RESUMEN
OBJECTIVE: replication of HIV-1 after cell entry is essentially dependent on the reverse transcriptase (RT). Antiretroviral drugs impairing the function of the RT currently aim at the polymerase subunit. One reason for failure of antiretroviral treatment is the evolvement of resistance-associated mutations in the viral genome. For RT inhibitors, almost all identified mutations are located within the polymerase; therefore, general genotyping confines to investigate this subunit. Recently several studies have shown that substitutions within the RNase H and the connection domain increase antiviral drug-resistance in vitro, and some of them are present in patient isolates. AIM: the aim of the present study was to investigate the prevalence of these substitutions and their association with mutations in the polymerase domain arising during antiretroviral treatment. MATERIAL AND METHODS: we performed genotypic analyzes on seventy-four virus isolates derived from treated and untreated patients, followed at the HIV Centre of the Johann Wolfgang Goethe University Hospital (Frankfurt/Main, Germany). We subsequently ana?lysed the different substitutions in the c-terminal region to evaluate whether there were associations with each other, n-terminal substitutions or with antiretroviral treatment. RESULTS: We identified several primer grip substitutions, but almost all of them were located in the connection domain. This is consistent with other in-vivo studies, in which especially the primer grip residues located in the RNase H were unvaried. Furthermore, we identified other substitutions in the connection domain and in the RNase H. Especially E399D seemed to be associated with an antiretroviral treatment and N-terminal resistance-delivering mutations. CONCLUSION: some of the identified substitutions were associated with antiviral treatment and drug resistance-associated mutations. Due to the low prevalence of C-terminal mutations and as only a few of them could be associated with antiviral treatment and N-terminal resistance-delivering mutations, we would not recommend routinely testing of the C-terminal RT region.
Asunto(s)
Fármacos Anti-VIH/uso terapéutico , Transcriptasa Inversa del VIH/genética , VIH-1/efectos de los fármacos , Mutación , Sustitución de Aminoácidos , Antirretrovirales/uso terapéutico , Resistencia a Medicamentos , Infecciones por VIH/virología , VIH-1/enzimología , VIH-1/genética , Humanos , Reacción en Cadena de la Polimerasa/métodos , ARN Viral/genética , Polimerasa Taq/genética , Replicación Viral/efectos de los fármacos , Zidovudina/uso terapéuticoRESUMEN
The evolution of intra-host human immunodeficiency virus type 1 (HIV-1) quasispecies prior and after treating active tuberculosis (TB) with chemotherapy in HIV-1/TB patients was assessed. Two time points HIV-1 quasispecies were evaluated by comparing HIV-1-infected patients with active tuberculosis (HIV-1/TB) and HIV-1-infected patients without tuberculosis (HIV-1/non-TB). Plasma samples were obtained from the Frankfurt HIV cohort, and HIV-1 RNA was isolated. C2V5 env was amplified by PCR and molecular cloning was performed. Eight to twenty-five clones were sequenced from each patient. Various phylogenetic analyses were performed. We found a significant increase in diversity and divergence in HIV-1/TB compared to the HIV-1/non-TB. For HIV-1/TB, the average rate of evolution of C2V5 env was higher than previous reports (2.4 × 10(-4) substitution/site/day). Two groups of HIV-1/TB were observed based on the rate of HIV-1 evolution and coreceptor usage: A fast evolving R5-tropic dominating group and a relatively slowly evolving X4 group. The results demonstrated that active TB has an impact on HIV-1 viral diversity and divergence over time. The influence of active TB on longitudinal evolution of HIV-1 may be predominant for R5 viruses.
Asunto(s)
Evolución Molecular , Infecciones por VIH/complicaciones , Infecciones por VIH/virología , VIH-1/clasificación , VIH-1/genética , Tuberculosis/complicaciones , Adulto , Antituberculosos/uso terapéutico , Clonación Molecular , Femenino , Genotipo , VIH-1/aislamiento & purificación , Humanos , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Plasma/virología , Reacción en Cadena de la Polimerasa , Polimorfismo Genético , ARN Viral/genética , ARN Viral/aislamiento & purificación , Receptores del VIH , Análisis de Secuencia de ADN , Tuberculosis/tratamiento farmacológico , Acoplamiento Viral , Productos del Gen env del Virus de la Inmunodeficiencia Humana/genéticaAsunto(s)
Síndrome de Inmunodeficiencia Adquirida/inmunología , Fármacos Anti-VIH/administración & dosificación , Recuento de Linfocito CD4 , Inhibidores de la Proteasa del VIH/administración & dosificación , VIH-1 , Síndrome de Inmunodeficiencia Adquirida/tratamiento farmacológico , Síndrome de Inmunodeficiencia Adquirida/mortalidad , Quimioterapia Combinada , Humanos , Factores de RiesgoRESUMEN
Human immunodeficiency virus (HIV)-infected, active intravenous drug users received once-daily therapy consisting of a combination of didanosine (2',3'-dideoxyinosine or DDI), lamivudine [(-)-beta-L-2',3'-dideoxy-3'-thiacytidine or 3TC] and nevirapine. Preliminary results for the first 24 weeks show that the regimen provides potent immunological and antiviral effects.