Limits...
Anti-gp120 minibody gene transfer to female genital epithelial cells protects against HIV-1 virus challenge in vitro.

Abdel-Motal UM, Sarkis PT, Han T, Pudney J, Anderson DJ, Zhu Q, Marasco WA - PLoS ONE (2011)

Bottom Line: Accordingly, a recombinant AAV vector that encodes human b12 anti-HIV gp120 BnAb as a single-chain variable fragment Fc fusion (scFvFc), or "minibody" was constructed.Furthermore, cervico-vaginal epithelial stem cells were found to be efficiently transduced by the optimal AAV serotype mediated expression of GFP.This study provides the foundation for a novel microbicide strategy to protect against sexual transmission of HIV-1 by AAV transfer of broadly neutralizing antibody genes to cervico-vaginal epithelial stem cells that could replenish b12 BnAb secreting cells through multiple menstrual cycles.

View Article: PubMed Central - PubMed

Affiliation: Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts, United States of America.

ABSTRACT

Background: Although cervico-vaginal epithelial cells of the female lower genital tract provide the initial defense system against HIV-1 infection, the protection is sometimes incomplete. Thus, enhancing anti-HIV-1 humoral immunity at the mucosal cell surface by local expression of anti-HIV-1 broadly neutralizing antibodies (BnAb) that block HIV-1 entry would provide an important new intervention that could slow the spread of HIV/AIDS.

Methods and findings: This study tested the hypothesis that adeno-associated virus (AAV)-BnAb gene transfer to cervico-vaginal epithelial cells will lead to protection against HIV-1. Accordingly, a recombinant AAV vector that encodes human b12 anti-HIV gp120 BnAb as a single-chain variable fragment Fc fusion (scFvFc), or "minibody" was constructed. The secreted b12 minibody was shown to be biologically functional in binding to virus envelope protein, neutralizing HIV-1 and importantly, blocking transfer and infectivity of HIV-1(bal) in an organotypic human vaginal epithelial cell (VEC) model. Furthermore, cervico-vaginal epithelial stem cells were found to be efficiently transduced by the optimal AAV serotype mediated expression of GFP.

Conclusion: This study provides the foundation for a novel microbicide strategy to protect against sexual transmission of HIV-1 by AAV transfer of broadly neutralizing antibody genes to cervico-vaginal epithelial stem cells that could replenish b12 BnAb secreting cells through multiple menstrual cycles.

Show MeSH

Related in: MedlinePlus

AAV-6-GFP transduction of human primary genital epithelial cells was assessed by flow cytometry to detect GFP expression.(A) untreated cells. (B) transduced with AAV-6-GFP.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3198777&req=5

pone-0026473-g002: AAV-6-GFP transduction of human primary genital epithelial cells was assessed by flow cytometry to detect GFP expression.(A) untreated cells. (B) transduced with AAV-6-GFP.

Mentions: Based on the experiments above (Fig. 1A, B), AAV-6-GFP was chosen as the most efficient serotype to test transduction of huPGEC. Single cell suspensions of huPGEC were prepared from discarded cervical tissues from anonymous hysterectomy patients and cultured in serum-free keratinocyte medium as described in Materials and Methods. Only low passage (2–3 passages) huPGEC were used for transduction. Cells were exposed to AAV-6-GFP and found to be efficiently transduced as determined from flow cytometric analysis of total primary cervico-vaginal cells (Fig. 2A-B) and visual assessment by fluorescence microscopy (data not shown).


Anti-gp120 minibody gene transfer to female genital epithelial cells protects against HIV-1 virus challenge in vitro.

Abdel-Motal UM, Sarkis PT, Han T, Pudney J, Anderson DJ, Zhu Q, Marasco WA - PLoS ONE (2011)

AAV-6-GFP transduction of human primary genital epithelial cells was assessed by flow cytometry to detect GFP expression.(A) untreated cells. (B) transduced with AAV-6-GFP.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3198777&req=5

pone-0026473-g002: AAV-6-GFP transduction of human primary genital epithelial cells was assessed by flow cytometry to detect GFP expression.(A) untreated cells. (B) transduced with AAV-6-GFP.
Mentions: Based on the experiments above (Fig. 1A, B), AAV-6-GFP was chosen as the most efficient serotype to test transduction of huPGEC. Single cell suspensions of huPGEC were prepared from discarded cervical tissues from anonymous hysterectomy patients and cultured in serum-free keratinocyte medium as described in Materials and Methods. Only low passage (2–3 passages) huPGEC were used for transduction. Cells were exposed to AAV-6-GFP and found to be efficiently transduced as determined from flow cytometric analysis of total primary cervico-vaginal cells (Fig. 2A-B) and visual assessment by fluorescence microscopy (data not shown).

Bottom Line: Accordingly, a recombinant AAV vector that encodes human b12 anti-HIV gp120 BnAb as a single-chain variable fragment Fc fusion (scFvFc), or "minibody" was constructed.Furthermore, cervico-vaginal epithelial stem cells were found to be efficiently transduced by the optimal AAV serotype mediated expression of GFP.This study provides the foundation for a novel microbicide strategy to protect against sexual transmission of HIV-1 by AAV transfer of broadly neutralizing antibody genes to cervico-vaginal epithelial stem cells that could replenish b12 BnAb secreting cells through multiple menstrual cycles.

View Article: PubMed Central - PubMed

Affiliation: Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts, United States of America.

ABSTRACT

Background: Although cervico-vaginal epithelial cells of the female lower genital tract provide the initial defense system against HIV-1 infection, the protection is sometimes incomplete. Thus, enhancing anti-HIV-1 humoral immunity at the mucosal cell surface by local expression of anti-HIV-1 broadly neutralizing antibodies (BnAb) that block HIV-1 entry would provide an important new intervention that could slow the spread of HIV/AIDS.

Methods and findings: This study tested the hypothesis that adeno-associated virus (AAV)-BnAb gene transfer to cervico-vaginal epithelial cells will lead to protection against HIV-1. Accordingly, a recombinant AAV vector that encodes human b12 anti-HIV gp120 BnAb as a single-chain variable fragment Fc fusion (scFvFc), or "minibody" was constructed. The secreted b12 minibody was shown to be biologically functional in binding to virus envelope protein, neutralizing HIV-1 and importantly, blocking transfer and infectivity of HIV-1(bal) in an organotypic human vaginal epithelial cell (VEC) model. Furthermore, cervico-vaginal epithelial stem cells were found to be efficiently transduced by the optimal AAV serotype mediated expression of GFP.

Conclusion: This study provides the foundation for a novel microbicide strategy to protect against sexual transmission of HIV-1 by AAV transfer of broadly neutralizing antibody genes to cervico-vaginal epithelial stem cells that could replenish b12 BnAb secreting cells through multiple menstrual cycles.

Show MeSH
Related in: MedlinePlus