Abstract
Adenoviruses are promising vectors for gene therapy. The production of adenoviral vectors (AdV), however, is limited by the cell density effect, namely when cell infection is performed at above 106 cells/ml, a drop in cell-specific adenovirus productivity occurs. Our results also show that the coxsackie adenovirus receptor (CAR) plays an important role in AdV production. CAR expression of infected cells varied with culture time and the cell-specific AdV productivity dropped rapidly along with decreased CAR expression. Furthermore, CAR expression of cells was maintained at a high level by replacing the medium or supplementing it with trichostatin A, which could improve the cell-specific productivity. Thus, a higher CAR expression level at infection time could improve cell-specific AdV productivity at high cell densities.
Similar content being viewed by others
References
Coyne CB, Bergelson JM (2005) CAR: a virus receptor within the tight junction. Adv Drug Deliv Rev 57:869–882
Ferreira TB, Ferreira AL, Carrondo MJT, Alves PM (2005a) Effect of reefed strategies and non-ammoniagenic medium on adenovirus production at high cell densities. J Biotechnol 119:272–280
Ferreira TB, Ferreira AL, Carrondo MJT, Alves PM (2005b) Two different serum free media and osmolality effect upon 293 cell growth and adenovirus production. Biotechnol Lett 27:1809–1813
Garnier A, Cote J, Nadeau I, Kamen A, Massie B (1994) Scale-up of the adenovirus expression system for the production of recombinant protein in human 293S cells. Cytotechnology 15:145–155
Henry O, Dormond E, Perrier M, Kamen A (2004) Insights into adenoviral vector production kinetics in acoustic filter-based perfusion cultures. Biotechnol Bioeng 86:765–774
Jardon M, Garnier A (2003) pH, pCO2, and temperature effect on R-adenovirus production. Biotechnol Prog 19:202–208
Kamen A, Henry O (2004) Development and optimization of an adenovirus production process. J Gen Med 6:184–192
Kauwe LKM (2003) Endocytosis of adenovirus and adenovirus capsid proteins. Adv Drug Deliv Rev 55:1485–1496
Kim M, Zinn KR, Barnett BG, Sumerel LA, Krasnykh V, Curiel DT, Douglas JT (2002) The therapeutic efficacy of adenoviral vectors for cancer gene therapy is limited by a low level of primary adenovirus receptors on tumour cells. Eur J Cancer 38:1917–1926
Law LK, Davidson BL (2005) What does it take to bind CAR? Mol Ther 12:599–609
Matsumoto K, Shariat SF, Ayala GE, Rauen KA, Lerner SP (2005) Loss of coxsackie and adenovirus receptor expression is associated with features of aggressive bladder cancer. Urology 66:441–446
Nadeau I, Kamen A (2003) Production of adenovirus vector for gene therapy. Biotechnol Adv 20:475–489
Nadeau I, Garnier A, Cote J, Massie B, Chavarie C, Kamen A (1996) Improvement of recombinant protein production with human adenovirus/293S expression system using fed-batch strategies. Biotechnol Bioeng 51:613–623
Nagel H, Maag S, Tassis A, Nestle FO, Greber UF, Hemmi S (2003) The αvβ5 integrin of hematopoietic and nonheatopoietic cells is a transduction receptor of RGD-4C fiber-modified adenoviruses. Gene Ther 10:1643–1653
Nemerow GR (2000) Cell receptors involved in adenovirus entry. Virology 274:1–4
Reichert JC, Rosensweig CJ, Faden LB, Dewitz MC (2005) Monoclonal antibody successes in the clinic. Nat Biotechnol 23:1073–1078
Seidman MA, Hogan SM, Wendland RL, Worgall S, Crystal RG, Leopold PL (2001) Variation in adenovirus receptor expression and adenovirus vector-mediated transgene expression at defined stages of the cell cycle. Mol Ther 4:13–21
Zhang C, Ferreira TB, Cruz PE, Alves PM, Haury M, Carrondo MJT (2006) The importance of 293 cell cycle phase on adenovirus vector production. Enzyme Microb Technol 39:1328–1332
Acknowledgement
This work was partially funded by the National Science Foundation of China (NSFC projects no. 20706016).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Liu, X., Wang, Y., Niu, H. et al. The improvement of adenovirus vector production by increased expression of coxsackie adenovirus receptor. Biotechnol Lett 31, 939–944 (2009). https://doi.org/10.1007/s10529-009-9971-y
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10529-009-9971-y