Tweet
Virus Res
. 2026 Apr 2:367:199722.
doi: 10.1016/j.virusres.2026.199722. Online ahead of print.
The influenza B virus candidate vaccine expressing H3 hemagglutinin developed in suspension MDCK cells confers protection against lethal H3N2 avian influenza in BALB/c mice
Yushu Wu 1 , Weiyang Sun 2 , Yeting Xia 3 , Yue Feng 4 , Menglin Zhao 5 , Tiecheng Wang 6 , Xianzhu Xia 7 , Fang Yan 8 , Yuwei Gao 9
Affiliations
Influenza viruses cause annual seasonal epidemics and occasional pandemics. Vaccination is most effective at preventing influenza virus infections. Suspensions of the Madin-Darby canine kidney (MDCKs) cell line are used to manufacture cell-based influenza vaccines. In this study, we tested and optimized a range of culture conditions for a chimeric recombinant influenza virus rA/B-H3 in MDCK suspension cells, revealing that the optimal inoculation conditions were multiplicity of infection of 0.001, cell concentration of infection of 4.0 × 106 cells/mL, addition of 2 μg/mL of L-1-tosylamide-2-phenylethyl chloromethyl ketone, time of harvest of 72 h. The optimal parameter setting in a 5-L bioreactor was 50 % dissolved oxygen and pH 7.2 ± 0.05. The rA/B-H3 has stability genetic after 15 passages in MDCKs. The rA/B-H3 had several immune responsesin mice. Hemagglutination inhibition (HAI) antibodies, micro-neutralizing (MN) antibodies, and IgG antibodies were induced in immunized mice, and the mucosal IgA antibody responses were detected in their lung lavage fluids. The IFN-γ-secretion and IL-4-secretion by the mouse splenocytes were induced after stimulation with the specific H3N2 HA protein. Immunized mice resisted the lethal challenge with a wild-type H3N2 influenza virus. This study demonstrated the reliability of the MDCK suspension cell platform for the production of the chimeric H3N2 candidate vaccine, and the rA/B-H3 candidate vaccine is a potentially safe efficial vaccine.
Keywords: BALB/c mice; Chimeric vaccine; Immunization; MDCK suspension cell; Process optimization.
. 2026 Apr 2:367:199722.
doi: 10.1016/j.virusres.2026.199722. Online ahead of print.
The influenza B virus candidate vaccine expressing H3 hemagglutinin developed in suspension MDCK cells confers protection against lethal H3N2 avian influenza in BALB/c mice
Yushu Wu 1 , Weiyang Sun 2 , Yeting Xia 3 , Yue Feng 4 , Menglin Zhao 5 , Tiecheng Wang 6 , Xianzhu Xia 7 , Fang Yan 8 , Yuwei Gao 9
Affiliations
- PMID: 41935790
- DOI: 10.1016/j.virusres.2026.199722
Influenza viruses cause annual seasonal epidemics and occasional pandemics. Vaccination is most effective at preventing influenza virus infections. Suspensions of the Madin-Darby canine kidney (MDCKs) cell line are used to manufacture cell-based influenza vaccines. In this study, we tested and optimized a range of culture conditions for a chimeric recombinant influenza virus rA/B-H3 in MDCK suspension cells, revealing that the optimal inoculation conditions were multiplicity of infection of 0.001, cell concentration of infection of 4.0 × 106 cells/mL, addition of 2 μg/mL of L-1-tosylamide-2-phenylethyl chloromethyl ketone, time of harvest of 72 h. The optimal parameter setting in a 5-L bioreactor was 50 % dissolved oxygen and pH 7.2 ± 0.05. The rA/B-H3 has stability genetic after 15 passages in MDCKs. The rA/B-H3 had several immune responsesin mice. Hemagglutination inhibition (HAI) antibodies, micro-neutralizing (MN) antibodies, and IgG antibodies were induced in immunized mice, and the mucosal IgA antibody responses were detected in their lung lavage fluids. The IFN-γ-secretion and IL-4-secretion by the mouse splenocytes were induced after stimulation with the specific H3N2 HA protein. Immunized mice resisted the lethal challenge with a wild-type H3N2 influenza virus. This study demonstrated the reliability of the MDCK suspension cell platform for the production of the chimeric H3N2 candidate vaccine, and the rA/B-H3 candidate vaccine is a potentially safe efficial vaccine.
Keywords: BALB/c mice; Chimeric vaccine; Immunization; MDCK suspension cell; Process optimization.