Limits...
Enhanced transduction and replication of RGD-fiber modified adenovirus in primary T cells.

Sengupta S, Ulasov IV, Thaci B, Ahmed AU, Lesniak MS - PLoS ONE (2011)

Bottom Line: Infection with replication-competent Ad-RGD virus also caused increased cell cycling, higher E1A copy number and enriched hexon antigen expression in both human and murine T cells.In vivo, 35-45% of splenic T cells were transduced by Ad-RGD.Collectively, our results prove that a fiber modified Ad-RGD successfully transduces and replicates in primary T cells of both murine and human origin.

View Article: PubMed Central - PubMed

Affiliation: The Brain Tumor Center, The University of Chicago, Chicago, Illinois, United States of America.

ABSTRACT

Background: Adenoviruses are often used as vehicles to mediate gene delivery for therapeutic purposes, but their research scope in hematological cells remains limited due to a narrow choice of host cells that express the adenoviral receptor (CAR). T cells, which are attractive targets for gene therapy of numerous diseases, remain resistant to adenoviral infection because of the absence of CAR expression. Here, we demonstrate that this resistance can be overcome when murine or human T cells are transduced with an adenovirus incorporating the RGD-fiber modification (Ad-RGD).

Methodology/principal finding: A luciferase-expressing replication-deficient Ad-RGD infected 3-fold higher number of activated primary T cells than an adenovirus lacking the RGD-fiber modification in vitro. Infection with replication-competent Ad-RGD virus also caused increased cell cycling, higher E1A copy number and enriched hexon antigen expression in both human and murine T cells. Transduction with oncolytic Ad-RGD also resulted in higher titers of progeny virus and enhanced the killing of T cells. In vivo, 35-45% of splenic T cells were transduced by Ad-RGD.

Conclusions: Collectively, our results prove that a fiber modified Ad-RGD successfully transduces and replicates in primary T cells of both murine and human origin.

Show MeSH

Related in: MedlinePlus

β3 and β5 integrin expression on primary murine T cells.(A) Flow cytometry profile of β3 and β5 integrin expression on naïve and activated primary murine T cells. Left panel shows histograms for β3 integrins on CD4+ (top) and CD8+ T cells (bottom). Right panel shows β5 integrins on CD4+ and CD8+ T cells. (B) Bar diagram representation of β3 and β5 integrin expression on T cells. Top panel shows integrin expression on CD4+ T cells while bottom panel represents CD8+ T cells. White bars indicate naïve T cells and black bars are activated T cells. Error bars indicate mean ± SD. (*p<0.05)
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3065494&req=5

pone-0018091-g001: β3 and β5 integrin expression on primary murine T cells.(A) Flow cytometry profile of β3 and β5 integrin expression on naïve and activated primary murine T cells. Left panel shows histograms for β3 integrins on CD4+ (top) and CD8+ T cells (bottom). Right panel shows β5 integrins on CD4+ and CD8+ T cells. (B) Bar diagram representation of β3 and β5 integrin expression on T cells. Top panel shows integrin expression on CD4+ T cells while bottom panel represents CD8+ T cells. White bars indicate naïve T cells and black bars are activated T cells. Error bars indicate mean ± SD. (*p<0.05)

Mentions: Naïve and activated murine T cells were tested for the expression of β3 and β5 integrins on their surface. These integrins are components of cell-surface receptors for Ad-RGD viruses. Flow cytometric analysis showed 3.5-fold increase in mean fluorescence intensity of β3 and 1.5-fold increase in β5 expression on activated CD4+ T cells over naïve cells. In CD8+ T cells, 2.5-fold increase in both β3 and β5 expression was observed after activation with anti-CD3 antibody (Fig. 1).


Enhanced transduction and replication of RGD-fiber modified adenovirus in primary T cells.

Sengupta S, Ulasov IV, Thaci B, Ahmed AU, Lesniak MS - PLoS ONE (2011)

β3 and β5 integrin expression on primary murine T cells.(A) Flow cytometry profile of β3 and β5 integrin expression on naïve and activated primary murine T cells. Left panel shows histograms for β3 integrins on CD4+ (top) and CD8+ T cells (bottom). Right panel shows β5 integrins on CD4+ and CD8+ T cells. (B) Bar diagram representation of β3 and β5 integrin expression on T cells. Top panel shows integrin expression on CD4+ T cells while bottom panel represents CD8+ T cells. White bars indicate naïve T cells and black bars are activated T cells. Error bars indicate mean ± SD. (*p<0.05)
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3065494&req=5

pone-0018091-g001: β3 and β5 integrin expression on primary murine T cells.(A) Flow cytometry profile of β3 and β5 integrin expression on naïve and activated primary murine T cells. Left panel shows histograms for β3 integrins on CD4+ (top) and CD8+ T cells (bottom). Right panel shows β5 integrins on CD4+ and CD8+ T cells. (B) Bar diagram representation of β3 and β5 integrin expression on T cells. Top panel shows integrin expression on CD4+ T cells while bottom panel represents CD8+ T cells. White bars indicate naïve T cells and black bars are activated T cells. Error bars indicate mean ± SD. (*p<0.05)
Mentions: Naïve and activated murine T cells were tested for the expression of β3 and β5 integrins on their surface. These integrins are components of cell-surface receptors for Ad-RGD viruses. Flow cytometric analysis showed 3.5-fold increase in mean fluorescence intensity of β3 and 1.5-fold increase in β5 expression on activated CD4+ T cells over naïve cells. In CD8+ T cells, 2.5-fold increase in both β3 and β5 expression was observed after activation with anti-CD3 antibody (Fig. 1).

Bottom Line: Infection with replication-competent Ad-RGD virus also caused increased cell cycling, higher E1A copy number and enriched hexon antigen expression in both human and murine T cells.In vivo, 35-45% of splenic T cells were transduced by Ad-RGD.Collectively, our results prove that a fiber modified Ad-RGD successfully transduces and replicates in primary T cells of both murine and human origin.

View Article: PubMed Central - PubMed

Affiliation: The Brain Tumor Center, The University of Chicago, Chicago, Illinois, United States of America.

ABSTRACT

Background: Adenoviruses are often used as vehicles to mediate gene delivery for therapeutic purposes, but their research scope in hematological cells remains limited due to a narrow choice of host cells that express the adenoviral receptor (CAR). T cells, which are attractive targets for gene therapy of numerous diseases, remain resistant to adenoviral infection because of the absence of CAR expression. Here, we demonstrate that this resistance can be overcome when murine or human T cells are transduced with an adenovirus incorporating the RGD-fiber modification (Ad-RGD).

Methodology/principal finding: A luciferase-expressing replication-deficient Ad-RGD infected 3-fold higher number of activated primary T cells than an adenovirus lacking the RGD-fiber modification in vitro. Infection with replication-competent Ad-RGD virus also caused increased cell cycling, higher E1A copy number and enriched hexon antigen expression in both human and murine T cells. Transduction with oncolytic Ad-RGD also resulted in higher titers of progeny virus and enhanced the killing of T cells. In vivo, 35-45% of splenic T cells were transduced by Ad-RGD.

Conclusions: Collectively, our results prove that a fiber modified Ad-RGD successfully transduces and replicates in primary T cells of both murine and human origin.

Show MeSH
Related in: MedlinePlus