Co-expression of Ubiquitin gene and capsid protein gene enhances the potency of DNA immunization of PCV2 in mice

Virology Journal

Xi Li²  Yanjun Zhou³  Guoxin Li³  Guangzhi Tong³  Yuju Yang¹  Yuekun Lang²  Xuesong Li³  Fang Fu² 

[1] Department of Science, Harbin University No. 9 Xuefu Street, Harbin, Heilongjiang, 150086, China;Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences No. 427 Maduan St., Nangang District, Harbin, 150001, China;Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, No. 518 Ziyue Road, Minhang District, Shanghai 200241, China

关键词: Ubiquitin;    Cap;    DNA immunization;    PCV2;   

Others  :  1156620
DOI  :  10.1186/1743-422X-8-264

 received in 2011-03-04, accepted in 2011-05-30,  发布年份 2011

【 摘 要 】

A recombinant plasmid that co-expressed ubiquitin and porcine circovirus type 2 (PCV2) virus capsid protein (Cap), denoted as pc-Ub-Cap, and a plasmid encoding PCV2 virus Cap alone, denoted as pc-Cap, were transfected into 293T cells. Indirect immunofluorescence (IIF) and confocal microscopy were performed to measure the cellular expression of Cap. Three groups of mice were then vaccinated once every three weeks for a total of three doses with pc-Ub-Cap, pc-Cap or the empty vector pCAGGS, followed by challenging all mice intraperitoneally with 0.5 mL 10^6.5 TCID₅₀/mL PCV2. To characterize the protective immune response against PCV2 infection in mice, assays of antibody titer (including different IgG isotypes), flow cytometric analysis (FCM), lymphocyte proliferation, cytokine production and viremia were evaluated. The results showed that pc-Ub-Cap and pc-Cap were efficiently expressed in 293T cells. However, pc-Ub-Cap-vaccinated animals had a significantly higher level of Cap-specific antibody and induced a stronger Th1 type cellular immune response than did pc-Cap-vaccinated animals, suggesting that ubiquitin conjugation improved both the cellular and humoral immune responses. Additionally, viral replication in blood was lower in the pc-Ub-Cap-vaccinated group than in the pc-Cap and empty vector groups, suggesting that the protective immunity induced by pc-Ub-Cap is superior to that induced by pc-Cap.

【 授权许可】

   
2011 Fu et al; licensee BioMed Central Ltd.

【 图 表 】

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