Aim: Separation of HIV-1 isolates from HIV infected patients who had received antiretroviral therapy courses. Analysis of genetic and replicative properties of the separated isolates, study of pol gene mutation stability sustentation that is responsible for the emergence of drug resistance.
Materials And Methods: HIV isolate separation was carried out by co-cultivation ofperipheral blood mononuclears of HIV infected patients with previously stimulated phytohemagglutinin cells of healthy donors. Virus replication was evaluated by the level of p24 virus specific protein accumulation determined in enzyme immunoassay. HIV-1 subtype identification, detection of HIV-1 genome mutations were carried out by pol gene nucleotide sequence determination and subsequent analysis of the data obtained - by using specialized program resources.
Results: 14 infectious HIV-1 subtype A, B and CRF02_AG isolates were separated containing various sets of mutations determining resistance to widely used in clinical practice nucleoside and non-nucleoside reversetranscriptase inhibitors. Comparative analysis of mutation specter detected in HIV-1 variant genomes before isolation and after their cultivation showed that during HIV-1 cultivation in mononuclear blood cells without the addition of antiretroviral preparations not only partial loss of mutations is observed but also emergence of new drug resistance mutations; and most of the mutation causing virus resistance to antiretroviral preparations remain.
Conclusion: High reproductive properties of the HIV-1 isolates separated allow to use them to evaluate effectiveness of the drugs being developed against HIV-1 resistant to antiretroviral preparations.
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