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Adenovirus-mediated expresssion of the murine ecotropic receptor facilitates transduction of human hematopoietic cells with an ecotropic retroviral vector

Gene Therapy volume 6, pages 1456–1468 (1999)Cite this article

Abstract

One factor limiting the ability to modify human repopulating hematopoietic cells genetically with retroviral vectors is the relatively low expression of the cognate viral receptor. We have tested sequential transduction of human hematopoietic cells with an adenoviral vector encoding the ecotropic retroviral receptor followed by transduction with an ecotropic retroviral vector. Adenoviral transduction of K562 erythroleukemia cells was highly efficiently with >95% of cells expressing the ecotropic receptor at a multiplicity of infection (MOI) of 103with a correspondingly high transduction with a retroviral vector. Ecotropic receptor expression in CD34+ cells following transduction with adenoviral vectors was increased by at least two-fold (from 20 to 48%) by replacing the RSV promoter with the CMV E1a promoter, resulting in a parallel increase in retroviral transduction efficiency. Replacing the head portion of the fiber protein in conventional adenoviral vectors (serotype 5) with the corresponding portion from an adenoviral 3 serotype resulted in ecotropic receptor expression in 60% of CD34+ cells at an MOI of 104 and a retroviral transduction of 60% of hematopoietic clonogenic progenitors. The sequential transduction strategy also resulted in efficient transduction of the primitive CD34+CD38− subset suggesting that it may hold promise for genetic modification of human hematopoietic stem cells.

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Acknowledgements

We wish to thank Jean Johnson for her outstanding assistance in preparation of the manuscript, Craig Jordan for the gift of an Ad5 CMV-GFP vector, and Dr Richard Ashman for his help with the FACs analysis and figures. This work was supported by NHLBI Program Project Grant P01 HL 53749, the ASSISI Foundation of Memphis Grant 94–00, Cancer Center Support CORE Grant, P30 CA 21765 and American Lebanese Syrian Associated Charities (ALSAC). Amit C Nathwani was supported by a grant from the Wellcome Trust; number 049894/114.

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Authors and Affiliations

  1. Division of Experimental Hematology, Department of Hematology/Oncology, St Jude Children’s Research Hospital, Memphis, TN, USA

    A C Nathwani, D A Persons, P Frare, A W Nienhuis & E F Vanin

  2. Genetic Therapy Inc/a Novartis Company, Gaithersburg, MD, USA

    S C Stevenson & A McClelland

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  1. A C Nathwani
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  2. D A Persons
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  3. S C Stevenson
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  6. A W Nienhuis
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  7. E F Vanin
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Nathwani, A., Persons, D., Stevenson, S. et al. Adenovirus-mediated expresssion of the murine ecotropic receptor facilitates transduction of human hematopoietic cells with an ecotropic retroviral vector. Gene Ther 6, 1456–1468 (1999). https://doi.org/10.1038/sj.gt.3300974

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