Comparing the antibody responses against recombinant hemagglutinin proteins of avian influenza A (H5N1) virus expressed in insect cells and bacteria

The hemagglutinin (HA) of influenza A virus plays an essential role in mediating the entry of the virus into host cells. Here, recombinant full-length HA5 protein from a H5N1 isolate (A/chicken/hatay/2004(H5N1)) was expressed and purified from the baculovirus-insect cell system. As expected, full-le...

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Bibliographic Details
Published in:Journal of medical virology 2008-11, Vol.80 (11), p.1972-1983
Main Authors: Shen, Shuo, Mahadevappa, Geetha, Oh, Hsueh-Ling Janice, Wee, Boon Yu, Choi, Yook-Wah, Hwang, Le-Ann, Lim, Seng Gee, Hong, Wanjin, Lal, Sunil K, Tan, Yee-Joo
Format: Article
Language:English
Subjects:
Animals
Antibodies, Viral - blood
Bacteria
Baculoviridae - genetics
baculovirus
Biological and medical sciences
Cell Line
Fundamental and applied biological sciences. Psychology
Genetic Vectors
HA5 pseudotyped lentiviral particles
Hemagglutinin Glycoproteins, Influenza Virus - genetics
Hemagglutinin Glycoproteins, Influenza Virus - immunology
Human viral diseases
Infectious diseases
Influenza A virus
Influenza A Virus, H5N1 Subtype - genetics
Influenza A Virus, H5N1 Subtype - immunology
Influenza Vaccines - immunology
Medical sciences
membrane fusion
Microbiology
Miscellaneous
Neutralization Tests
neutralizing antibodies
Recombinant Proteins - genetics
Recombinant Proteins - immunology
Spodoptera
Viral diseases
Virology
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Summary:The hemagglutinin (HA) of influenza A virus plays an essential role in mediating the entry of the virus into host cells. Here, recombinant full-length HA5 protein from a H5N1 isolate (A/chicken/hatay/2004(H5N1)) was expressed and purified from the baculovirus-insect cell system. As expected, full-length HA5 elicits strong neutralizing antibodies, as evaluated in micro-neutralization tests using HA5 pseudotyped lentiviral particles. In addition, two fragments of HA5 were expressed in bacteria and the N-terminal fragment, covering the ectodomain before the HA1/HA2 polybasic cleavage site, was found to elicit neutralizing antibodies. But the C-terminal fragment, which covers the remaining portion of the ectodomain, did not. Neutralizing titer of the anti-serum against the N-terminal fragment is only four times lower than the anti-serum against the full-length HA5 protein. Using a novel membrane fusion assay, the abilities of these antibodies to block membrane fusion were found to correlate well with the neutralization activities. J. Med. Virol. 80:1972-1983, 2008.
ISSN:0146-6615
1096-9071
DOI:10.1002/jmv.21298