DTG 3TC HIV EPI study.xlsx

The persistence of human immunodeficiency virus (HIV) within viral reservoirs poses significant challenges to eradication efforts. Epigenetic alterations, including DNA methylation, are potential factors influencing HIV latency and persistence. This study details the development and application of techniques to assess CpG methylation in the promoter regions of the <i>CCR5</i> and <i>CXCR4</i> genes, key HIV-1 coreceptors. Using both Sanger sequencing and pyrosequencing methods, we examined 51 biological samples from 17 HIV-1-infected individuals at three time points: baseline (Week 0) and post-antiretroviral therapy (ART) at Weeks 24 and 48.Our results revealed that <i>CXCR4</i> promoter CpG sites were largely unmethylated, while <i>CCR5</i> promoter CpGs exhibited significant variability in methylation levels. Specifically, <i>CCR5</i> CpG 1 showed a significant increase in methylation from Week 0 to Week 48, while <i>CXCR4</i> CpG 3 displayed a significant decrease between Week 0 and Week 24. These differences were statistically significant when compared with non-HIV-infected controls. These findings demonstrate distinct methylation patterns between <i>CCR5</i> and <i>CXCR4 </i>promoters in HIV-1 individuals over time, suggesting that epigenetic modifications may play a role in regulating HIV-1 persistence. Our techniques provide a reliable framework for assessing gene promoter methylation and could be applied to further research in HIV epigenetics.<br>