Hepatitis C Virus (HCV) NS3 Sequence Diversity and Antiviral Resistance-Associated Variant Frequency in HCV/HIV Coinfection

HIV coinfectionaccelerates disease progression in chronic hepatitis C and reduces sustained antiviral responses (SVR) to interferon-based therapy. New direct-acting antivirals (DAAs) promise higher SVR rates, but the selection of preexisting resistance-associated variants (RAVs) may lead to virologic breakthrough or relapse. Thus, pretreatment frequencies of RAVs are likely determinants of treatment outcome but typically are below levels at which the viral sequence can be accurately resolved. Moreover, it is not known how HIV coinfectioninfluences RAV frequency. We adopted an accurate high-throughput sequencing strategy to compare nucleotide diversityin HCV NS3protease-coding sequences in 20 monoinfected and 20 coinfectedsubjects with well-controlledHIV infection. Differences in mean pairwise nucleotide diversity(π), Tajima's Dstatistic, and Shannon entropy index suggested that the genetic diversity of HCV is reduced in coinfection. Among coinfectedsubjects, diversity correlated positively with increases in CD4+ T cells on antiretroviral therapy, suggesting T cell responses are important determinants of diversity. At a median sequencing depth of 0.084%, preexisting RAVs were readily identified. Q80K, which negatively impacts clinical responses to simeprevir, was encoded by more than 99% of viral RNAs in 17 of the 40 subjects. RAVs other than Q80K were identified in 39 of 40 subjects, mostly at frequencies near 0.1%. RAV frequency did not differ significantly between monoinfected and coinfectedsubjects. We conclude that HCV genetic diversity is reduced in patients with well-controlledHIV infection, likely reflecting impaired T cell immunity. However, RAV frequency is not increased and should not adversely influence the outcome of DAA therapy.