RESUMO
The biological characteristics of HIV-1 primary isolates of different recombinant forms (RFs) and non-B subtypes from Galicia, Spain, were investigated and the relationships between biological phenotype and evolution of infection were determined. Peripheral blood mononuclear cells (PBMCs) were obtained during the follow-up of 32 patients infected with HIV-1 non-subtype B genetic forms, characterized in partial sequences of pol (protease-reverse transcriptase) and env V3 region: 12 (37.5%) circulating RFs (CRFs), 9 (28.1%) unique RFs (URFs), and 11(34.4%) non-B subtypes. Primary isolates were obtained by coculture with donor PBMCs. Syncytium-inducing (SI) phenotype was examined in MT2 cell line and coreceptor use in GHOST and U87.CD4 cells. Fifty percent of tissue culture infective dose (TCID(50)) and viral phenotype based on V3 net charge and Geno2pheno(coreceptor) bioinformatic method were determined. Fifty-four HIV-1 primary isolates were obtained. CRF14_BG and BG URFs represented the largest group, being all SI/X4, independently of the CD4+ cell count, viral load, or the duration of infection. By contrast, 10 of 11 CRF02_AG viruses were NSI/R5. The prediction of co-receptor use was concordant with biological characterization in all NSI/R5 and in 23 of 26 SI/X4 isolates. The presence of SI/X4 or SI/X4,R5 isolates at early stages of the infection in addition to a decrease in CD4+ counts below 500 cells/microl between 2 and 6 years since diagnosis was observed in all patients infected with CRF14_BG and BG URFs. These data contrast with the usual progression in B subtype infections, in which SI/X4 viruses rarely predominate in the early years of HIV-1 infection.
Assuntos
Infecções por HIV/epidemiologia , Infecções por HIV/virologia , HIV-1/classificação , HIV-1/isolamento & purificação , Recombinação Genética , Sequência de Aminoácidos , Linhagem Celular Tumoral , Células Gigantes/fisiologia , Proteína gp120 do Envelope de HIV/genética , HIV-1/genética , HIV-1/patogenicidade , Humanos , Dados de Sequência Molecular , Fragmentos de Peptídeos/genética , Fenótipo , Análise de Sequência de DNA , Espanha/epidemiologiaRESUMO
The oligonucleotide ligation assay is a genotypic assay for the detection of resistance-associated mutations to reverse transcriptase and protease inhibitors in human immunodeficiency virus type 1 subtype B. This assay has been modified and developed for non-B subtypes and recombinant strains and has been evaluated with sequencing, resulting in a more sensitive assay than sequencing for non-B subtypes.
Assuntos
Fármacos Anti-HIV/farmacologia , DNA Ligases/metabolismo , Farmacorresistência Viral/genética , Inibidores da Protease de HIV/farmacologia , HIV-1/efeitos dos fármacos , Oligonucleotídeos/metabolismo , Mutação Puntual , Inibidores da Transcriptase Reversa/farmacologia , Produtos do Gene pol/genética , Infecções por HIV/virologia , HIV-1/classificação , HIV-1/genética , Humanos , Testes de Sensibilidade Microbiana/métodos , Oligonucleotídeos/química , Oligonucleotídeos/genética , RNA Viral/sangue , Recombinação Genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sensibilidade e Especificidade , Fatores de TempoAssuntos
Infecções por HIV/virologia , HIV-1/genética , Proteína gp120 do Envelope de HIV/genética , HIV-1/isolamento & purificação , HIV-1/metabolismo , Humanos , Fragmentos de Peptídeos/genética , Receptores de Quimiocinas/genética , Receptores de Quimiocinas/metabolismo , Receptores de HIV/genética , Receptores de HIV/metabolismo , Recombinação Genética , Espanha , Células Tumorais CultivadasRESUMO
The findings that BF intersubtype recombinant human immunodeficiency type 1 viruses (HIV-1) with coincident breakpoints in pol are circulating widely in Argentina and that non-recombinant F subtype viruses have failed to be detected in this country were reported recently. To analyse the mosaic structures of these viruses and to determine their phylogenetic relationship, near full-length proviral genomes of eight of these recombinant viruses were amplified by PCR and sequenced. Intersubtype breakpoints were analysed by bootscanning and examining the signature nucleotides. Phylogenetic relationships were determined with neighbour-joining trees. Five viruses, each with predominantly subtype F genomes, exhibited mosaic structures that were highly similar. Two intersubtype breakpoints were shared by all viruses and seven by the majority. Of the consensus breakpoints, all nine were present in two viruses, which exhibited identical recombinant structures, and four to eight breakpoints were present in the remaining viruses. Phylogenetic analysis of partial sequences supported both a common ancestry, at least in part of their genomes, for all recombinant viruses and the phylogenetic relationship of F subtype segments with F subtype viruses from Brazil. A common ancestry of the recombinants was supported also by the presence of shared signature amino acids and nucleotides, either unreported or highly unusual in F and B subtype viruses. These results indicate that HIV-1 BF recombinant viruses with diverse mosaic structures, including a circulating recombinant form (which are widespread in Argentina) derive from a common recombinant ancestor and that F subtype segments of these recombinants are related phylogenetically to the F subtype viruses from Brazil.