caa a22 4500 605500304 CHVBK 20210128100555.0 cr unu---uuuuu 210128e20150401xx s 000 0 eng 10.1007/s00253-015-6406-7 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6406-7 Rapid production of a H9N2 influenza vaccine from MDCK cells for protecting chicken against influenza virus infection [Elektronische Daten] [Zhenghua Ren, Zhongzheng Lu, Lei Wang, Zeren Huo, Jianhua Cui, Tingting Zheng, Qing Dai, Cuiling Chen, Mengying Qin, Meihua Chen, Rirong Yang] H9N2 subtype avian influenza viruses are widespread in domestic poultry, and vaccination remains the most effective way to protect the chicken population from avian influenza pandemics. Currently, egg-based H9N2 influenza vaccine production has several disadvantages and mammalian MDCK cells are being investigated as candidates for influenza vaccine production. However, little research has been conducted on low pathogenic avian influenza viruses (LPAIV) such as H9N2 replicating in mammalian cells using microcarrier beads in a bioreactor. In this study, we present a systematic analysis of a safe H9N2 influenza vaccine derived from MDCK cells for protecting chickens against influenza virus infection. In 2008, we isolated two novel H9N2 influenza viruses from chickens raised in southern China, and these H9N2 viruses were adapted to MDCK cells. The H9N2 virus was produced in MDCK cells in a scalable bioreactor, purified, inactivated, and investigated for use as a vaccine. The MDCK-derived H9N2 vaccine was able to induce high titers of neutralizing antibodies in chickens of different ages. Histopathological examination, direct immunofluorescence, HI assay, CD4+/CD8+ ratio test, and cytokine evaluation indicated that the MDCK-derived H9N2 vaccine evoked a rapid and effective immune response to protect chickens from influenza infection. High titers of H9N2-specific antibodies were maintained in chickens for 5months, and the MDCK-derived H9N2 vaccine had no effects on chicken growth. The use of MDCK cells in bioreactors for LPAIV vaccine production is an attractive option to prevent outbreaks of LPAIV in poultry. Springer-Verlag Berlin Heidelberg, 2015 H9N2 nationallicence MDCK nationallicence Vaccine nationallicence Bioreactor nationallicence Ren Zhenghua National Engineering Research Center of South China Sea Marine Biotechnology Co., Ltd, Haizhu Technology building, 2429 East Xingang Road, 510330, Guangzhou, People's Republic of China aut Lu Zhongzheng Shenzhen, South China Sea tide Biotechnology Co., Ltd, Minning Garden Office Building Room 905, Cai Tian Road, Futian District, 518000, Shenzhen, Guangdong, People's Republic of China aut Wang Lei Department of Biochemistry, School of Life Sciences, Sun Yat-sen (Zhongshan) University, 135 West Xingang Road, 510275, Guangzhou, People's Republic of China aut Huo Zeren Sun Yat-Sen Memorial Hospital, Sun Yat-Sen (Zhongshan) University, 107 West Yanjiang Road, 510120, Guangzhou, People's Republic of China aut Cui Jianhua Thoracic Surgery of China-Japan Union Hospital, Jilin University, 130033, Changchun, People's Republic of China aut Zheng Tingting Department of Biochemistry, School of Life Sciences, Sun Yat-sen (Zhongshan) University, 135 West Xingang Road, 510275, Guangzhou, People's Republic of China aut Dai Qing Department of Biochemistry, School of Life Sciences, Sun Yat-sen (Zhongshan) University, 135 West Xingang Road, 510275, Guangzhou, People's Republic of China aut Chen Cuiling Department of Biochemistry, School of Life Sciences, Sun Yat-sen (Zhongshan) University, 135 West Xingang Road, 510275, Guangzhou, People's Republic of China aut Qin Mengying Department of Biochemistry, School of Life Sciences, Sun Yat-sen (Zhongshan) University, 135 West Xingang Road, 510275, Guangzhou, People's Republic of China aut Chen Meihua National Engineering Research Center of South China Sea Marine Biotechnology Co., Ltd, Haizhu Technology building, 2429 East Xingang Road, 510330, Guangzhou, People's Republic of China aut Yang Rirong Department of Immunology, School of Preclinical Medicine, Biological Targeting Diagnosis and Therapy Research Center, Guangxi Medical University, 22 Shuangyong Road, 530021, Nanning, Guangxi, People's Republic of China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/7(2015-04-01), 2999-3013 0175-7598 99:7<2999 2015 99 253 https://doi.org/10.1007/s00253-015-6406-7 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6406-7 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Ren Zhenghua National Engineering Research Center of South China Sea Marine Biotechnology Co., Ltd, Haizhu Technology building, 2429 East Xingang Road, 510330, Guangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Lu Zhongzheng Shenzhen, South China Sea tide Biotechnology Co., Ltd, Minning Garden Office Building Room 905, Cai Tian Road, Futian District, 518000, Shenzhen, Guangdong, People's Republic of China aut NATIONALLICENCE 700 1- Wang Lei Department of Biochemistry, School of Life Sciences, Sun Yat-sen (Zhongshan) University, 135 West Xingang Road, 510275, Guangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Huo Zeren Sun Yat-Sen Memorial Hospital, Sun Yat-Sen (Zhongshan) University, 107 West Yanjiang Road, 510120, Guangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Cui Jianhua Thoracic Surgery of China-Japan Union Hospital, Jilin University, 130033, Changchun, People's Republic of China aut NATIONALLICENCE 700 1- Zheng Tingting Department of Biochemistry, School of Life Sciences, Sun Yat-sen (Zhongshan) University, 135 West Xingang Road, 510275, Guangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Dai Qing Department of Biochemistry, School of Life Sciences, Sun Yat-sen (Zhongshan) University, 135 West Xingang Road, 510275, Guangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Chen Cuiling Department of Biochemistry, School of Life Sciences, Sun Yat-sen (Zhongshan) University, 135 West Xingang Road, 510275, Guangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Qin Mengying Department of Biochemistry, School of Life Sciences, Sun Yat-sen (Zhongshan) University, 135 West Xingang Road, 510275, Guangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Chen Meihua National Engineering Research Center of South China Sea Marine Biotechnology Co., Ltd, Haizhu Technology building, 2429 East Xingang Road, 510330, Guangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Yang Rirong Department of Immunology, School of Preclinical Medicine, Biological Targeting Diagnosis and Therapy Research Center, Guangxi Medical University, 22 Shuangyong Road, 530021, Nanning, Guangxi, People's Republic of China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/7(2015-04-01), 2999-3013 0175-7598 99:7<2999 2015 99 253