The persistence of human immunodeficiency virus (HIV) in viral reservoirs is a topical issue. There are many factors that can potentially influence this process. Epigenetic alterations according to the literature could be such factors. In this study, we present a detailed description of gene promoters research and development techniques for determining the methylation level of CpG loci in these promoters based on Sanger sequencing and pyrosequencing. The presented methods could be used for measuring the methylation levels in other research.
A total of 51 samples of biological material from 17 patients at three visits were used in this study. Applying the developed techniques, we found that the CpG loci of theCXCR4gene promoter were not methylated. For CpG loci of theCCR5gene promoter no change in methylation levels was observed visit to visit. CpG 1 methylation level ranged from 33% to 53%, CpG 2 - from 4.4% to 11%, CpG 3 - from 0 to 42% and CpG 4 - from 1% to 21%. These values were derived from measurements of C-peaks in sequences.
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