Generation and characterization of a cold-adapted attenuated live H3N2 subtype influenza virus vaccine candidate.
- Author:
Wen-qi AN
1
;
Peng-hui YANG
;
Yue-qiang DUAN
;
De-yan LUO
;
Chong TANG
;
Wei-hong JIA
;
Li XING
;
Xin-fu SHI
;
Yu-jing ZHANG
;
Xiu-fan LIU
;
Xi-liang WANG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; COS Cells; Cercopithecus aethiops; Dogs; Hemagglutinin Glycoproteins, Influenza Virus; genetics; Influenza A Virus, H3N2 Subtype; immunology; Influenza Vaccines; immunology; Mice; Mice, Inbred BALB C; Neuraminidase; genetics; Plasmids; Reassortant Viruses; immunology; Reverse Transcriptase Polymerase Chain Reaction; Vaccines, Attenuated; immunology; Viral Proteins; genetics
- From: Chinese Medical Journal 2009;122(23):2880-2885
- CountryChina
- Language:English
-
Abstract:
BACKGROUNDH3N2 subtype influenza A viruses have been identified in humans worldwide, raising concerns about their pandemic potential and prompting the development of candidate vaccines to protect humans against this subtype of influenza A virus. The aim of this study was to establish a system for rescuing of a cold-adapted high-yielding H3N2 subtype human influenza virus by reverse genetics.
METHODSIn order to generate better and safer vaccine candidate viruses, a cold-adapted high yielding reassortant H3N2 influenza A virus was genetically constructed by reverse genetics and was designated as rgAA-H3N2. The rgAA-H3N2 virus contained HA and NA genes from an epidemic strain A/Wisconsin/67/2005 (H3N2) in a background of internal genes derived from the master donor viruses (MDV), cold-adapted (ca), temperature sensitive (ts), live attenuated influenza virus strain A/Ann Arbor/6/60 (MDV-A).
RESULTSIn this presentation, the virus HA titer of rgAA-H3N2 in the allantoic fluid from infected embryonated eggs was as high as 1:1024. A fluorescent focus assay (FFU) was performed 24-36 hours post-infection using a specific antibody and bright staining was used for determining the virus titer. The allantoic fluid containing the recovered influenza virus was analyzed in a hemagglutination inhibition (HI) test and the specific inhibition was found.
CONCLUSIONThe results mentioned above demonstrated that cold-adapted, attenuated reassortant H3N2 subtype influenza A virus was successfully generated, which laid a good foundation for the further related research.