Novel replication-incompetent adenoviral B-group vectors: high vector stability and yield in PER.C6 cells
Havenga, M. and Vogels, R. and Zuijdgeest, D. and Radosevic, K. and Mueller, S. and Sieuwerts, M. and Weichold, F. and Damen, I. and Kaspers, J. and Lemckert, A. and van Meerendonk, M. and van der Vlugt, R. and Holterman, L. and Hone, D. and Skeiky, Y. and Mintardjo, R. and Gillissen, G. and Barouch, D. and Sadoff, J. and Goudsmit, J.,, 87, 2135-2143 (2006),
doi = https://doi.org/10.1099/vir.0.81956-0,
publicationName = Microbiology Society,
issn = 0022-1317,
abstract= Adenoviral vectors based on adenovirus type 35 (rAd35) have the advantage of low natural vector immunity and induce strong, insert-specific T- and B-cell responses, making them prime-candidate vaccine carriers. However, severe vector-genome instability of E1-deleted rAd35 vectors was observed, hampering universal use. The instability of E1-deleted rAd35 vector proved to be caused by low pIX expression induced by removal of the pIX promoter, which was located in the E1B region of B-group viruses. Reinsertion of a minimal pIX promoter resulted in stable vectors able to harbour large DNA inserts (>5 kb). In addition, it is shown that replacement of the E4-Orf6 region of Ad35 by the E4-Orf6 region of Ad5 resulted in successful propagation of an E1-deleted rAd35 vector on existing E1-complementing cell lines, such as PER.C6 cells. The ability to produce these carriers on PER.C6 contributes significantly to the scale of manufacturing of rAd35-based vaccines. Next, a stable rAd35 vaccine was generated carrying Mycobacterium tuberculosis antigens Ag85A, Ag85B and TB10.4. The antigens were fused directly, resulting in expression of a single polyprotein. This vaccine induced dose-dependent CD4+ and CD8+ T-cell responses against multiple antigens in mice. It is concluded that the described improvements to the rAd35 vector contribute significantly to the further development of rAd35 carriers for mass-vaccination programmes for diseases such as tuberculosis, AIDS and malaria.,
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