The expression and genetic immunization of chimeric fragment of Hantaan virus M and S segments.
Hemorrhagic fever with renal syndrome (HFRS), which is characterized by severe symptoms and high mortality, is caused by hantavirus. There are still no effective prophylactic vaccines directed to HFRS until now. In this research, we fused expressed G2 fragment of M segment and 0.7kb fragment of S segment. We expect it could be a candidate vaccine. Chimeric gene G2S0.7 was first expressed in prokaryotic expression system pGEX-4T. After inducing expressed fusion proteins, GST-G2S0.7 was induced and its molecular weight was about 100kDa. Meanwhile, the fusion protein kept the activity of its parental proteins. Further, BALB/c mice were vaccinated by the chimeric gene. ELISA, cell microculture neutralization test in vitro were used to detect the humoral immune response in immunized BALB/c mice. Lymphocyte proliferation assay was used to detect the cellular immune response. The results showed that the chimeric gene could simultaneously evoke specific antibody against nucleocapsid protein (NP) and glycoprotein (GP). And the immunized mice of every group elicited neutralizing antibodies with different titers. But the titers were low. Lymphocyte proliferation assay results showed that the stimulation indexes of splenocytes of chimeric gene to NP and GP were significantly higher than that of control. It suggested that the chimeric gene of Hantaan virus containing G2 fragment of M segment and 0.7kb fragment of S segment could directly elicit specific anti-Hantaan virus humoral and cellular immune response in BALB/c mice.
Zhang
Fang-Lin
FL
Department of Microbiology, Fourth Military Medical University, Xi'an, PR China.
Wu
Xing-An
XA
Luo
Wen
W
Bai
Wen-Tao
WT
Liu
Yong
Y
Yan
Yan
Y
Wang
Hai-Tao
HT
Xu
Zhi-Kai
ZK
D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D004926:Escherichia coli / Q000378:metabolism; D005814:Genes, Viral / Q000235:genetics; D006476:Orthohantavirus / Q000276:immunology*; D051379:Mice; D019590:Nucleocapsid Proteins / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000276:immunology*; D014162:Transfection; D014759:Viral Envelope Proteins / Q000276:immunology*; D014765:Viral Vaccines / Q000276:immunology*
D019590:Nucleocapsid Proteins; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; D014765:Viral Vaccines; C090197:glycoprotein G2, Hantavirus