Melo Fernando Lucas, Jamal Leda Fátima, Zanotto Paolo Marinho de Andrade
Laboratory of Molecular Evolution and Bioinformatics, Department of Microbiology, Biomedical Sciences Institute-ICBII, University of São Paulo, Brazil.
AIDS Res Hum Retroviruses. 2012 Sep;28(9):1082-8. doi: 10.1089/AID.2011.0123. Epub 2012 Apr 16.
We report for the first time the genetic and biological characterization of 10 HIV-1 primary isolates representing CRF28_BF and CRF29_BF together with additional unique BF recombinant forms (URFs) obtained by PBMC cocultivation. Recombination is an important factor promoting the increase in the genetic diversity of HIV-1. Notably, more than 20% of HIV-1 sequences worldwide were recombinants. Several recombinant viruses were reported in Brazil, and six circulating recombinant forms (CRFs) have been identified (CRF28_BF, CRF29_BF, CRF31_BC, CRF39_BF, CRF40_BF, and CRF46_BF). CRF28_BF and CRF29_BF were found to infect almost 30% of the patients in São Paulo State. The near full-length genomes of these 10 primary isolates were amplified by nested PCR in three overlapping segments, purified, and sequenced. Three samples were related to CRF28_BF, three to CRF29_BF, and four were unique recombinant forms (URFs), as determined by their breakpoint profile determined with the jpHMM program. Additionally, the coreceptor usage of these isolates was investigated in vitro using GHOST assays, which revealed three dual-tropic (X4/R5) viruses, four lymphotropic (X4) viruses, and three macrophage-tropic (R5) viruses with different V3-loop motifs, which challenges the notion that GWGR-carrying viruses are macrophage-tropic only. In sum, we report a much-anticipated well-characterized panel of viruses representing CRF28_BF, CRF29_BF, and URFs from São Paulo State, Brazil.
我们首次报告了代表CRF28_BF和CRF29_BF的10株HIV-1原代分离株以及通过PBMC共培养获得的其他独特BF重组形式（URF）的基因和生物学特征。重组是促进HIV-1基因多样性增加的一个重要因素。值得注意的是，全球超过20%的HIV-1序列是重组体。巴西报告了几种重组病毒，已鉴定出六种流行重组形式（CRF）（CRF28_BF、CRF29_BF、CRF31_BC、CRF39_BF、CRF40_BF和CRF46_BF）。发现CRF28_BF和CRF29_BF感染了圣保罗州近30%的患者。通过巢式PCR在三个重叠片段中扩增这10株原代分离株的近全长基因组，进行纯化和测序。根据用jpHMM程序确定的断点图谱，三个样本与CRF28_BF相关，三个与CRF29_BF相关，四个是独特的重组形式（URF）。此外，使用GHOST试验在体外研究了这些分离株的共受体使用情况，结果显示有三种双嗜性（X4/R5）病毒、四种嗜淋巴细胞性（X4）病毒和三种嗜巨噬细胞性（R5）病毒，它们具有不同的V3环基序，这对携带GWGR的病毒仅为嗜巨噬细胞性的观点提出了挑战。总之，我们报告了一组备受期待的、特征明确的病毒，它们代表来自巴西圣保罗州的CRF28_BF、CRF29_BF和URF。
