Giuseppe
Emeritus
J Virol. 2008 Sep 3. [Epub ahead of print]
Growth determinants for H5N1 influenza vaccine seed viruses in MDCK cells.
Murakami S, Horimoto T, Mai LQ, Nidom CA, Chen H, Muramoto Y, Yamada S, Iwasa A, Iwatsuki-Horimoto K, Shimojima M, Iwata A, Kawaoka Y. - Division of Virology, Department of Microbiology and Immunology, and International Research Center for Infectious Diseases, Institute of Medical Science, University of Tokyo, Tokyo, Japan; Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency, Saitama, Japan; National Institute of Hygiene and Epidemiology, Hanoi, Vietnam, Avian Influenza Laboratory, Tropical Disease Centre, Airlangga University, Surabaya, Indonesia; Animal Influenza Laboratory of the Ministry of Agriculture and National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, People's Republic of China; Nippon Institute for Biological Science, Tokyo, Japan; Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, Wisconsin.
H5N1 influenza A viruses are exacting a growing human toll, with more than 240 fatal cases to date.
In the event of an influenza pandemic by these viruses, embryonated chicken eggs, which are the approved substrate for human inactivated vaccine production, will likely be in short supply because chickens will be killed by these viruses or culled to limit the worldwide spread of the infection.
The Madin-Darby canine kidney (MDCK) cell line is a promising alternative candidate substrate because it supports efficient growth of influenza viruses compared to other cell lines.
Here, we addressed the molecular determinants for growth of an H5N1 vaccine seed virus in MDCK cells, revealing the critical responsibility of the Tyr residue at position 360 of PB2, the considerable requirement for functional balance between hemagglutinin (HA) and neuraminidase (NA), and the partial responsibility of the Glu residue at position 55 of NS1.
Based on these findings, we produced a PR8/H5N1 reassortant, optimized for this cell line, that derives all of its genes for its internal proteins from the PR8(UW) strain except for the NS gene, which derives from the PR8(Cambridge) strain, its N1 NA with a long stalk from an early H5N1 strain, and its HA with an avirulent-type cleavage site sequence derived from a circulating H5N1 virus.
Our findings demonstrate the importance and feasibility of a cell-culture based approach to producing seed viruses for H5N1 inactivated vaccines that grow robustly and in a timely, cost-efficient manner as an alternative to egg-based vaccine production.
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PMID: 18768983 [PubMed - as supplied by publisher
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Growth determinants for H5N1 influenza vaccine seed viruses in MDCK cells.
Murakami S, Horimoto T, Mai LQ, Nidom CA, Chen H, Muramoto Y, Yamada S, Iwasa A, Iwatsuki-Horimoto K, Shimojima M, Iwata A, Kawaoka Y. - Division of Virology, Department of Microbiology and Immunology, and International Research Center for Infectious Diseases, Institute of Medical Science, University of Tokyo, Tokyo, Japan; Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency, Saitama, Japan; National Institute of Hygiene and Epidemiology, Hanoi, Vietnam, Avian Influenza Laboratory, Tropical Disease Centre, Airlangga University, Surabaya, Indonesia; Animal Influenza Laboratory of the Ministry of Agriculture and National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, People's Republic of China; Nippon Institute for Biological Science, Tokyo, Japan; Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, Wisconsin.
H5N1 influenza A viruses are exacting a growing human toll, with more than 240 fatal cases to date.
In the event of an influenza pandemic by these viruses, embryonated chicken eggs, which are the approved substrate for human inactivated vaccine production, will likely be in short supply because chickens will be killed by these viruses or culled to limit the worldwide spread of the infection.
The Madin-Darby canine kidney (MDCK) cell line is a promising alternative candidate substrate because it supports efficient growth of influenza viruses compared to other cell lines.
Here, we addressed the molecular determinants for growth of an H5N1 vaccine seed virus in MDCK cells, revealing the critical responsibility of the Tyr residue at position 360 of PB2, the considerable requirement for functional balance between hemagglutinin (HA) and neuraminidase (NA), and the partial responsibility of the Glu residue at position 55 of NS1.
Based on these findings, we produced a PR8/H5N1 reassortant, optimized for this cell line, that derives all of its genes for its internal proteins from the PR8(UW) strain except for the NS gene, which derives from the PR8(Cambridge) strain, its N1 NA with a long stalk from an early H5N1 strain, and its HA with an avirulent-type cleavage site sequence derived from a circulating H5N1 virus.
Our findings demonstrate the importance and feasibility of a cell-culture based approach to producing seed viruses for H5N1 inactivated vaccines that grow robustly and in a timely, cost-efficient manner as an alternative to egg-based vaccine production.
-
PMID: 18768983 [PubMed - as supplied by publisher
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