title: Immunogenicity of an adeno-vector vaccine expressing the F protein of a respiratory syncytial virus manufactured from serum-free suspension culture.
abstract: We have developed an efficient cell culture process to scale up the production of a recombinant adenovirus that expresses the membrane-trunked fusion protein of respiratory syncytial virus (RSV; Ad-F0ΔTM). Adherent cells of human embryonic kidney (HEK) 293-derived cell, 293A, which supports the production of E1/E3-deleted Ad-F0ΔTM when cultured in the presence of fetal bovine serum (FBS), were adapted to suspension growth under serum-free medium. In doing so, we studied the immunogenicity of Ad-F0ΔTMsus, which propagated in a bioreactor that was cultured with serum-free suspension of 293A, in comparison with Ad-F0ΔTMadh, which was produced from parental 293A cells that were adherently cultured in medium containing FBS. The size and morphology of Ad-F0ΔTMsus and Ad-F0ΔTMadh virions were identical upon inspection with electron microscopy. The results showed that anti-F IgG and RSV-neutralizing titer were raised in the serum of both mice that were intranasally immunized twice with Ad-F0ΔTMsus or Ad-F0ΔTMadh at two-week injection intervals. Furthermore, the immune responses persisted for six months after vaccination. Activation of F protein-specific CD8(+) T cell's epitope associated IFN-ɣ and IL-4 was induced in both Ad-F0ΔTMsus- and Ad-F0ΔTMadh, but not in Ad-LacZsus, -immunized mouse splenocytes. No vaccine-enhanced lung inflammation, airway mucus occlusion or eosinophils infiltration were observed in Ad-immunized mice followed by RSV challenge; however, these symptoms were observed following immunization with formalin-inactivated RSV vaccine. These results indicate that the safety and potency of Ad-F0ΔTM produced from either adherent cells or suspension and serum-free cells are the same.
authors.affiliation: Institute of Infectious Disease and Vaccinology, National Health Research Institutes, Zhunan Town, Miaoli County, Taiwan; Graduate Program of Biotechnology in Medicine, Institute of Molecular and Cellular Biology, National Tsing Hua University, Hsinchu 300, Taiwan.
authors.forename: Hsiao-Yun
authors.initials: HY
authors.lastname: Shao
authors.affiliation: Institute of Infectious Disease and Vaccinology, National Health Research Institutes, Zhunan Town, Miaoli County, Taiwan.
authors.forename: Huai-Sheng
authors.initials: HS
authors.lastname: Hsu
authors.affiliation: Institute of Infectious Disease and Vaccinology, National Health Research Institutes, Zhunan Town, Miaoli County, Taiwan.
authors.forename: Shu-Ling
authors.initials: SL
authors.lastname: Yu
authors.affiliation: Institute of Oral Medicine, National Cheng Kung University College of Medicine and Hospital, Tainan 701, Taiwan.
authors.forename: Shang-Rung
authors.initials: SR
authors.lastname: Wu
authors.affiliation: Institute of Infectious Disease and Vaccinology, National Health Research Institutes, Zhunan Town, Miaoli County, Taiwan.
authors.forename: Kai-Chieh
authors.initials: KC
authors.lastname: Hu
authors.affiliation: Institute of Infectious Disease and Vaccinology, National Health Research Institutes, Zhunan Town, Miaoli County, Taiwan; Graduate School of Life Science, National Defense Medical Center, Taipei 114, Taiwan.
authors.forename: Ching-Kun
authors.initials: CK
authors.lastname: Chang
authors.affiliation: Institute of Infectious Disease and Vaccinology, National Health Research Institutes, Zhunan Town, Miaoli County, Taiwan.
authors.forename: Chia-Chyi
authors.initials: CC
authors.lastname: Liu
authors.affiliation: Institute of Infectious Disease and Vaccinology, National Health Research Institutes, Zhunan Town, Miaoli County, Taiwan; Graduate Institute of Immunology, China Medical University, Taichung, Taiwan. Electronic address: choeyenh@nhri.org.tw.
authors.forename: Yen-Hung
authors.initials: YH
authors.lastname: Chow
mesh_terms: D000256:Adenoviridae / Q000235:genetics* / Q000276:immunology; D000818:Animals; D057134:Antibodies, Neutralizing / Q000097:blood / Q000276:immunology; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D018929:Cell Culture Techniques; D002460:Cell Line; D016895:Culture Media, Serum-Free; D004195:Disease Models, Animal; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics* / Q000276:immunology; D057809:HEK293 Cells; D006801:Humans; D007117:Immunization, Secondary; D051379:Mice; D009500:Neutralization Tests; D051381:Rats; D018357:Respiratory Syncytial Virus Infections / Q000276:immunology / Q000517:prevention & control; D022261:Respiratory Syncytial Virus Vaccines / Q000235:genetics / Q000276:immunology*; D012136:Respiratory Syncytial Viruses / Q000235:genetics* / Q000276:immunology*; D015164:Vaccines, Inactivated / Q000276:immunology; D014760:Viral Fusion Proteins / Q000235:genetics* / Q000276:immunology*
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D016895:Culture Media, Serum-Free; C526850:F protein, human respiratory syncytial virus; D022261:Respiratory Syncytial Virus Vaccines; D015164:Vaccines, Inactivated; D014760:Viral Fusion Proteins
keywords: Adenovirus; Immunogenicity; Respiratory syncytial virus; Vaccine