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Coxsackievirus-adenovirus receptor genetically fused to anti-human CD40 scFv enhances adenoviral transduction of dendritic cells

Gene Therapy volume 9, pages 1189–1193 (2002)Cite this article

Abstract

A promising approach to immunotherapy involves the loading of dendritic cells (DCs) with genetic material to facilitate sustained expression of a relevant antigen in this population of potent antigen presenting cells (APC). Viral vectors such as adenovirus (Ad) have been used for this purpose. Existing methods for DC infection are limited by lack of specificity and a requirement for DC exposure to high viral doses. Targeting of Ad to DCs with bispecific antibodies has significantly augmented levels of transgene expression. Genetic fusion of the extracellular portion of coxsackievirus-adenovirus receptor (CAR) to cell-specific ligands has also proved successful in targeting Ad to cells of interest. We report here the production and primary characterization of a new fusion protein comprising the ecto-domain of CAR connected to a single chain antibody (scFv) G28-5 against human CD40 present on the surface of DCs. We demonstrate that the fusion protein (CAR/G28) specifically interacts with both recombinant Ad fiber knob and the ecto-domain of human CD40 in a binding assay (ELISA). Finally, we show that the CAR/G28 fusion protein promotes highly efficient transduction of DCs of both rhesus monkey and human origin.

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Acknowledgements

This work was sponsored by the grant DAMD17-00-1-0115 from United States Army Department of Defense and the grants R01 CA86881, R21 AI44322, U19 DK57858, N01 CO-97110 R01 HL67962 from the National Institute of Health. The authors would like to thank Drs Paul Reynolds and Laura Timares for fruitful discussions.

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Authors and Affiliations

  1. Division of Human Gene Therapy, Departments of Medicine, Pathology and Surgery, University of Alabama at Birmingham, Birmingham, AL, USA

    A V Pereboev, Y Kawakami, J L Blackwell, E A Kashentseva, D T Curiel & I P Dmitriev

  2. Gene Therapy Center, University of Alabama at Birmingham, Birmingham, AL, USA

    A V Pereboev, Y Kawakami, J L Blackwell, E A Kashentseva, P L Triozzi, D T Curiel, J M Thomas & I P Dmitriev

  3. Division of Hematology-Oncology, University of Alabama at Birmingham, Birmingham, AL, USA

    P L Triozzi & W A Aldrich

  4. Division of Transplantation, University of Alabama at Birmingham, Birmingham, AL, USA

    C K Asiedu, S S Dong & J M Thomas

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  1. A V Pereboev
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Pereboev, A., Asiedu, C., Kawakami, Y. et al. Coxsackievirus-adenovirus receptor genetically fused to anti-human CD40 scFv enhances adenoviral transduction of dendritic cells. Gene Ther 9, 1189–1193 (2002). https://doi.org/10.1038/sj.gt.3301767

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