4.7 Article

Rapid production of a H9N2 influenza vaccine from MDCK cells for protecting chicken against influenza virus infection

Journal

APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
Volume 99, Issue 7, Pages 2999-3013

Publisher

SPRINGER
DOI: 10.1007/s00253-015-6406-7

Keywords

H9N2; MDCK; Vaccine; Bioreactor

Funding

  1. National Natural Science Foundation of China [81360312, 81402306]
  2. Guangxi Natural Science Foundation [2014GXNSFBA118158]
  3. Youth Science Foundation of Guangxi Medical University [GXMUYSF201209]
  4. Science and Technology Research Project of the Guangxi Colleges and Universities [YB2014080]
  5. State Key Laboratory of Biocontrol [SKLBC12K11]

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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 5 months, 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.

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