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Patients infected with CRF07_BC have significantly lower viral loads than patients with HIV-1 subtype B: mechanism and impact on disease progression.

Huang SW, Wang SF, Lin YT, Yen CH, Lee CH, Wong WW, Tsai HC, Yang CJ, Hu BS, Lin YH, Wang CT, Wang JJ, Hu Z, Kuritzkes DR, Chen YH, Chen YM - PLoS ONE (2014)

Bottom Line: The replicative capacity of nine CRF07_BC and four subtype B isolates were compared and the results showed that the former had significantly lower replicative capacity than the latter although all of them were CCR5- tropic and non-syncytium inducing viruses.The results showed that 7d virus had significantly lower replication capacity, poorer protease-mediated processing and viral proteins production.In conclusion, patients infected with CRF07_BC had significantly lower viral loads than patients infected with subtype B and it may due to the deletion of 7 amino acids which overlaps with Alix protein-binding domain of the p6gag.

View Article: PubMed Central - PubMed

Affiliation: Center for Infectious Disease and Cancer Research (CICAR), Kaohsiung Medical University, Kaohsiung, Taiwan; Institute of Microbiology and Immunology, National Yang-Ming University, Taipei, Taiwan.

ABSTRACT
The circulating recombinant form (CRF) 07_BC is the most prevalent HIV-1 strain among injection drug users (IDUs) in Taiwan. It contains a 7 amino-acid deletion in its p6gag. We conducted a cohort study to compare viral loads and CD4 cell count changes between patients infected with subtype B and CRF07_BC and to elucidate its mechanism. Twenty-one patients infected with CRF07_BC and 59 patients with subtype B were selected from a cohort of 667 HIV-1/AIDS patients whom have been followed up for 3 years. Generalized estimated equation was used to analyze their clinical data and the results showed that patients infected with CRF07_BC had significantly lower viral loads (about 58,000 copies per ml less) than patients with subtype B infection (pā€Š=ā€Š0.002). The replicative capacity of nine CRF07_BC and four subtype B isolates were compared and the results showed that the former had significantly lower replicative capacity than the latter although all of them were CCR5- tropic and non-syncytium inducing viruses. An HIV-1-NL4-3 mutant virus which contains a 7 amino-acid deletion in p6gag (designated as 7d virus) was generated and its live cycle was investigated. The results showed that 7d virus had significantly lower replication capacity, poorer protease-mediated processing and viral proteins production. Electron microscopic examination of cells infected with wild-type or 7d virus demonstrated that the 7d virus had poorer and slower viral maturation processes: more viruses attached to the cell membrane and higher proportion of immature virions outside the cells. The interaction between p6gag and Alix protein was less efficient in cells infected with 7d virus. In conclusion, patients infected with CRF07_BC had significantly lower viral loads than patients infected with subtype B and it may due to the deletion of 7 amino acids which overlaps with Alix protein-binding domain of the p6gag.

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The interaction between Alix protein and wild type/mutant Gag. MAGIC-5 cells were infected with wt or 7d recombinant virus for 48 hours.The Alix and Gag proteins were analyze by TIRF-SR with rabbit anti-Alix polyclonal antibody and mouse anti-p24 monoclonal antibody. Red spots indicate Alix protein. Green spots indicate either wild type or 7d Gag protein. The proportion of co-localization of Alix and Gag protein was quantified using Volocity 3D Image Analysis Software.
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pone-0114441-g004: The interaction between Alix protein and wild type/mutant Gag. MAGIC-5 cells were infected with wt or 7d recombinant virus for 48 hours.The Alix and Gag proteins were analyze by TIRF-SR with rabbit anti-Alix polyclonal antibody and mouse anti-p24 monoclonal antibody. Red spots indicate Alix protein. Green spots indicate either wild type or 7d Gag protein. The proportion of co-localization of Alix and Gag protein was quantified using Volocity 3D Image Analysis Software.

Mentions: Since the 7 amino-acid deletion overlaps with the Alix binding domain of p6gag, we conducted IFA staining with anti- p24Gag and anti-Alix antibodies and analyzed their interaction using confocal microscopic exam with super-resolution program. The results showed that the co-localization coeffcicent of Gag and Alix was significantly lower in 7d virus than in wt virus (36.4% versus 48.88%, p<0.05) (Fig. 4).


Patients infected with CRF07_BC have significantly lower viral loads than patients with HIV-1 subtype B: mechanism and impact on disease progression.

Huang SW, Wang SF, Lin YT, Yen CH, Lee CH, Wong WW, Tsai HC, Yang CJ, Hu BS, Lin YH, Wang CT, Wang JJ, Hu Z, Kuritzkes DR, Chen YH, Chen YM - PLoS ONE (2014)

The interaction between Alix protein and wild type/mutant Gag. MAGIC-5 cells were infected with wt or 7d recombinant virus for 48 hours.The Alix and Gag proteins were analyze by TIRF-SR with rabbit anti-Alix polyclonal antibody and mouse anti-p24 monoclonal antibody. Red spots indicate Alix protein. Green spots indicate either wild type or 7d Gag protein. The proportion of co-localization of Alix and Gag protein was quantified using Volocity 3D Image Analysis Software.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263662&req=5

pone-0114441-g004: The interaction between Alix protein and wild type/mutant Gag. MAGIC-5 cells were infected with wt or 7d recombinant virus for 48 hours.The Alix and Gag proteins were analyze by TIRF-SR with rabbit anti-Alix polyclonal antibody and mouse anti-p24 monoclonal antibody. Red spots indicate Alix protein. Green spots indicate either wild type or 7d Gag protein. The proportion of co-localization of Alix and Gag protein was quantified using Volocity 3D Image Analysis Software.
Mentions: Since the 7 amino-acid deletion overlaps with the Alix binding domain of p6gag, we conducted IFA staining with anti- p24Gag and anti-Alix antibodies and analyzed their interaction using confocal microscopic exam with super-resolution program. The results showed that the co-localization coeffcicent of Gag and Alix was significantly lower in 7d virus than in wt virus (36.4% versus 48.88%, p<0.05) (Fig. 4).

Bottom Line: The replicative capacity of nine CRF07_BC and four subtype B isolates were compared and the results showed that the former had significantly lower replicative capacity than the latter although all of them were CCR5- tropic and non-syncytium inducing viruses.The results showed that 7d virus had significantly lower replication capacity, poorer protease-mediated processing and viral proteins production.In conclusion, patients infected with CRF07_BC had significantly lower viral loads than patients infected with subtype B and it may due to the deletion of 7 amino acids which overlaps with Alix protein-binding domain of the p6gag.

View Article: PubMed Central - PubMed

Affiliation: Center for Infectious Disease and Cancer Research (CICAR), Kaohsiung Medical University, Kaohsiung, Taiwan; Institute of Microbiology and Immunology, National Yang-Ming University, Taipei, Taiwan.

ABSTRACT
The circulating recombinant form (CRF) 07_BC is the most prevalent HIV-1 strain among injection drug users (IDUs) in Taiwan. It contains a 7 amino-acid deletion in its p6gag. We conducted a cohort study to compare viral loads and CD4 cell count changes between patients infected with subtype B and CRF07_BC and to elucidate its mechanism. Twenty-one patients infected with CRF07_BC and 59 patients with subtype B were selected from a cohort of 667 HIV-1/AIDS patients whom have been followed up for 3 years. Generalized estimated equation was used to analyze their clinical data and the results showed that patients infected with CRF07_BC had significantly lower viral loads (about 58,000 copies per ml less) than patients with subtype B infection (pā€Š=ā€Š0.002). The replicative capacity of nine CRF07_BC and four subtype B isolates were compared and the results showed that the former had significantly lower replicative capacity than the latter although all of them were CCR5- tropic and non-syncytium inducing viruses. An HIV-1-NL4-3 mutant virus which contains a 7 amino-acid deletion in p6gag (designated as 7d virus) was generated and its live cycle was investigated. The results showed that 7d virus had significantly lower replication capacity, poorer protease-mediated processing and viral proteins production. Electron microscopic examination of cells infected with wild-type or 7d virus demonstrated that the 7d virus had poorer and slower viral maturation processes: more viruses attached to the cell membrane and higher proportion of immature virions outside the cells. The interaction between p6gag and Alix protein was less efficient in cells infected with 7d virus. In conclusion, patients infected with CRF07_BC had significantly lower viral loads than patients infected with subtype B and it may due to the deletion of 7 amino acids which overlaps with Alix protein-binding domain of the p6gag.

Show MeSH
Related in: MedlinePlus