dengue type 4 live-attenuated vaccine viruses passaged in vero cells affect genetic stability and dengue-induced hemorrhaging in mice登革4型减毒活疫苗疫苗病毒通过州立细胞影响小鼠的遗传稳定性和dengue-induced大出血.pdfVIP

Dengue Type 4 Live-Attenuated Vaccine Viruses Passaged in Vero Cells Affect Genetic Stability and Dengue-Induced Hemorrhaging in Mice 1 3 3 3 1,2 Hsiang-Chi Lee , Yu-Ting Yen , Wen-Yu Chen , Betty A. Wu-Hsieh , Suh-Chin Wu * 1 Institute of Biotechnology, Department of Life Science, National Tsing Hua University, Hsinchu, Taiwan, 2 National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Miaoli, Taiwan, 3 Graduate Institute of Immunology, National Taiwan University, College of Medicine, Taipei, Taiwan Abstract Most live-attenuated tetravalent dengue virus vaccines in current clinical trials are produced from Vero cells. In a previous study we demonstrated that an infectious cDNA clone-derived dengue type 4 (DEN-4) virus retains higher genetic stability in MRC-5 cells than in Vero cells. For this study we investigated two DEN-4 viruses: the infectious cDNA clone-derived DEN-4 2A and its derived 39 NCR 30-nucleotide deletion mutant DEN-4 2AD30, a vaccine candidate. Mutations in the C-prM-E, NS2B-NS3, and NS4B-NS5 regions of the DEN genome were sequenced and compared following cell passages in Vero and MRC-5 cells. Our results indicate stronger genetic stability in both viruses following MRC-5 cell passages, leading to significantly lower RNA polymerase error rates when the DEN-4 virus is used for genome replication. Although no significant increases in virus titers were observed following cell passages, DEN-4 2A and DEN-4 2AD30 virus titers following Vero cell passages were 17-fold to 25-fold higher than titers following MRC-5 cell passages. Neurovirulence for DEN-4 2A and DEN-4 2AD30 viruses increased significantly following passages in Vero