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260 Cloning and Analysis of Full-Length Genomes of HIV-1 Strains Prevalent among IDU in Countries of the FSU
A. E. Masharsky*1, V. F. Eremin2, N. A. Klimov1, and A. P. Kozlov1
1Biomedical Ctr., St. Petersburg, Russia and 2Res. Inst. for Epidemiology and Microbiology, Minsk, Belarus
Background: The large-scale HIV-1 epidemic among IDU in countries of the FSU was caused mainly by 2 HIV-1 variants. The first variant belongs to subtype A, the second one is a recombinant between A and B subtypes and both are characterized by high homogeneity of nucleotide sequences. For detailed study of HIV-1 epidemic in countries of the FSU and for construction of HIV vaccine we have cloned and sequenced full-length genomes of HIV-1 isolates representing prevalent HIV-1 variants.
Methods: Based on V3 serotyping and V3-V5 env and p17/p24 gag sequencing, 2 representative HIV-1 isolates were selected for cloning and sequencing their full-length genomes. Amplification was carried out by nested PCR on DNA from PBMC and resulted in a set of 8 overlapping DNA fragments covering a full-length HIV-1 genome. All fragments were cloned and sequenced. Nucleotide sequences were subjected to phylogenetic and recombinant analysis and studied for biological, antigenic, and immunogenic peculiarities of HIV-1 isolates.
Results: Full-length genomes of 2 prevalent in countries of the FSU HIV-1 variants were amplified, cloned, and sequenced. Both assembled genomes contained all known active HIV-1 ORFs. Phylogenetic and recombinant analysis of full-length genome nucleotide sequences revealed that the first isolate was most similar to subtype A throughout the entire genome, whereas the second one was recombinant between A and B subtypes with a structure A/B/A. Comparative analysis showed high similarity of nucleotide sequences of selected isolates with published partial HIV sequences from countries of the FSU. We found out that the mean genetic distance between Russian and African strains of subtype A was 11.1%, which was greater than this value between African strains of subtype A (8.6%). Thus biological and antigenic properties of Russian isolates of subtype A may differ from African isolates.
Conclusions: We have cloned and sequenced full-length genomes of HIV-1 strains representing 2 prevailing Russian HIV-1 variants. They are currently used for construction of HIV vaccine.
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