Elucidation of Host and Viral factors in HIV-1 infected individuals.

Abstract

In the last three decades, the world has known HIV/AIDS as a disease that has been responsible for one of the major anthropological epidemics our globe has faced since the time it was discovered. From the time HIV/AIDS was initially made known in 1982 and HIV revealed in 1983, marvelous advancement has been achieved in knowing the source of its evolutionary tree as well as knowing the fundamental biology of the this virus, the intricacies of its communication with the complex immune system of humans, and the various ways accountable for its spread, early propagation, and pathogenesis.. The role of host factors such as genetic single nucleotide polymorphisms’ (SNPs) and the viral sequences in the variance seen in susceptibility to HIV infection, viral spread, immune response to therapeutic interventions and the pace of progression to AIDS is established in other populations. However, no such study is available from Pakistani population. For this purpose, three cytokines IL-10, TGF-β and IL-18 were chosen for investigating their association with HIV pathogenesis in Pakistani patients. The viral integrase and part of RNAse H gene was also sequenced. A possible association was sought for 40 HIV-1 patients and controls for IL-18 -607C/A, IL-10-1082A/G, and TGF-509C/T polymorphisms. The SNPs were verified using allele specific PCR as well as restriction fragment length polymorphism analysis (RFLP). A 161 and 700 amplicon within the integrase gene was also optimized using in-house PCR and thereafter sequenced. The study found that there was no significant difference amongst the HIV patients and control for polymorphisms in IL-10 and IL-18, however higher IL-10-GG genotypes were found in the control group which xii is linked to faster progression to AIDS as well as higher IL-18-CC/CA genotypes amongst both the groups which is also linked to rapid progression to AIDS. Moreover, a higher TGF-β -509TT genotype and T allele was found in half of the HIV patients suggesting its strong association with a high viral load. The 161bp and 700bp amplicons were successfully amplified from all HIV positive patients for designing of a future in-house HIV-1 detection assay as well as few 700bp amplicons were sequenced for future phylogenetic analysis. These results may aid in future cohort studies as well as genotyping, diagnostics and drug resistance mutation analysis.