[Construction and immunogenic study of recombinant adenovirus containing chimeric gene G2S0.7 and CTL epitopes of Hantaan virus].
OBJECTIVE
To express G2 fragment of M segment and 0.7 kb fragment of S segment and several CTL epitopes of S segment in adenovirus expression system and investigate the immunological properties of hantaan virus chimeric gene.


METHODS
The recombinant adenovirus was constructed and the recombinant adenovirus was obtained after transfecting HEK293 cells. The titer of it was determined and the expressed product was detected by IFA and ELISA. Further, BALB/c mice were vaccinated by the recombinant adenovirus and the immune response was tested by ELISA, microcell-culture neutralizing experiment, T lymphocyte proliferation test (MTT assay) and cell-mediated cytotoxicity assay.


RESULTS
The recombinant adenovirus AG2S0.7CTL1, AG2S0.7CTL2 were constructed successfully and the titer of it was about 10¹⁰-10¹¹ pfu/mL. The expressed protein could be recognized by the hantaan virus NP-specific mAb and glycoprotein G2-specific mAb. The recombinant adenovirus containing CTL epitopes could elicit effectively the cellular immune response aimed to the NP and GP of hantaan virus in BALB/c mice.


CONCLUSIONS
The recombinant adenovirus containing CTL epitopes could induce the higher cellular immune response than the group that not containing CTL epitopes.
Department of Microbiology, Fourth Military Medical University, Xi'an 710032, China.
Gang
G
Hu
Wen-tao
WT
Bai
Xing-an
XA
Wu
Hai-tao
HT
Wang
Zhi-kai
ZK
Xu
Fang-lin
FL
Zhang
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D018984:Epitopes, T-Lymphocyte / Q000235:genetics* / Q000276:immunology; D018063:Hantaan virus / Q000276:immunology*; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology
D000914:Antibodies, Viral; D018984:Epitopes, T-Lymphocyte; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; C090197:glycoprotein G2, Hantavirus