Hepatitis C virus genetic variability in 52 human immunodeficiency virus‐coinfected patients

Abstract

The aim of this study was to examine whether hepatitis C virus (HCV) pretreatment quasispecies complexity was linked to virological response or other clinical and biological parameters, in human immunodeficiency virus (HIV)-coinfected patients undergoing anti-HCV treatment. In addition, HCV quasispecies composition is described longitudinally in these patients before, during, and after treatment. The 52 HIV-coinfected patients were included in a randomized therapeutic trial. At inclusion, they had CD4(+) counts of >250/micro l, HIV plasma load of <10,000 copies/ml, and chronic HCV infection with genotype 1 (n = 27), 2 (n = 2) or 3 (n = 23). These values were compared at baseline with 32 HCV-only-infected, interferon-naive patients who were infected with genotype 1, 2, or 3 (n = 16, 1, or 15, respectively). HCV complexity was studied by single-strand conformation polymorphism (SSCP) in E2 hypervariable region 1 (HVR1), and diversity was evaluated at inclusion in 20 coinfected patients by sequencing four major SSCP bands. The baseline number of SSCP bands was identical in HIV-infected and control patients. In HIV-infected patients, HCV complexity was not predictive of sustained virological response to anti-HCV treatment and was unrelated to epidemiological factors, immunological parameters linked to HIV infection (CD4(+) counts, T CD4(+) proliferative responses to HIV-1 p24), protease inhibitor treatment, HCV plasma load, or genotype. HCV diversity was lower in genotype 2- and 3-infected patients. Six months after completion of the anti-HCV treatment, in comparison with baseline, SSCP profiles were modified in 13 of the 21 nonresponding coinfected patients with analyzable samples. In conclusion, in HIV-infected patients, HCV variability had no significant influence on virological response to anti-HCV treatment.

HIV Infection and Antiretroviral Therapy: Effect on Hepatitis C Virus Quasispecies Variability

“…A third study reported a trend toward decreased complexity in 27 HCV/HIV-coinfected subjects, compared with 10 subjects infected with HCV alone, although it found no difference in diversity between the 2 groups [24]. The largest study to date compared 52 HCV/HIV-coinfected patients who were entering an HCV treatment trial with 32 HIV-negative untreated control subjects, and it found that quasispecies complexity was identical in the 2 groups [40]. However, HIV treatment status was not well defined, and quasispecies analysis was performed using the single-strand conformational polymorphism technique, which is the least accurate method for analyzing complex genetic populations, because genetically different variants frequently comigrate during SSCP and are impossible to resolve.…”

“…In light of the aforementioned controversy, we studied a relatively large number of well-characterized subjects and carefully controlled for variables that have been associated with quasispecies variability, including duration of HCV infection [41], genotype [25,28,40], HCV load [28], alcohol use [42,43], and ART [27,28]. Although the present study is limited by its crosssectional design, we decided a priori to exclude subjects who might be in various stages of immune reconstitution, to have immunologically and clinically distinct groups.…”

Early Evolution of Hepatitis C Virus (HCV) Quasispecies after Liver Transplant for HCV‐Related Disease

“…Extraction of total RNA was performed in plasma (QIAmp Viral RNA Mini Kit; Qiagen). The same starting amount of HCV RNA was always used next to a 250-300-ng reversetranscription polymerase chain reaction (PCR), which was performed on HVR1 using published primers [6] and a Proofstart DNA polymerase (Qiagen). PCR products were cloned by using the TOPO TA Cloning kit (Invitrogen) in accordance with the manufacturer's recommendations (pCR II-TOPO vector, competent Escherichia coli TOP10).…”

Plasma and Liver Hepatitis C Virus Variability in Patients Coinfected with Human Immunodeficiency Virus

“…After RNA extraction from plasma and liver (22), a 325-bp fragment encompassing HVR1 region was amplified (11) and cloned (pGEM-T Easy Vector System I; Promega). For the three patients coinfected with HIV 222 clones were evaluated (mean, 18.5 per sample), and for the three HIV-negative patients 95 clones were evaluated.…”