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Co-expression of ubiquitin gene and capsid protein gene enhances the potency of DNA immunization of PCV2 in mice.
- Source :
-
Virology journal [Virol J] 2011 May 30; Vol. 8, pp. 264. Date of Electronic Publication: 2011 May 30. - Publication Year :
- 2011
-
Abstract
- 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 10⁶·⁵ 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.
- Subjects :
- Animals
Antibodies, Viral blood
Capsid Proteins genetics
Cell Line
Cell Proliferation
Circovirus genetics
Cytokines metabolism
Female
Flow Cytometry
Gene Expression
Humans
Immunization, Secondary methods
Immunoglobulin G blood
Lymphocyte Subsets immunology
Mice
Mice, Inbred BALB C
Microscopy, Confocal
Plasmids administration & dosage
Ubiquitin genetics
Vaccination methods
Vaccines, DNA administration & dosage
Vaccines, DNA genetics
Viremia prevention & control
Capsid Proteins biosynthesis
Circovirus immunology
Ubiquitin biosynthesis
Vaccines, DNA immunology
Subjects
Details
- Language :
- English
- ISSN :
- 1743-422X
- Volume :
- 8
- Database :
- MEDLINE
- Journal :
- Virology journal
- Publication Type :
- Academic Journal
- Accession number :
- 21624113
- Full Text :
- https://doi.org/10.1186/1743-422X-8-264