title: HIV-1 Tropism Determines Different Mutation Profiles in Proviral DNA.
abstract: In order to establish new infections HIV-1 particles need to attach to receptors expressed on the cellular surface. HIV-1 particles interact with a cell membrane receptor known as CD4 and subsequently with another cell membrane molecule known as a co-receptor. Two major different co-receptors have been identified: C-C chemokine Receptor type 5 (CCR5) and C-X-C chemokine Receptor type 4 (CXCR4) Previous reports have demonstrated cellular modifications upon HIV-1 binding to its co-receptors including gene expression modulations. Here we investigated the effect of viral binding to either CCR5 or CXCR4 co-receptors on viral diversity after a single round of reverse transcription. CCR5 and CXCR4 pseudotyped viruses were used to infect non-stimulated and stimulated PBMCs and purified CD4 positive cells. We adopted the SOLiD methodology to sequence virtually the entire proviral DNA from all experimental infections. Infections with CCR5 and CXCR4 pseudotyped virus resulted in different patterns of genetic diversification. CCR5 virus infections produced extensive proviral diversity while in CXCR4 infections a more localized substitution process was observed. In addition, we present pioneering results of a recently developed method for the analysis of SOLiD generated sequencing data applicable to the study of viral quasi-species. Our findings demonstrate the feasibility of viral quasi-species evaluation by NGS methodologies. We presented for the first time strong evidence for a host cell driving mechanism acting on the HIV-1 genetic variability under the control of co-receptor stimulation. Additional investigations are needed to further clarify this question, which is relevant to viral diversification process and consequent disease progression.
authors.affiliation: Departamento de Microbiologia e Imunologia Veterinária, Universidade Federal Rural do Rio de Janeiro (UFRRJ), Rio de Janeiro, Brazil; Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina (EPM), Universidade Federal de São Paulo (UNIFESP), São Paulo, Brazil.
authors.forename: Sieberth
authors.initials: S
authors.lastname: Nascimento-Brito
authors.affiliation: Departamento de Medicina, EPM, UNIFESP, São Paulo, Brazil.
authors.forename: Jean
authors.initials: J
authors.lastname: Paulo Zukurov
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina (EPM), Universidade Federal de São Paulo (UNIFESP), São Paulo, Brazil.
authors.forename: Juliana T
authors.initials: JT
authors.lastname: Maricato
authors.affiliation: Genomics and Computational Biology Group, Research Center René Rachou (CPqRR), Fundação Oswaldo Cruz (FIOCRUZ), Belo Horizonte, Brazil.
authors.forename: Angela C
authors.initials: AC
authors.lastname: Volpini
authors.affiliation: Genomics and Computational Biology Group, Research Center René Rachou (CPqRR), Fundação Oswaldo Cruz (FIOCRUZ), Belo Horizonte, Brazil.
authors.forename: Anna Christina M
authors.initials: AC
authors.lastname: Salim
authors.affiliation: Genomics and Computational Biology Group, Research Center René Rachou (CPqRR), Fundação Oswaldo Cruz (FIOCRUZ), Belo Horizonte, Brazil.
authors.forename: Flávio M G
authors.initials: FM
authors.lastname: Araújo
authors.affiliation: Biosystems Informatics Group, CPqRR, FIOCRUZ, Belo Horizonte, Brazil.
authors.forename: Roney S
authors.initials: RS
authors.lastname: Coimbra
authors.affiliation: Genomics and Computational Biology Group, Research Center René Rachou (CPqRR), Fundação Oswaldo Cruz (FIOCRUZ), Belo Horizonte, Brazil.
authors.forename: Guilherme C
authors.initials: GC
authors.lastname: Oliveira
authors.affiliation: Departamento de Informática em Saúde, EPM, UNIFESP, São Paulo, Brazil; Laboratório de Biocomplexidade e Genômica Evolutiva, EPM, UNIFESP, São Paulo, Brazil.
authors.forename: Fernando
authors.initials: F
authors.lastname: Antoneli
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina (EPM), Universidade Federal de São Paulo (UNIFESP), São Paulo, Brazil; Departamento de Medicina, EPM, UNIFESP, São Paulo, Brazil.
authors.forename: Luiz Mário R
authors.initials: LM
authors.lastname: Janini
mesh_terms: D019943:Amino Acid Substitution; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D003062:Codon / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics*; D004587:Electrophoresis, Agar Gel; D005434:Flow Cytometry; D015658:HIV Infections / Q000276:immunology / Q000821:virology; D015497:HIV-1 / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D009154:Mutation / Q000235:genetics*; D009711:Nucleotides / Q000235:genetics; D016366:Open Reading Frames / Q000235:genetics; D011533:Proviruses / Q000235:genetics*; D019713:Receptors, CCR5 / Q000378:metabolism; D017422:Sequence Analysis, DNA; D013223:Statistics as Topic; D018523:Tropism / Q000235:genetics*
chemical_list: D003062:Codon; D004279:DNA, Viral; D009711:Nucleotides; D019713:Receptors, CCR5