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Development and identification of a new Vero cell-based live attenuated influenza B vaccine by a modified classical reassortment method

tetano

Editor, Senior Moderator
Expert Rev Vaccines. 2017 Jun 5. doi: 10.1080/14760584.2017.1337514. [Epub ahead of print]
[h=1]Development and identification of a new Vero cell-based live attenuated influenza B vaccine by a modified classical reassortment method.[/h] Yang F[SUP]1,[/SUP][SUP]2[/SUP], Ma L[SUP]1[/SUP], Zhou J[SUP]1[/SUP], Wu Y[SUP]1[/SUP], Gao J[SUP]1[/SUP], Song S[SUP]1[/SUP], Geng X[SUP]1[/SUP], Guo Q[SUP]1[/SUP], Li Z[SUP]1[/SUP], Li W[SUP]1[/SUP], Liao G[SUP]1[/SUP].
[h=3]Author information[/h]

[h=3]Abstract[/h] [h=4]OBJECTIVE:[/h] The goal of this study was to generate a new Vero and cold-adapted live attenuated influenza B vaccine and evaluate its safety and immunogenicity with experiments in a guinea pig model.
[h=4]METHODS:[/h] A modified classical reassortment method was used to generate the new vaccine strain. The donor strain was B/Yunnan/2/2005Vca(B), and the parental virus strain was B/Brisbane/60/2008wt. After co-infection in Vero cells, the prepared antibody serum inhibited the donor strain growth, and screening conditions inhibited the candidate virus growth, which induced the growth of the new reassortant virus B/Brisbane/60/2008Vca(B) grow. RT-PCR and sequencing were used to analyze its gene frame composition. The virus proliferation process was observed by immunofluorescence. Through intraperitoneal injection (i.j.) and intranasal injection (n.j.) we evaluated the safety and immunogenicity of the vaccine.
[h=4]RESULTS:[/h] A high-yield of the reassortant virus was produced in Vero cells at 25?C, similar to the donor strains. After sequencing, it was found that B/Brisbane/60/2008Vca(B) Hemagglutinin (HA) and Neuraminidase (NA) gene fragments were from B/Brisbane/60/2008wt, while the other 6 gene fragments were from B/Yunnan/2/2005Vca(B). Additionally, the reassortant virus's intracellular localization and proliferation efficiency under these conditions were observed. Anaphylatoxin testing indicted that the B/Brisbane/60/2008Vca(B) virus was safe for guinea pigs and did not present specific toxicological factors through i.j., and animal weights increased. The n.j. immune pathway experiments showed no significant differences between the treatment and the PBS control group with respect to weight changes (P>0.5). Furthermore, the new strain had a sufficient geometric mean titter (GMT) against B/Brisbane/60/2008wt according to the HI and NI testing. In addition, a secondary injection immunization by n.j. with B/Brisbane/60/2008Vca(B) was required.
[h=4]CONCLUSION:[/h] The new reassortant live attenuated influenza B vaccine was safe and had enough immune stimulating ability.


[h=4]KEYWORDS:[/h] assessment; attenuated vaccine; classical reassortment; influenza B virus

PMID: 28581345 DOI: 10.1080/14760584.2017.1337514
 
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