Expression of African swine fever virus pp62 protein in HEK 293F cells and analy-sis ofinduced immune response
10.16303/j.cnki.1005-4545.2024.12.02
- VernacularTitle:非洲猪瘟病毒pp62蛋白在HEK 293F细胞中的表达及其诱导的免疫应答分析
- Author:
Wenzhu ZHAI
1
;
Ying HUANG
1
;
Chunhao TAO
1
;
Yuheng HE
1
;
Yuanyuan CHU
1
;
Zhen WANG
1
;
Zhongbao PANG
1
;
Hongfei ZHU
1
;
Hong JIA
1
Author Information
1. 中国农业科学院北京畜牧兽医研究所,北京 100193
- Publication Type:Journal Article
- Keywords:
African swine fever virus(ASFV);
eukaryotic expression;
pp62 protein;
immune re-sponse
- From:
Chinese Journal of Veterinary Science
2024;44(12):2514-2520,2555
- CountryChina
- Language:Chinese
-
Abstract:
African swine fever(ASF)is a highly contagious disease caused by the African swine fe-ver virus(ASFV).To evaluate the immunogenicity of the pp62(CP530R)protein of ASFV,the recombinant CP530R protein was expressed in HEK 293F cells transfected with the plasmid pMAL-Fc-CP530R.Six-week-old female C57BL/6J mice were immunized with 10 μg of purified pp62 protein via subcutaneous injection,followed by a booster immunization with the same dosage at 21 days(enhanced).The humoral and cellular immune responses in the mice were then assessed using ELISA,flow cytometry,and ELISpot assays.Western blot analysis confirmed that the pp62 protein was successfully expressed,with a molecular weight of approximately 118.5 kDa.ELISA results indicated that a high level of specific antibodies was detected in the immunized mice,with antibody titers reaching up to 1∶1 638 400 at 7 days after the secondary immunization.The pro-portion of CD8+T lymphocytes in the immunized mice increased compared to the control group(P＜0.05).Results from the Q-PlexTM Mouse Cytokine Screen demonstrated that the secretion levels of IFN-γ,IL-2,IL-4,and IL-10 in serum were significantly upregulated in the immunized mice following secondary immunization(P＜0.001).In summary,these findings indicate that the pp62 protein can significantly stimulate both humoral and cellular immunity in mice,laying the groundwork for further studies on the function of the ASFV pp62 protein and the identification of novel vaccine antigens for ASF.
