論文

査読有り
2017年2月

Quantitative analysis of the yield of avian H7 influenza virus haemagglutinin protein produced in silkworm pupae with the use of the codon-optimized DNA: A possible oral vaccine

VACCINE
  • Kuniaki Nerome
  • ,
  • Sayaka Matsuda
  • ,
  • Kenichi Maegawa
  • ,
  • Shigeo Sugita
  • ,
  • Kazumichi Kuroda
  • ,
  • Kazunori Kawasaki
  • ,
  • Reiko Nerome

35
5
開始ページ
738
終了ページ
746
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1016/j.vaccine.2016.12.058
出版者・発行元
ELSEVIER SCI LTD

In this study, we aimed to quantitatively compare the increased production of three H7 influenza virus like particle (VLP) haemagglutinin (HA) with the use of a codon-optimized single HA gene in silkworm pupae. Recombinant baculovirus (Korea H7-BmNPV) could produce 0.40 million HA units per pupa, corresponding to 1832}mu g protein. The yield of the HA produced in larva was estimated to be approximately 0.31 million HA units per larva, and there were no significant differences between the three HA proteins. We could establish efficient recovery system of HA production in larvae and pupae with the use of three cycles sonication methods. Next, we compared yields of HA proteins from three different H7 and two H5 recombinant baculoviruses based on the amount of mRNA synthesized in BmN cells, suggesting that mRNA synthesis may be also a useful indicator for the production of HA. Based on HA titres from four recombinants, the yield of HA had a great influence on the codon-optimized effect and the characteristics of the viral HA gene. The recombinant containing codon optimized HA DNA of A/tufted duck/Fukushima/16/2011 (H5N1) did produce more than one million HA units, although another recombinant including of the wild H5N1 strain failed to show HA activity. Electron microscopy revealed the presence of large VLP and small HA particle in the heavy and light fractions. The purified VLPs reacted with the authentic anti-H7 antibodies and the antibodies prepared after immunization with the VLP H7 antigen. Also H5 and H7 VLPs could produce HI antibody in chickens and mice with oral immunization. The antibodies elicited with oral immunization were confirmed in fluorescent antibody analysis and western blotting in Korea H5-BmNPV and H7 HA-BmNPV recombinant infected BmN cells. Taken together, these findings provided important insights into future oral vaccine development. (C) 2017 Elsevier Ltd. All rights reserved.

リンク情報
DOI
https://doi.org/10.1016/j.vaccine.2016.12.058
Web of Science
https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000393257500005&DestApp=WOS_CPL
ID情報
  • DOI : 10.1016/j.vaccine.2016.12.058
  • ISSN : 0264-410X
  • eISSN : 1873-2518
  • Web of Science ID : WOS:000393257500005

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