1887

Abstract

The present work studies the physico-chemical properties of retroviral vector membrane, in order to provide some explanation for the inactivation kinetics of these vectors and to devise new ways of improving transduction efficiency. For this purpose, vectors with an amphotropic envelope produced by TE Fly A7 cells at two culture temperatures (37 and 32 °C) were characterized by different techniques. Electron paramagnetic resonance (EPR) results showed that vectors produced at 32 °C are more rigid than those produced at 37 °C. Further characterization of vector membrane composition allowed us to conclude that the vector inactivation rate increases with elevated cholesterol to phospholipid ratio. Differential scanning calorimetry (DSC) showed that production temperature also affects the conformation of the membrane proteins. Transduction studies using HCT116 cells and tri-dimensional organ cultures of mouse skin showed that vectors produced at 37 °C have higher stability and thus higher transduction efficiency in gene therapy relevant cells as compared with vectors produced at 32 °C. Overall, vectors produced at 37 °C show an increased stability at temperatures below 4 °C. Since vector membrane physico-chemical properties are affected in response to changes in culture temperature, such changes, along with alterations in medium composition, can be used prospectively to improve the stability and the transduction efficiency of retroviral vectors for therapeutic purposes.

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2006-05-01
2020-01-29
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References

  1. Aloia, R. C., Jensen, F. C., Curtain, C. C., Mobley, P. W. & Gordon, L. M. ( 1988; ). Lipid composition and fluidity of the human immunodeficiency virus. Proc Natl Acad Sci U S A 85, 900–904.[CrossRef]
    [Google Scholar]
  2. Aloia, R. C., Tian, H. & Jensen, F. C. ( 1993; ). Lipid composition and fluidity of the human immunodeficiency virus envelope and host cell plasma membranes. Proc Natl Acad Sci U S A 90, 5181–5185.[CrossRef]
    [Google Scholar]
  3. Beer, C., Meyer, A., Muller, K. & Wirth, M. ( 2003; ). The temperature stability of mouse retroviruses depends on the cholesterol levels of viral lipid shell and cellular plasma membrane. Virology 308, 137–146.[CrossRef]
    [Google Scholar]
  4. Brown, D. A. & London, E. ( 2000; ). Structure and function of sphingolipid- and cholesterol-rich membrane rafts. J Biol Chem 275, 17221–17224.[CrossRef]
    [Google Scholar]
  5. Carmo, M., Peixoto, C., Coroadinha, A. S., Alves, P. M., Cruz, P. E. & Carrondo, M. J. ( 2004; ). Quantitation of MLV-based retroviral vectors using real-time RT-PCR. J Virol Methods 119, 115–119.[CrossRef]
    [Google Scholar]
  6. Cruz, P. E., Almeida, J. S., Murphy, P. N., Moreira, J. L. & Carrondo, M. J. ( 2000; ). Modeling retrovirus production for gene therapy. 1. Determination of optimal bioreaction mode and harvest strategy. Biotechnol Prog 16, 213–221.[CrossRef]
    [Google Scholar]
  7. Del Rio, M., Gache, Y., Jorcano, J. L., Meneguzzy, G. & Larcher, F. ( 2004; ). Current approaches and perspectives in human keratinocyte-based gene therapies. Gene Ther 11, S57–S63.[CrossRef]
    [Google Scholar]
  8. Gény-Fiamma, C., Millot, L., Rocca, C., Danos, O. & Merten, O. W. ( 2004; ). Optimization of the production of retroviral vectors. Influences of the sugar source. In Animal Cell Technology. Basic & Applied Aspects, pp. 89–97. Edited by K. Yagasaki. Dordrecht, NL: Kluwer Academic Publishers.
  9. Gershonowitz, A., Itach, E. G., Shouval, D., Mitrani, D., Ilan, Y. & Mitrani, E. ( 2004; ). Development of a scaled up liver device incorporating cryo-preserved pig liver micro-organs. J Hepatol 41, 950–956.[CrossRef]
    [Google Scholar]
  10. Gordon, L. M., Jensen, F. C., Curtain, C. C., Mobley, P. W. & Aloia, R. C. ( 1988; ). Thermotropic lipid phase separation in the human immunodeficiency virus. Biochim Biophys Acta 943, 331–342.[CrossRef]
    [Google Scholar]
  11. Hammarstedt, M., Wallengren, K., Pedersen, K. W., Roos, N. & Garoff, H. ( 2000; ). Minimal exclusion of plasma membrane proteins during retroviral envelope formation. Proc Natl Acad Sci U S A 97, 7527–7532.[CrossRef]
    [Google Scholar]
  12. Hasson, E., Slovatizky, Y., Shimoni, Y., Falk, H., Panet, A. & Mitrani, E. ( 2005; ). Solid tissues can be manipulated ex vivo and used as vehicles for gene therapy. J Gene Med 7, 926–935.[CrossRef]
    [Google Scholar]
  13. Higashikawa, F. & Chang, L. ( 2001; ). Kinetic analyses of stability of simple and complex retroviral vectors. Virology 280, 124–131.[CrossRef]
    [Google Scholar]
  14. Kjaer, S. & Ibanez, C. F. ( 2003; ). Intrinsic susceptibility to misfolding of a hot-spot for Hirschsprung disease mutations in the ectodomain of RET. Hum Mol Genet 12, 2133–2144.[CrossRef]
    [Google Scholar]
  15. Le Doux, J. M., Davis, H. E., Morgan, J. R. & Yarmush, M. L. ( 1999; ). Kinetics of retrovirus production and decay. Biotechnol Bioeng 63, 654–662.[CrossRef]
    [Google Scholar]
  16. Lee, G. M., Choi, J. H., Jun, S. C. & Palsson, B. O. ( 1998; ). Temperature significantly affects retroviral vector production and deactivation rates, and thereby determines retroviral titers. Bioprocess Eng 19, 343–349.[CrossRef]
    [Google Scholar]
  17. McTaggart, S. & Al-Rubeai, M. ( 2002; ). Retroviral vectors for human gene delivery. Biotechnol Adv 20, 1–31.[CrossRef]
    [Google Scholar]
  18. Mountain, A. ( 2000; ). Gene therapy: the first decade. Trends Biotechnol 18, 119–128.[CrossRef]
    [Google Scholar]
  19. Nguyen, D. H. & Hildreth, J. E. K. ( 2000; ). Evidence for budding of human immunodeficiency virus type 1 selectively from glycolipid-enriched membrane lipid rafts. J Virol 74, 3264–3272.[CrossRef]
    [Google Scholar]
  20. O'Reilly, L. & Roth, M. J. ( 2003; ). Identification of conformational and cold-sensitive mutations in the MuLV envelope protein. Virology 312, 337–349.[CrossRef]
    [Google Scholar]
  21. Pessin, J. E. & Glaser, M. ( 1980; ). Budding of Rous sarcoma virus and vesicular stomatitis virus from localized lipid regions in the plasma membrane of chicken embryo fibroblasts. J Biol Chem 255, 9044–9050.
    [Google Scholar]
  22. Pizzato, M., Merten, O. W., Blair, E. D. & Takeuchi, Y. ( 2001; ). Development of a suspension packaging cell line for production of high titre, serum-resistant murine leukemia virus vectors. Gene Ther 8, 737–745.[CrossRef]
    [Google Scholar]
  23. Rein, A., Yang, C., Haynes, J. A., Mirro, J. & Copans, R. W. ( 1998; ). Evidence for cooperation between murine leukemia virus env molecules in mixed oligomers. J Virol 72, 3432–3435.
    [Google Scholar]
  24. Rheinwald, J. D. & Green, H. ( 1975; ). Serial cultivation of strains of human epidermal keratinocytes: the formation of keratinizing colonies from single cells. Cell 6, 331–343.[CrossRef]
    [Google Scholar]
  25. Simons, K. & Ikonen, E. ( 1997; ). Functional rafts in cell membranes. Nature 387, 569–572.[CrossRef]
    [Google Scholar]
  26. Simons, K. & Toomre, D. ( 2000; ). Lipid rafts and signal transduction. Nat Rev Mol Cell Biol 1, 31–39.
    [Google Scholar]
  27. Slosberg, B. N. & Montelaro, R. C. ( 1982; ). A comparison of the mobilities and thermal transitions of retrovirus lipid envelopes and host cell plasma membranes by electron spin resonance spectroscopy. Biochim Biophys Acta 689, 393–402.[CrossRef]
    [Google Scholar]
  28. Zhao, Y., Lee, S. & Anderson, W. F. ( 1997; ). Functional interactions between monomers of the retroviral envelope protein complex. J Virol 71, 6967–6972.
    [Google Scholar]
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