Construction and immunogenicity study of 293T cell line stably expressing rabies virus glycoprotein
10.16303/j.cnki.1005-4545.2024.11.12
- VernacularTitle:稳定表达狂犬病病毒糖蛋白293T细胞系的构建及免疫原性研究
- Author:
Yu WANG
1
,
2
;
Shoufeng ZHANG
;
Tingting YU
;
Lijuan MI
;
Naiquan YAO
;
Faming MIAO
;
Rongliang HU
Author Information
1. 吉林农业大学动物医学院,吉林长春 130118
2. 中国农业科学院长春兽医研究所,吉林 长春 130122
- Keywords:
rabies;
rabies virus glycoprotein;
lentivirus;
cell lines;
immunological experiment;
subunit vaccine
- From:
Chinese Journal of Veterinary Science
2024;44(11):2393-2399
- CountryChina
- Language:Chinese
-
Abstract:
In order to obtain a cell line which high expressed of rabies virus glycoprotein the produc-tion of rabies subunit vaccine,the RABV glycoprotein gene sequence was amplified by RT-PCR and cloned into the lentiviral expression vector.The recombinant plasmid pLV-RVG,envelope plasmid pMD2.0G and packaging plasmid psPAX2 were co-transfected into HEK-293T cells to harvest lentivirus and infect new HEK-293T cells.The 293T-RVG cell line was obtained by puro-mycin pressure screening.Indirect immunofluorescence assay(IFA)and Western blot showed that the 67 kD RABV-G protein was highly expressed in the cells.The neutralizing antibody titer reached 2.19 IU/mL within 14 days after immunization,which was higher than the internationally recognized protection threshold(0.5 IU/mL).This study showed that the cell line 293T-RVG with stable and high expression of rabies virus glycoprotein was successfully constructed.The cell line could protect mice against rabies virus challenge and could be used as a subunit vaccine with poten-tial for large-scale production and application.
