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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.

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Related in: MedlinePlus

Adenoviral E1A copy numbers in transduced human CD8+ T cells.Increased adenoviral E1A gene copy number in Ad-RGD treated cells (black bar) in comparison to Ad-WT treated cells (white bar) at different time-points after virus transduction calculated by qPCR. Samples 1–3 are three independent experiments. (*p<0.001; ** p<0.05). Error bars represent mean ± SD.
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pone-0018091-g004: Adenoviral E1A copy numbers in transduced human CD8+ T cells.Increased adenoviral E1A gene copy number in Ad-RGD treated cells (black bar) in comparison to Ad-WT treated cells (white bar) at different time-points after virus transduction calculated by qPCR. Samples 1–3 are three independent experiments. (*p<0.001; ** p<0.05). Error bars represent mean ± SD.

Mentions: Similar results were observed when primary human CD8+ T cells were treated with Ad-WT and Ad-RGD. E1A copy numbers of Ad-RGD virus were significantly higher than Ad-WT as early as 24 h post-transduction and increased even more by 48 h (Fig. 4).


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)

Adenoviral E1A copy numbers in transduced human CD8+ T cells.Increased adenoviral E1A gene copy number in Ad-RGD treated cells (black bar) in comparison to Ad-WT treated cells (white bar) at different time-points after virus transduction calculated by qPCR. Samples 1–3 are three independent experiments. (*p<0.001; ** p<0.05). Error bars represent mean ± SD.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0018091-g004: Adenoviral E1A copy numbers in transduced human CD8+ T cells.Increased adenoviral E1A gene copy number in Ad-RGD treated cells (black bar) in comparison to Ad-WT treated cells (white bar) at different time-points after virus transduction calculated by qPCR. Samples 1–3 are three independent experiments. (*p<0.001; ** p<0.05). Error bars represent mean ± SD.
Mentions: Similar results were observed when primary human CD8+ T cells were treated with Ad-WT and Ad-RGD. E1A copy numbers of Ad-RGD virus were significantly higher than Ad-WT as early as 24 h post-transduction and increased even more by 48 h (Fig. 4).

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