Hemagglutinin pseudotyped lentiviral particles: characterization of a new method for avian H5N1 influenza sero-diagnosis.
BACKGROUND
Highly pathogenic avian influenza (HPAI) H5N1 has spread globally in birds and infected over 270 humans with an apparently high mortality rate. Serologic studies to determine the extent of asymptomatic H5N1 infection in humans and other mammals and to investigate the immunogenicity of current H5N1 vaccine candidates have been hampered by the biosafety requirements needed for H5N1 micro-neutralization tests.


OBJECTIVE
Development of a serodiagnostic tool for highly pathogenic influenza that reproduces H5N1 biology but can be used with less biohazard.


METHODS
We have generated and evaluated H5 hemagglutinin pseudotyped lentiviral particles encoding the luciferase reporter (H5pp).


RESULTS
H5pp entry into target cells depends on alpha2-3 cell surface sialic acids and requires low pH for membrane fusion. H5pp infectivity is specifically neutralized by sera from patients and animals infected with H5N1 and correlates well with conventional microneutralization test.


CONCLUSIONS
H5pp reproduce H5N1 influenza virus entry into target cells and potentially provides a high-throughput and safe method for sero-epidemiology.
Nefkens
Isabelle
I
HKU-Pasteur Research Center, Hong Kong, SAR China. isabelle.nefkens@gmail.com
Garcia
Jean-Michel
JM
Ling
Chu Shui
CS
Lagarde
Nadège
N
Nicholls
John
J
Tang
Dong Jiang
DJ
Peiris
Malik
M
Buchy
Philippe
P
Altmeyer
Ralf
R
D000818:Animals; D001717:Birds; D004285:Dogs; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics / Q000472:pathogenicity / Q000502:physiology*; D005585:Influenza in Birds / Q000097:blood / Q000175:diagnosis* / Q000821:virology*; D007251:Influenza, Human / Q000097:blood / Q000175:diagnosis* / Q000821:virology*; D016086:Lentivirus / Q000235:genetics / Q000472:pathogenicity / Q000502:physiology*; D008156:Luciferases / Q000235:genetics; D009500:Neutralization Tests / Q000379:methods; D012698:Serologic Tests / Q000379:methods; D014771:Virion / Q000235:genetics / Q000472:pathogenicity / Q000502:physiology*
D019267:Hemagglutinin Glycoproteins, Influenza Virus; C109963:hemagglutinin, avian influenza A virus; D008156:Luciferases