title: Development of a full-length cDNA-derived enterovirus A71 vaccine candidate using reverse genetics technology.
abstract: Enterovirus A71 (EV-A71) is responsible for epidemics of hand, foot and mouth disease (HFMD) in young children. To circumvent difficulties in obtaining clinical enterovirus isolates that might be contaminated with other viruses, a platform technology was developed to quickly generate vaccine virus strains based on the published enterovirus genomic sequences. A recombinant plasmid containing the full-length infectious cDNA clone of EV-A71 vaccine strain E59 was directly generated after transfecting the recombinant plasmid into Vero, RD or HEK293A cells, and phenotypic characteristics similar to the parental strain were observed. The cDNA-derived infectious EV-A71 virus grown in Vero cells produced relatively stable virus titers in both T-flasks and microcarrier culture systems. To evaluate the genetic stability of the cDNA-derived EV-A71 viruses, the immunodominant structural proteins, VP1 and VP2, of the recombinant EV-A71 viruses were sequenced and analyzed. The cDNA-derived EV-A71 virus showed weak pathogenicity in a human SCARB2 mouse model. These results show the successful generation of a recombinant virus derived from a published viral genomic sequence that demonstrated good genetic stability and viral yields, which could represent an efficient and safe vaccine strain for cGMP-grade manufacturing.
authors.affiliation: National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Zhunan Town, Miaoli County, Taiwan.
authors.forename: Ya-Ting
authors.initials: YT
authors.lastname: Yang
authors.affiliation: National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Zhunan Town, Miaoli County, Taiwan; Graduate Institute of Immunology, China Medical University, Taichung, Taiwan.
authors.forename: Yen-Hung
authors.initials: YH
authors.lastname: Chow
authors.affiliation: National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Zhunan Town, Miaoli County, Taiwan.
authors.forename: Kuang-Nan
authors.initials: KN
authors.lastname: Hsiao
authors.affiliation: National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Zhunan Town, Miaoli County, Taiwan.
authors.forename: Kai-Chieh
authors.initials: KC
authors.lastname: Hu
authors.affiliation: Centers for Disease Control, Taipei, Taiwan.
authors.forename: Jen-Ron
authors.initials: JR
authors.lastname: Chiang
authors.affiliation: Institute of Biotechnology, Department of Life Science, National Tsing Hua University, Hsinchu, Taiwan.
authors.forename: Suh-Chin
authors.initials: SC
authors.lastname: Wu
authors.affiliation: National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Zhunan Town, Miaoli County, Taiwan; Graduate Institute of Immunology, China Medical University, Taichung, Taiwan; Institute of Biotechnology, Department of Life Science, National Tsing Hua University, Hsinchu, Taiwan.
authors.forename: Pele
authors.initials: P
authors.lastname: Chong
authors.affiliation: National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Zhunan Town, Miaoli County, Taiwan. Electronic address: 010113@nhri.org.tw.
authors.forename: Chia-Chyi
authors.initials: CC
authors.lastname: Liu
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D057134:Antibodies, Neutralizing / Q000097:blood / Q000276:immunology; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology; D002522:Chlorocebus aethiops; D018076:DNA, Complementary; D004195:Disease Models, Animal; D029821:Enterovirus A, Human / Q000235:genetics* / Q000276:immunology* / Q000472:pathogenicity; D004769:Enterovirus Infections / Q000276:immunology* / Q000517:prevention & control / Q000821:virology; D023061:Gene Order; D016679:Genome, Viral; D042822:Genomic Instability; D006801:Humans; D016056:Immunodominant Epitopes / Q000235:genetics / Q000276:immunology; D051379:Mice; D008822:Mice, Transgenic; D009154:Mutation; D019444:Vaccines, DNA / Q000235:genetics* / Q000276:immunology*; D014709:Vero Cells; D014779:Virus Replication
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D000956:Antigens, Viral; D018076:DNA, Complementary; D016056:Immunodominant Epitopes; D019444:Vaccines, DNA
keywords: Enterovirus A71 (EV-A71); Full-length infectious clone; Microcarrier; Vaccine; cDNA-derived virus