Abstract
The use of chimeric pseudotyped vectors is a common way to modify the adenoviral tropism by replacing the fiber protein. In this chapter the procedure to generate a chimeric adenovirus pre-stock from a plasmid containing the adenoviral genome is described. Also, the chimeric adenovirus replicative cycle to increase the yield in further productions is determined. Finally, two different protocols, in culture plates and in suspension cultures, to produce the virus at large scale are also detailed.
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References
Kojaoghlanian T, Flomenberg P, Horwitz MS (2003) The impact of adenovirus infection on the immunocompromised host. Rev Med Virol 13:155–171
Haddada H, Cordier L, Perricaudet M (1995) Gene therapy using adenovirus vectors. Curr Top Microbiol Immunol 199(Pt 3):297–306
Wivel NA, Gao G, Wilson JM (1999) Adenovirus vectors. The development of human gene therapy. Cold Spring Harbor monograph series, pp 87–110
Thomas CE, Ehrhardt A, Kay MA (2003) Progress and problems with the use of viral vectors for gene therapy. Nat Rev Genet 4:346–358
Verma IM, Weitzman MD (2005) Gene therapy: twenty-first century medicine. Annu Rev Biochem 74:711–738
Gall J, Kass-Eisler A, Leinwand L, Falck-Pedersen E (1996) Adenovirus type 5 and 7 capsid chimera: fiber replacement alters receptor tropism without affecting primary immune neutralization epitopes. J Virol 70:2116–2123
Nakamura T, Sato K, Hamada H (2003) Reduction of natural adenovirus tropism to the liver by both ablation of fiber-coxsackievirus and adenovirus receptor interaction and use of replaceable short fiber. J Virol 77:2512–2521
Roberts DM, Nanda A, Havenga MJ, Abbink P, Lynch DM, Ewald BA et al (2006) Hexon-chimaeric adenovirus serotype 5 vectors circumvent pre-existing anti-vector immunity. Nature 441:239–243
Miralles M, Segura MM, Puig M, Bosch A, Chillon M (2012) Efficient amplification of chimeric adenovirus 5/40S vectors carrying the short fiber protein of Ad40 in suspension cell cultures. PLoS One 7:e42073
Sherwood V, Burgert HG, Chen YH, Sanghera S, Katafigiotis S, Randall RE et al (2007) Improved growth of enteric adenovirus type 40 in a modified cell line that can no longer respond to interferon stimulation. J Gen Virol 88:71–76
Tiemessen CT, Kidd AH (1994) Adenovirus type 40 and 41 growth in vitro: host range diversity reflected by differences in patterns of DNA replication. J Virol 68:1239–1244
Rodriguez E, Romero C, Ferrer M, Burgueño JF, Rio A, Gil E, Hamada H, Bosch A, Gasull MA, Fernandez E, Chillon M (2006) Therapeutic potential of the Chimeric Adenovirus 5/40 as a vector for intestine directed gene therapy. Mol Ther 13:S5
Arcasoy SM, Latoche JD, Gondor M, Pitt BR, Pilewski JM (1997) Polycations increase the efficiency of adenovirus-mediated gene transfer to epithelial and endothelial cells in vitro. Gene Ther 4:32–38
Jacobsen F, Hirsch T, Mittler D, Schulte M, Lehnhardt M, Druecke D et al (2006) Polybrene improves transfection efficacy of recombinant replication-deficient adenovirus in cutaneous cells and burned skin. J Gene Med 8:138–146
Geisse S, Di Maiuta N, Ten Buren B, Henke M (2005) The secrets of transfection in serum-free suspension culture. In: Gòdia F, Fussenegger M (eds) Animal cell technology meets genomics. ESACT Proceedings. Springer, Netherlands, pp 373–376
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Miralles, M., Garcia, M., Tejero, M., Bosch, A., Chillón, M. (2014). Production of Chimeric Adenovirus. In: Chillón, M., Bosch, A. (eds) Adenovirus. Methods in Molecular Biology, vol 1089. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-679-5_16
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DOI: https://doi.org/10.1007/978-1-62703-679-5_16
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