Efficacy of adenoviral vectors (AdV) was checked by transduction of six different cell lines (COS-7, HUVEC, HMEC-1, NIH 3T3, HaCaT, and B16(F10)) with the vectors containing reporter gene beta-galactosidase (Adbeta-gal). We optimised the adsorption time and dose of Ad beta-gal. The transduction with efficacy of up to 100% was obtained for the doses 10-100 IU/cell. In order to examine the effect of transduction procedure on biology of the cells, we measured the production of major proinflammatory cytokines. In several cell lines (HMEC-1, HUVEC, HaCaT), we found the induction of IL-6 synthesis and decrease in the cell viability, particularly in the case of the highest Adbeta-gal dose. However, the treatment with adenoviral vectors did not induce TNF generation and did not modify proliferation of cells.
The paper presents the production of adenoviral vectors (AdV) containing beta-galactosidase (Adbeta-gal), from the transfer of recombinant viral DNA into packing cell line (HEK293) to the titration of viral particles. Optimisation of the methods (preparation of DNA for transfection, adsorption time during the infection of cells, amount of serum in the medium, time-point of vector isolation) enables obtaining a titer of up to 1010 IU/mL. Adbeta-gal were titrated with several methods, with beta-gal in situ staining used as a reference. We found that the most suitable titration method of the vectors containing other than reporter genes was the Rapid Titer ELISA kit?.
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