Propagation and immunological characterization of coxsackievirus A10 in a serum-free HEK293A cell culture system

Sheng Chieh Lien; Yu Sheng Shen; Hsiao Yu Lin; Shang Rung Wu; Chih Yeu Fang; Chi Hsun Chen; Yi An Chen; Pele Choi Sing Chong; Ming Hsi Huang; Yen Hung Chow; Jen Ren Wang; Suh Chin Wu; Chia Chyi Liu

doi:10.1016/j.virusres.2023.199101

Propagation and immunological characterization of coxsackievirus A10 in a serum-free HEK293A cell culture system

Sheng Chieh Lien, Yu Sheng Shen, Hsiao Yu Lin, Shang Rung Wu, Chih Yeu Fang, Chi Hsun Chen, Yi An Chen, Pele Choi Sing Chong, Ming Hsi Huang, Yen Hung Chow, Jen Ren Wang, Suh Chin Wu, Chia Chyi Liu

Research output: Contribution to journal › Article › peer-review

2 Citations (Scopus)

Abstract

Coxsackievirus A10 (CVA10) is one of enteroviral pathogens that cause the hand, foot, and mouth disease (HFMD). Since CVA10 was reported to be not easily propagated in the Vero cell culture, a feasible manufacture process for producing formalin-inactivated CVA10 vaccine is urgently needed. Several cell lines that commonly used for viral vaccine production was tested for CVA10 (M2014 strain) culture in this study, and our result showed that CVA10 could be easily propagated in the HEK293A cells. A serum-free HEK293A cell culture system was developed for CVA10 production and the yields have reached over 10⁸ TCID₅₀/mL. The biochemical and immunogenic properties of CVA10 particles obtained from this serum-free HEK293A culture were identical to our previous study. Two major particles of CVA10 were separated by ultracentrifugation, and only the infectious mature particles were capable of inducing CVA10 neutralizing antibody responses in the mouse immunogenicity studies. Additionally, we found that coxsackievirus A6 and enterovirus A71 could also be easily propagated using this serum-free HEK293A cell culture system. Our results provide a solution to overcome the obstacle in the propagation of CVA10 and facilitate the development of multivalent vaccines for prevention of HFMD.

Original language	English
Article number	199101
Journal	Virus Research
Volume	329
DOIs	https://doi.org/10.1016/j.virusres.2023.199101
Publication status	Published - 2023 May

All Science Journal Classification (ASJC) codes

Virology
Infectious Diseases
Cancer Research

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.virusres.2023.199101

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Lien, S. C., Shen, Y. S., Lin, H. Y., Wu, S. R., Fang, C. Y., Chen, C. H., Chen, Y. A., Chong, P. C. S., Huang, M. H., Chow, Y. H., Wang, J. R., Wu, S. C., & Liu, C. C. (2023). Propagation and immunological characterization of coxsackievirus A10 in a serum-free HEK293A cell culture system. Virus Research, 329, Article 199101. https://doi.org/10.1016/j.virusres.2023.199101

@article{86f8f950309b49e794a8dc055e23e433,

title = "Propagation and immunological characterization of coxsackievirus A10 in a serum-free HEK293A cell culture system",

abstract = "Coxsackievirus A10 (CVA10) is one of enteroviral pathogens that cause the hand, foot, and mouth disease (HFMD). Since CVA10 was reported to be not easily propagated in the Vero cell culture, a feasible manufacture process for producing formalin-inactivated CVA10 vaccine is urgently needed. Several cell lines that commonly used for viral vaccine production was tested for CVA10 (M2014 strain) culture in this study, and our result showed that CVA10 could be easily propagated in the HEK293A cells. A serum-free HEK293A cell culture system was developed for CVA10 production and the yields have reached over 108 TCID50/mL. The biochemical and immunogenic properties of CVA10 particles obtained from this serum-free HEK293A culture were identical to our previous study. Two major particles of CVA10 were separated by ultracentrifugation, and only the infectious mature particles were capable of inducing CVA10 neutralizing antibody responses in the mouse immunogenicity studies. Additionally, we found that coxsackievirus A6 and enterovirus A71 could also be easily propagated using this serum-free HEK293A cell culture system. Our results provide a solution to overcome the obstacle in the propagation of CVA10 and facilitate the development of multivalent vaccines for prevention of HFMD.",

author = "Lien, {Sheng Chieh} and Shen, {Yu Sheng} and Lin, {Hsiao Yu} and Wu, {Shang Rung} and Fang, {Chih Yeu} and Chen, {Chi Hsun} and Chen, {Yi An} and Chong, {Pele Choi Sing} and Huang, {Ming Hsi} and Chow, {Yen Hung} and Wang, {Jen Ren} and Wu, {Suh Chin} and Liu, {Chia Chyi}",

note = "Funding Information: This work was supported by the Ministry of Health and Welfare, the Taiwan CDC and the National Health Research Institutes, Taiwan. We also thank the technical services provided by “the i-MANI center of the National Core Facility for Biopharmaceuticals, Ministry of Science and Technology, Taiwan” as well as the technical service from the Instrument Development Center of the National Cheng Kung University. Funding Information: This work was supported by the National Health Research Institutes , Taiwan [Grant Nos. 06A1-IVPP13–014 , 09A1-IVPP13–014 , 11A1-IVPP14–014 , 12A1-IVPP18–014 ]. Funding Information: This work was supported by the National Health Research Institutes, Taiwan [Grant Nos. 06A1-IVPP13–014, 09A1-IVPP13–014, 11A1-IVPP14–014, 12A1-IVPP18–014]. This work was supported by the Ministry of Health and Welfare, the Taiwan CDC and the National Health Research Institutes, Taiwan. We also thank the technical services provided by “the i-MANI center of the National Core Facility for Biopharmaceuticals, Ministry of Science and Technology, Taiwan” as well as the technical service from the Instrument Development Center of the National Cheng Kung University. Publisher Copyright: {\textcopyright} 2023 The Author(s)",

year = "2023",

month = may,

doi = "10.1016/j.virusres.2023.199101",

language = "English",

volume = "329",

journal = "Virus Research",

issn = "0168-1702",

publisher = "Elsevier",

}

TY - JOUR

T1 - Propagation and immunological characterization of coxsackievirus A10 in a serum-free HEK293A cell culture system

AU - Lien, Sheng Chieh

AU - Shen, Yu Sheng

AU - Lin, Hsiao Yu

AU - Wu, Shang Rung

AU - Fang, Chih Yeu

AU - Chen, Chi Hsun

AU - Chen, Yi An

AU - Chong, Pele Choi Sing

AU - Huang, Ming Hsi

AU - Chow, Yen Hung

AU - Wang, Jen Ren

AU - Wu, Suh Chin

AU - Liu, Chia Chyi

N1 - Funding Information: This work was supported by the Ministry of Health and Welfare, the Taiwan CDC and the National Health Research Institutes, Taiwan. We also thank the technical services provided by “the i-MANI center of the National Core Facility for Biopharmaceuticals, Ministry of Science and Technology, Taiwan” as well as the technical service from the Instrument Development Center of the National Cheng Kung University. Funding Information: This work was supported by the National Health Research Institutes , Taiwan [Grant Nos. 06A1-IVPP13–014 , 09A1-IVPP13–014 , 11A1-IVPP14–014 , 12A1-IVPP18–014 ]. Funding Information: This work was supported by the National Health Research Institutes, Taiwan [Grant Nos. 06A1-IVPP13–014, 09A1-IVPP13–014, 11A1-IVPP14–014, 12A1-IVPP18–014]. This work was supported by the Ministry of Health and Welfare, the Taiwan CDC and the National Health Research Institutes, Taiwan. We also thank the technical services provided by “the i-MANI center of the National Core Facility for Biopharmaceuticals, Ministry of Science and Technology, Taiwan” as well as the technical service from the Instrument Development Center of the National Cheng Kung University. Publisher Copyright: © 2023 The Author(s)

PY - 2023/5

Y1 - 2023/5

N2 - Coxsackievirus A10 (CVA10) is one of enteroviral pathogens that cause the hand, foot, and mouth disease (HFMD). Since CVA10 was reported to be not easily propagated in the Vero cell culture, a feasible manufacture process for producing formalin-inactivated CVA10 vaccine is urgently needed. Several cell lines that commonly used for viral vaccine production was tested for CVA10 (M2014 strain) culture in this study, and our result showed that CVA10 could be easily propagated in the HEK293A cells. A serum-free HEK293A cell culture system was developed for CVA10 production and the yields have reached over 108 TCID50/mL. The biochemical and immunogenic properties of CVA10 particles obtained from this serum-free HEK293A culture were identical to our previous study. Two major particles of CVA10 were separated by ultracentrifugation, and only the infectious mature particles were capable of inducing CVA10 neutralizing antibody responses in the mouse immunogenicity studies. Additionally, we found that coxsackievirus A6 and enterovirus A71 could also be easily propagated using this serum-free HEK293A cell culture system. Our results provide a solution to overcome the obstacle in the propagation of CVA10 and facilitate the development of multivalent vaccines for prevention of HFMD.

AB - Coxsackievirus A10 (CVA10) is one of enteroviral pathogens that cause the hand, foot, and mouth disease (HFMD). Since CVA10 was reported to be not easily propagated in the Vero cell culture, a feasible manufacture process for producing formalin-inactivated CVA10 vaccine is urgently needed. Several cell lines that commonly used for viral vaccine production was tested for CVA10 (M2014 strain) culture in this study, and our result showed that CVA10 could be easily propagated in the HEK293A cells. A serum-free HEK293A cell culture system was developed for CVA10 production and the yields have reached over 108 TCID50/mL. The biochemical and immunogenic properties of CVA10 particles obtained from this serum-free HEK293A culture were identical to our previous study. Two major particles of CVA10 were separated by ultracentrifugation, and only the infectious mature particles were capable of inducing CVA10 neutralizing antibody responses in the mouse immunogenicity studies. Additionally, we found that coxsackievirus A6 and enterovirus A71 could also be easily propagated using this serum-free HEK293A cell culture system. Our results provide a solution to overcome the obstacle in the propagation of CVA10 and facilitate the development of multivalent vaccines for prevention of HFMD.

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U2 - 10.1016/j.virusres.2023.199101

DO - 10.1016/j.virusres.2023.199101

M3 - Article

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AN - SCOPUS:85150935150

SN - 0168-1702

VL - 329

JO - Virus Research

JF - Virus Research

M1 - 199101

ER -

Propagation and immunological characterization of coxsackievirus A10 in a serum-free HEK293A cell culture system

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