具有高度多态性,7/5、5/5和6/7基因型的频率从ESN人群、健康对照人群和HIV-1携带者逐渐降低,同时5和6等位基因的频率也呈现均匀下降,但各基因型分布差异无统计学意义(P=0.782)； 2. ESNs人群T淋巴细胞HLA-DR+CD4和HLA-DR+CD8均显著性低于健康对照人群(P=0.024,0.009),这两项指标同时呈现显著性正相关(P0.001,r=0.960)；利用HLA-DR+CD8的双峰分布特征可以将健康对照人群分为高低表达两群； 3.确切概率法卡方检验结果显示,三个基因座的多态性位点中只有A*03和B*55在三组人群中的分布差异有显著性。健康对照人群HLA基因型的分析发现,A*02和A*24基因型在HLA-DR+CD8表达高低两组的分布有显著性差异(P=0.020,0.038)；等位基因分析也验证了这个多态性变异分布的差异(P=0.007,0.009),DRB1*09等位基因分布也有显著性差异(P=0.028)；A*02多态性变异与HLA-DR在CD8阳性细胞表面的表达有显著性负相关(P=0.008)；A*24多态性变异与HLA-DR在CD8阳性细胞表而的表达有显著性正相关(P=0.045)。研究结论 1.我国汉族人群DC-SIGN基因多态性变异很低,没有继续研究的意义；DC-SIGNR基因多态性对于HIV-1不易感性的意义需要大样本的验证或细胞生物学实验数据的支持； 2.对于中国汉族人群,T淋巴细胞低活化状态与中国汉族人群HIV-1不易感性密切相关,从统计学分析和生物学意义上可以将中国汉族人群按照HLA-DR+CD8活化高低分为HIV-1易感人群和不易感人群； 3. HLAA*02多态性变异可能是中国汉族人群HIV-1感染的保护性因素,而HLAA*24多态性变异可能是感染的危险因素。创新性 1.首次在国内ESNs人群的研究中设定了定量的的入选标准； 2.在国内首次报道HLA-DR的高低表达与中国汉族人群HIV-1不易感性密切相关； 3.首次报道HLA-A*02多态性变异可能是中国汉族人群HIV-1感染的保护性因素,而HLA-A*24多态性变异可能是感染的危险因素。
[Abstract]:research objective
1. to Chinese Han HIV-1 exposed seronegative individuals (HIV-1-exposed seronegative subjects, ESNs group) as the research object, to a group of healthy people and human immunodeficiency virus type I (Human Immunodeficiency Virus Type 1, HIV-1) carriers as control, three groups of DC-SIGN and DC-SIGNR gene polymorphism and HIV-1 susceptibility.
2. the differences in the expression of HIV-1 related receptors in the three groups were analyzed, and the receptors associated with the HIV-1 non susceptibility were found from the protein expression level.
3. from the perspective of genetic polymorphisms, the reasons for the differential expression of the receptor, which is closely related to the HIV-1 susceptibility, are explained.
research method
Application of amplified fragment length polymorphism (PCR-ALFP) method for the analysis of DC-SIGN and DC-SIGNR gene polymorphism; analysis of differentially expressed HIV-1 infection related receptors by flow cytometry; using sequence specific primer PCR (sequence-specific primers PCR-SSP) analysis of HLA gene polymorphism; analysis of receptor expression and genotype differences between the three groups people using SPSS13.0 statistical software.
Main research results
1. DC-SIGN to 7/7 in the three groups, the three groups were detected in 9 non 7/7 genotype, but the genotype distribution of DC-SIGN showed no significant difference (P=0.648); DC-SIGNR has highly polymorphism, 7/5,5/5 and 6/7 genotypes from ESN group, healthy control group and HIV-1 carriers decreased gradually at the same time, the 5 and 6 allele frequencies showed uniform decline, but the genotype distribution difference was not statistically significant (P=0.782);
2. ESNs HLA-DR+CD4 and HLA-DR+CD8 T lymphocyte populations were significantly lower than those in healthy controls (P=0.024,0.009), these two indicators also showed significant positive correlation (P0.001, r=0.960); the Shuangfeng HLA-DR+CD8 distribution can be divided into healthy control group two group level expression;
3. exact test chi square test showed that there was significant difference in the distribution of only A*03 and B*55 in the three groups in the three polymorphic loci loci. Analysis of healthy control group HLA genotype, A*02 genotype distribution and A*24 level of the two groups had significant difference in the expression of HLA-DR+CD8 (P=0.020,0.038); allele analysis also verified the polymorphism distribution difference (P=0.007,0.009), DRB1*09 allele distribution had significant difference (P=0.028); A*02 polymorphism and HLA-DR has significant negative correlation in the expression of CD8 positive cell surface (P=0.008); A*24 polymorphism and HLA-DR in CD8 positive cells and expression have significant positive correlation (P=0.045).
research conclusion
1., the variation of DC-SIGN gene polymorphism in Chinese Han population is very low, which has no significance for further research. The significance of DC-SIGNR gene polymorphism for HIV-1 is not susceptible, which needs large sample validation or cell biology experimental data support.
2., for Chinese Han population, the low activation status of T lymphocytes is closely related to HIV-1's susceptibility to Chinese Han population. From statistical analysis and biological significance, Chinese Han population can be divided into HIV-1 susceptible group and non susceptible group according to HLA-DR+CD8 activation level.
3. HLAA*02 polymorphism may be a protective factor for HIV-1 infection in Chinese Han population, and HLAA*24 polymorphism may be a risk factor for infection.
Innovativeness
1. for the first time, a quantitative admission standard was set in the study of the domestic ESNs population.
2. the high and low expression of HLA-DR is closely related to the non susceptibility of HIV-1 in Chinese Han population for the first time in China.
3., for the first time, it is reported that HLA-A*02 polymorphism may be a protective factor of HIV-1 infection in Chinese Han population, while HLA-A*24 polymorphism may be a risk factor for infection.

【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2011
【分类号】：R181.3

【引证文献】