The Correlation Between The Status Of T Lymphocyte And HLA Genotype Of HIV-1 Highly Exposed Persistently Seronegative Individuals (ESNs)

Objective1. Setting HIV-1-exposed seronegative subjects (ESNs) of Han Chinese as the research objective, healthy people and Human Immunodeficiency Virus Type 1(HIV-1) carriers as the control population, to explore the relationship between the gene polymorphism of DC-SIGN, DC-SIGNR and HIV-1 low susceptibility.2. To analyzing the expression difference of HIV-1 infection related co-receptor, from the level of protein expression to search the co-receptor related with HIV-1 low susceptibility. 3. To clarify the reason of expression difference of co-receptor from the view of gene polymorphism.MethodsPCR-ALFP method was applied to analyze DC-SIGN, DC-SIGNR gene polymorphism. Flow cytometer was used to analyze the expression difference of HIV-1 infection related co-receptor, PCR-SSP was used to analyze HLA gene polymorphism, SPSS 13.0 was applied to analyze the difference of co-receptor expression and genotype distribution.Results1. Genotype 7/7 was the primary genotype of DC-SIGN. Only 9 non 7/7 genotype was tested in three groups. There was no significant difference between 7/7 and non-7/7 genotypes(P=0.648).DC-SIGNR genetic polymorphism was high, and the frequency of 5/5 and 6/7 genotypes among ESN, healthy control and HIV-1 carriers gradually decrease. The allele frequency of 5 and 6 also had the same trend. But there were no significant difference in genotypes (P=0.782) and gene alleles between three groups.2. We found the ESNs had significantly lower percentage of HLA-DR+ CD4 T cells and HLA-DR+ CD8 T cells than healthy controls and HIV-1 carriers (P=0.024,0.009). The two index also had significant positive correlation (P<0.001, r=0.960). Healthy controls could be clearly divided into low expression group and high expression group according to HLA-DR+ CD8 T cells expression.3. According to Fisher's exact Chi-square test, only A*03 and B*55 genotype distribution had statistic significance. In healthy controls, A*02 and A*24 genotype distribution shown significant difference between HLA-DR+CD8 low and high expression groups (P=0.020, 0.038). The analysis of allele distribution also confirm above results of the difference between the two gene polymorphism (P=0.007,0.009). DRB1*09 also show significant difference in allele distribution. The distribution of A*02 allele showed significant negative correlation with HLA-DR+CD8 expression (P=0.008), whereas the distribution of A*24 displayed a significant positive correlation with HLA-DR+CD8 expression (P=0.045).Conclusions1. Han Chinese has very low mutation frequency in DC-SIGN. It is not necessary to analyze DC-SIGN polymorphism in future study. We need large sample or data of cell biology to conform the relation between HIV-1 susceptibility and DC-SIGNR polymorphism.2. To Han Chinese, the low activation status of T lymphocyte has close correlation with HIV-1 low susceptibility. Han Chinese can be divided into low and high susceptibility groups according to activation status of HLA-DR+CD8 from the aspects of statistic analysis and biological significance.3. HLA-A*02 polymorphism mutation may be the protect factor of HIV-1 infection and HLA-A*24 polymorphism mutation may be the risk factor of HIV-1 infection to Han Chinese. Innovative Point1. Firstly set quantitive enrollment standard in ESNs research in China.2. Firstly report the close relationship between HLA-DR high/low expression and HIV-1 susceptibility of Han Chinese.3. Firstly report the result that HLA-A*02 polymorphism mutation may be the protect factor of HIV-1 infection and HLA-A*24 polymorphism mutation may be the risk factor of HIV-1 infection to Han Chinese.