Safe, Efficient Production of Retroviral Vectors

  • H. Kotani
  • G. J. Mcgarrity
Part of the Handbook of Experimental Pharmacology book series (HEP, volume 137)


The first clinical trials in gene therapy were held in the United States in 1990 at the National Institutes of Health (NIH). The first trial was for gene marking. Lymphocytes were removed from a melanoma biopsy, expanded in vitro and transduced with a retroviral vector carrying a gene that conferred resistance to the neomycin analogue, G-418. The clinical objective was to determine the distribution and longevity of these marked lymphocytes. Shortly thereafter, on 14 September 1990, the first clinical trial in gene therapy was initiated. A child with a form of severe combined immune deficiency (SCID), due to a defect in the adenosine deaminase (ADA) gene, was re-infused with her own lymphocytes which had been propagated ex vivo and transduced with the retroviral vector (LASN) that contained the wild-type ADA gene. Results of this and a related ADA trial, which involved the introduction of the gene into CD34+ cells from umbilical-cord blood obtained from neonates, have been published (BLAESE et al. 1995; KOHN et al. 1995).


Packaging Cell Severe Combine Immune Deficiency Producer Cell Line Roller Bottle Cell Cube 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Anderson WF, McGarrity GJ, Moen RC (1993) Report to the NIH recombinant DNA advisory committee on murine replication-competent retrovirus assays. Hum Gene Ther 4: 311 – 321PubMedCrossRefGoogle Scholar
  2. Anonymous (1997) Human gene marker/therapy clinical protocols. Hum Gene Ther 8: 2301 – 2338CrossRefGoogle Scholar
  3. Blaese RM, Culver KW, Miller AD, Carter C et al. (1995) T lymphocyte-directed gene therapy for ADA-SCID: initial trial results after 4 years. Science 270: 475 – 480PubMedCrossRefGoogle Scholar
  4. Blaese RM (1997) Gene therapy for cancer. Sci Am 276: 111 – 115PubMedCrossRefGoogle Scholar
  5. Center for Disease Control/National Instututes of Health (1993) Biological safety on microbiological and biomedical laboratories. US Health and Human Services Publication No (CDC) 93-8395. US PHS Washington, DCGoogle Scholar
  6. Chong H, Vile RG (1996) Replication-competent retrovirus produced by a “split- function” third generation amphotropic packaging cell line. Gene Ther 3: 623 – 629Google Scholar
  7. Chuck AS, Paulsson BO (1996) Consistent and high rates of gene transfer can be obtained using flow through transduction over a wide range of retroviral vectors. Hum Gene Ther 7: 743 – 750PubMedCrossRefGoogle Scholar
  8. Cornetta K, Moen RC, Culver K, Morgan RA, McLachlin JR, Sturm S, Selegue J, London W, Blaese RM, Anderson WF (1990) Amphotropic murine leukemia retrovirus is not an acute pathogen for primates. Hum Gene Ther 1: 13 – 26CrossRefGoogle Scholar
  9. Danos O, Mulligan RC (1988) Safe and efficient generation of recombinant retroviruses with amphotropic and ecotropic host ranges. Proc Natl Acad Sci USA 85: 6460 – 6464PubMedCrossRefGoogle Scholar
  10. Donahue RE, Kessler SW, Bodine D, McDonagh K, Byrne E, Raffield M, Moen R, Bacher J, Zsebo KM, Nienhuis AW (1992) Helper virus induced T cell lymphoma in nonhuman primates after retroviral mediated gene transfer. J Exp Med 176: 1125 – 1135PubMedCrossRefGoogle Scholar
  11. Finter NB, Garland AJM, Telling RC (1990) Large scale mammalian cell culture: a perspective. In: Lubiniecki AS (ed) Large Scale Mammalaian Cell Culture Technology, Marcel Dekker New York, pp 1 – 14Google Scholar
  12. Griffiths B (1991) Cultural revolutions. Chem Ind, pp 682 – 684Google Scholar
  13. Gunter K, Khan AS, Noguchi PD (1993) The safety of retroviral vectors. Hum Gene Ther 4: 643 – 645PubMedCrossRefGoogle Scholar
  14. Kohn DB, Weinberg KI, Nolta JA, Heiss LN et al. (1995) Engraftment of gene modified umbilical cord blood cells in neonates with adenosine deaminase deficiency. Nat Med 10: 1017 – 1023CrossRefGoogle Scholar
  15. Kotani H, Newton PB, Zhang S, Chiang YL, Otto E, Weaver L, Blaese RM, Anderson WF, McGarrity G J (1994) Improved methods of retroviral vector transduction and production for gene therapy. Hum Gene Ther 5: 19 – 28PubMedCrossRefGoogle Scholar
  16. Long Z, Li L-P, Grooms T, Lockey C, Nader K, Mychkovsky M, Mueller S, Burimski, Ryan P, Kikuchi G, Ennist D, Marcus S, Otto E, McGarrity G (1998) Biosafety monitoring of patients receiving intracerebral injections of retroviral vector producer cells. Human Gene Ther 9: 1165 – 1172CrossRefGoogle Scholar
  17. Lubiniecki AS (1990) Continuous cell substrate considerations. In: Lubiniecki AS (ed) Large Scale Mammalian Cell Culture Technology. Marcel Dekker, New York, pp 495 – 513Google Scholar
  18. Lyons R, Forry-Schaudies S, Otto E et al. (1995) An improved vector encoding the herpes simplex thymidine kinase gene increases antitumor efficacy in vivo. Cancer Gene Ther 2: 273 – 280PubMedGoogle Scholar
  19. Markowitz D, Goff S, Bank A (1988) A safe packaging Une for gene transfer: separating viral genes on two different plasmids. J Virol 62: 1120 – 1124PubMedGoogle Scholar
  20. Martineau D, Klump WM, McCormack JE, DePolo NJ, Kamantigue E, Petrowski M, Hanlon J, Jolly D, Mento SJ, Sajjadi N (1997) Evaluation of PCR and ELISA assays for screening clinical trial subjects for replication competent retrovirus. Hum Gene Ther 8: 1231 – 1241PubMedCrossRefGoogle Scholar
  21. McCormack JE, Martineau D, DePolo N, Maifert S,Akbarian L, Townsend K, Lee W, Irwin M, Sajjadi N, Jolly DJ, Warner J (1997) Anti-vector immunoglobulin induced by retroviral vectors. Hum Gene Ther 8: 1263 – 1273Google Scholar
  22. McGarrity G J, Hoerner CL (1995) Biological safety in the biotechnoogy industry. In: Fleming DO, Richardson JH, Tulis JJ, Vesley D (eds) Laboratory safety, principles and practices, 2nd edn American Society for Microbiology, Washington DC, pp 119 – 132Google Scholar
  23. McGarrity GJ, Chiang YL (1997) Gene therapy of local tumors. In: MT Lotze and JT Rubin (eds) Regional therapy of advanced cancer, Lippincott-Raven, Philadelphia, pp 375 – 390Google Scholar
  24. Mento S J (1994) Status report on the development of retro vector gene transfer products. Process Development Issues in viral product manufacturing, The Williamsburg Bioprocessing ConferenceGoogle Scholar
  25. Miller AD, Buttimore C (1986) Redesign of retrovirus packaging cell lines to avoid recombination leading to helper virus production. Mol Cell Biol 6: 2895 – 2902PubMedGoogle Scholar
  26. National Institutes of Health (1997) Guidelines for research involving recombinant DNA molecules (NIH Guidelines) National Institutes of Health, Bethesda MD Federal Register January 31, 1997 (62 FR 4782)Google Scholar
  27. Otto E, Jones-Trower A, Vanin EF, Stambaugh K, Mueller SN, Anderson WF, McGarrity G J (1994) Characterization of a replication competent retrovirus resulting from recombination of packaging and vector sequences. Hum Gene Ther 5: 567 – 575PubMedCrossRefGoogle Scholar
  28. Rigg RJ, Chen J, Dando JS, Forestell SP, Plavec I, Bohnlein E (1996) A novel human amphotropic packaging cell line: high titer, complement resistance, and improved safety. Virology 218: 290 – 295PubMedCrossRefGoogle Scholar
  29. Smiley WR, Lambert B, Howard BD, Ibang C, Fong TC, Summers WS, Burrows FJ (1997) Establishment of parameters for optimal transduction efficiency and antitumor effects with purified high titer HSV-thymidine kinase retroviral vector in established solid tumors. Hum Gene Ther. 20: 965 – 977CrossRefGoogle Scholar
  30. Smith KT, Shepherd AJ, Boyd JE, Lees GM (1996) Gene delivery systems for use in gene therapy: an overview of quality assurance and safety issues. Gene Ther 3: 190 – 200PubMedGoogle Scholar
  31. Wiebe ME, May LH (1990) Cell banking. In: Lubiniecki AS (ed) Large scale mammalian cell culture technology, Marcel Dekker New York, pp 147 – 160Google Scholar
  32. Wilson CA, Ng T-H, Miller AE (1997). Evaluation of recommendations for replication-competent retrovirus testing associated with use of retroviral vectors. Hum Gene Ther 8: 869 – 874PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1999

Authors and Affiliations

  • H. Kotani
  • G. J. Mcgarrity

There are no affiliations available

Personalised recommendations