[Characterization of pseudotyped viruses coated with hemagglutinin of H5N1 avian influenza].
OBJECTIVE
To construct pseudovirus bearing H5N1 HA based on a lentivirus vector system. Then we study the biological feature of the pseudovirus. With the newly established viral particles, we performed the serological tests.


METHODS
H5N1 avian influenza virus that isolated from human case was cloned to construct pLP-HA, then pLP-HA co-transfected with lentivirus vector plasmids pLP1, pLP2 and pEmGFP into 293T cells. The supernatant was harvested 48h post-transfection. Concentrated by super centrifuge, the pseudotyped viruses were analyzed by infection test, HA test and micro-neutralization test. At the same time, optimized HA gene and a Vietnam H5N1 HA gene were used to construct pseudotyped virus for comparison.


RESULTS
Pseudotyped virus particles can be observed with electronic microscope. Western-blot revealed that HA glycoprotein can be expressed in the virions. Our neutralization assay by using the pseudoviruses was comparable with the conventional microneutralization assay with wild-type viruses. A high degree of correlation was detected.


CONCLUSIONS
Pseudotyped Viruses coated with HA of H5N1 High Pathogenic Avian Influenza were successfully constructed; it can be used to for the microneutralization assay. The HA gene from different sources affect the efficiency of the packaging of the pseudovirus. But the optimized HA gene can not obviously improve packaging efficiency of the pseudovirus.
Zhang
Yu-song
YS
State Key Laboratory for Viral Genetic Engineering, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
Wang
Da-yan
DY
Gao
Rong-bao
RB
Dong
Jie
J
Li
Zi
Z
Zhang
Ye
Y
Bai
Tian
T
Wang
Min
M
Wen
Le-ying
LY
Shu
Yue-long
YL
D000818:Animals; D002460:Cell Line; D004285:Dogs; D015870:Gene Expression; D005818:Genetic Engineering; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics / Q000502:physiology* / Q000648:ultrastructure; D007251:Influenza, Human / Q000821:virology; D016086:Lentivirus / Q000235:genetics / Q000378:metabolism; D014771:Virion / Q000235:genetics / Q000502:physiology* / Q000648:ultrastructure
D019267:Hemagglutinin Glycoproteins, Influenza Virus