ABSTRACT
In this work, a total of 200 samples from the HIV-infected individuals were analyzed: 50 samples from the Saha Republic (Yakutia), 50 samples from the Vologda Region (City of Cherepovets), and 100 samples from the Moscow Region (Moscow and Moscow Region). All samples were obtained from the patients who were not undergoing antiretroviral therapy. It was detected that the regulatory genes vif, vpr, vpu, rev, tat, and nef were amplified with moderate sensitivity after one-stage amplification. When those samples were analyzed by the nested PCR the detection ratio was much higher. While studying nef-gene the phenomena of the splicing in cells cores was detected at the advanced stages of the HIV-infection (3 and 4 stages). At the same time, the splicing was not detected at the earlier stages of the HIV-infection. This effect might be the cause of the transition from asymptomatic stage of the infection to the advanced stage. It was also shown for the first time that the variability of the regulatory genes correlated with the virus subtype.
Subject(s)
Gene Expression Regulation, Viral , Genes, Regulator , Genes, Viral , HIV Infections/pathology , HIV-1/genetics , DNA Primers , Disease Progression , Female , Genotype , HIV Infections/virology , Humans , Male , Molecular Typing , Moscow , Polymerase Chain Reaction , RNA Splicing , Russia , SiberiaABSTRACT
A panel of 16 HIV-1 isolates was designed. Those isolates were isolated from patients undergoing HAART and developing resistance to the antiretroviral drugs. It was shown that the isolates were resistant to nucleoside RT inhibitors (retrovir, epivir) and non-nucleoside inhibitors (viramun). Isolates had stable replication activity. Average rate of cells expressing viral Ag was 14-20%. The infectious titer was 2.4 Ig TCID50. The sequencing showed that all isolates were of the subtype A dominating in the major part of Russian Federation. This panel could be used as the biotechnological base for studying antiretroviral drugs of new generation and for the design of experimental vaccines.
Subject(s)
Biological Specimen Banks , Drug Resistance, Viral , HIV-1/isolation & purification , Reverse Transcriptase Inhibitors , Adult , Antiretroviral Therapy, Highly Active , Female , HIV Infections/drug therapy , HIV Infections/epidemiology , Humans , Male , Russia/epidemiologyABSTRACT
Biological properties of HIV-1 laboratory strains and isolates were studies and compared with research results of the nef gene fragment obtained from their proviral DNA and RNA as well as from RNA and HIV-1 DNA isolated immediately from the blood of patients at early HIV stages. The electrophoresis pattern as well as the results of determination of nucleotide sequences showed that the HVI-1 RNA nef gene of rapid/high laboratory strains of HIV isolates and RNA obtained from patients at late infection stages had a 135-nucleotide inner deletion. The phenomenon can be regarded is proof of that nef gene structure, if violated, causes an enhanced virulence of and an intensified multiplication of the virus (according to laboratory markers) observed at late HIV stages, which triggers, at least in a number of cases, the infection aggravation.
