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  • Viral Transfer Technology
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Differences among nonhuman primates in susceptibility to bone marrow progenitor transduction with retrovirus vectors

Gene Therapy volume 7, pages 359–367 (2000)Cite this article

Abstract

Nonhuman primates are increasingly being used as models for pre-clinical assessment of retrovirus vector expression and function following stem and progenitor cell transduction. We compared the relative susceptibility of CD34+ marrow progenitors from four nonhuman primate species and humans to transduction with amphotropic pseudotyped retrovirus vectors containing the Neo gene. The rate of functional gene transfer was measured by colony formation under G418 selection. Marrow progenitors from pigtail macaques (Macaca nemestrina) were transduced at about twice the rate (19.1 ± 4.3%) as those from rhesus (11.2 ± 3.7%) and cynomolgus (7.6 ± 1.9%) macaques, baboons (7.8 ± 1.8%), and humans (9.6 ± 1.7%). Semiquantitative RT/PCR analysis suggests this difference may be due to elevated expression of the amphotropic receptor Pit2 in pigtailed macaque CD34+ cells. Further, transduction rates increased an average 1.6 ± 0.4-fold when the culture temperature was lowered to 33°C, and 2.1 ± 0.3-fold when the culture dishes were coated with the fibronectin fragment CH-296. The data presented here point to important differences among nonhuman primate models as well as transduction culture conditions, and suggest that pigtailed macaques may be particularly useful for assessing expression and function of therapeutic retrovirus vectors.

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Acknowledgements

We would like to thank A Dusty Miller for the retrovirus vector LN and packaging line PA317, Kateri A Moore for the N2/AM12 producer clone, and Kenneth Kaushansky for the gibbon IL-3 producing cell line BHKIL-3 and recombinant human Tpo. We would like to acknowledge the following companies for providing biological materials: Amgen Corp. for recombinant human SCF; Sandoz Pharmaceuticals Corp. for recombinant human IL-6; Immunex Corp. for recombinant human Flt3-L; Genetics Institute for recombinant human IL-11; and Takara Shuzo Co., Ltd, for the recombinant fibronectin fragment CH-296. We also gratefully thank XiaoChun Wang, Arron Winkler and Betty Mastropaolo for technical support. This work was made possible in part through the tissue distribution program of the University of Washington Regional Primate Center (NIH grant RR00166). This work was supported by grants and contracts HL 53750, AI35191, and HL 54881 from the National Institutes of Health.

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Authors and Affiliations

  1. Department of Medicine, University of Washington, Seattle, WA, USA

    D W Emery & T Papayannopoulou

  2. University of Washington Regional Primate Research Center, Seattle, WA, USA

    D W Emery, R G Andrews & T Papayannopoulou

  3. Department of Pediatrics, University of Washington, Seattle, WA, USA

    R G Andrews

  4. Fred Hutchinson Cancer Research Center, Seattle, WA, USA

    R G Andrews

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Emery, D., Andrews, R. & Papayannopoulou, T. Differences among nonhuman primates in susceptibility to bone marrow progenitor transduction with retrovirus vectors. Gene Ther 7, 359–367 (2000). https://doi.org/10.1038/sj.gt.3301107

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