Inactivated Enterovirus 71 Vaccine Produced by 200-L Scale Serum-Free Microcarrier Bioreactor System Provides Cross-Protective Efficacy in Human SCARB2 Transgenic Mouse.
Epidemics and outbreaks caused by infections of several subgenotypes of EV71 and other serotypes of coxsackie A viruses have raised serious public health concerns in the Asia-Pacific region. These concerns highlight the urgent need to develop a scalable manufacturing platform for producing an effective and sufficient quantity of vaccines against deadly enteroviruses. In this report, we present a platform for the large-scale production of a vaccine based on the inactivated EV71(E59-B4) virus. The viruses were produced in Vero cells in a 200 L bioreactor with serum-free medium, and the viral titer reached 10(7) TCID50/mL 10 days after infection when using an MOI of 10(-4). The EV71 virus particles were harvested and purified by sucrose density gradient centrifugation. Fractions containing viral particles were pooled based on ELISA and SDS-PAGE. TEM was used to characterize the morphologies of the viral particles. To evaluate the cross-protective efficacy of the EV71 vaccine, the pooled antigens were combined with squalene-based adjuvant (AddaVAX) or aluminum phosphate (AlPO4) and tested in human SCARB2 transgenic (Tg) mice. The Tg mice immunized with either the AddaVAX- or AlPO4-adjuvanted EV71 vaccine were fully protected from challenges by the subgenotype C2 and C4 viruses, and surviving animals did not show any degree of neurological paralysis symptoms or muscle damage. Vaccine treatments significantly reduced virus antigen presented in the central nervous system of Tg mice and alleviated the virus-associated inflammatory response. These results strongly suggest that this preparation results in an efficacious vaccine and that the microcarrier/bioreactor platform offers a superior alternative to the previously described roller-bottle system.
Adimmune Corporation, Taichung, Taiwan.
Chia-Ying
CY
Wu
Institute of Infectious Disease and Vaccinology, National Health Research Institutes, Zhunan, Miaoli County, Taiwan.
Yi-Wen
YW
Lin
Adimmune Corporation, Taichung, Taiwan.
Chia-Ho
CH
Kuo
Adimmune Corporation, Taichung, Taiwan.
Wan-Hsin
WH
Liu
Adimmune Corporation, Taichung, Taiwan.
Hsiu-Fen
HF
Tai
Adimmune Corporation, Taichung, Taiwan.
Chien-Hung
CH
Pan
Adimmune Corporation, Taichung, Taiwan.
Yung-Tsung
YT
Chen
Agricultural Biotechnology Research Center, Academia Sinica, Taipei, Taiwan.
Pei-Wen
PW
Hsiao
Adimmune Corporation, Taichung, Taiwan.
Chi-Hsien
CH
Chan
Adimmune Corporation, Taichung, Taiwan.
Ching-Chuan
CC
Chang
Adimmune Corporation, Taichung, Taiwan; Enimmune Corporation, Taichung, Taiwan.
Chung-Cheng
CC
Liu
Institute of Infectious Disease and Vaccinology, National Health Research Institutes, Zhunan, Miaoli County, Taiwan; Graduate Institute of Immunology, China Medical University, Taichung, Taiwan.
Yen-Hung
YH
Chow
Adimmune Corporation, Taichung, Taiwan.
Juine-Ruey
JR
Chen
D000818:Animals; D019149:Bioreactors; D018925:Chemokines / Q000235:genetics; D002522:Chlorocebus aethiops; D003384:Coxsackievirus Infections / Q000235:genetics / Q000276:immunology / Q000517:prevention & control; D003429:Cross Reactions; D016207:Cytokines / Q000235:genetics; D029821:Enterovirus A, Human / Q000276:immunology*; D015870:Gene Expression; D006801:Humans; D051907:Lysosomal Membrane Proteins / Q000235:genetics* / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008822:Mice, Transgenic; D011817:Rabbits; D051116:Receptors, Scavenger / Q000235:genetics* / Q000276:immunology*; D015164:Vaccines, Inactivated / Q000096:biosynthesis / Q000276:immunology; D014709:Vero Cells; D014765:Viral Vaccines / Q000096:biosynthesis / Q000276:immunology*; D014776:Virus Cultivation
D018925:Chemokines; D016207:Cytokines; D051907:Lysosomal Membrane Proteins; D051116:Receptors, Scavenger; C074299:SCARB2 protein, human; D015164:Vaccines, Inactivated; D014765:Viral Vaccines