title: DNA mediated immunization with encoding the nucleoprotein gene of porcine transmissible gastroenteritis virus.
abstract: The immune response to a naked plasmid DNA encoding the nucleoprotein (N protein) of porcine transmissible gastroenteritis virus (TGEV) was investigated in this study. A complementary DNA of the entire N gene was amplified by RT-PCR, and inserted into a mammalian expression vector (pcDNA3.1) to construct a recombinant plasmid (pcDNA/N). To evaluate the immunogenicity of the construct, BALB/c mice were intramuscularly immunized with different doses (50, 100 and 200 microg/mouse) of pcDNA/N twice at a 5-week interval. An optimal antibody response was achieved with 100 microg of pcDNA/N. The response lasted at least 11 weeks after primary immunization. By western blotting analysis, the antibodies specifically recognized a 47 kDa protein corresponding to the viral N protein, but they did not reveal neutralizing activity against infectious TGEV in vitro. Immunoglobulin G2a was predominant among these antibodies, which was indicative of Th1 type cell activation in pcDNA/N immunized mice. Moreover, spleen cells from these mice showed stronger immune responses than those from live vaccine or parental vector immunized mice. These results suggest that the construct can elicit both humoral and cell-mediated immune (CMI) responses against TGEV N protein in mice.
authors.affiliation: National Institute of Animal Health, 3-1-1 Kannondai, Tsukuba, 305-0856, Ibaraki, Japan.
authors.forename: C
authors.initials: C
authors.lastname: Liu
authors.forename: T
authors.initials: T
authors.lastname: Kokuho
authors.forename: T
authors.initials: T
authors.lastname: Kubota
authors.forename: S
authors.initials: S
authors.lastname: Watanabe
authors.forename: S
authors.initials: S
authors.lastname: Inumaru
authors.forename: Y
authors.initials: Y
authors.lastname: Yokomizo
authors.forename: T
authors.initials: T
authors.lastname: Onodera
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis / Q000096:biosynthesis; D019556:COS Cells; D018076:DNA, Complementary / Q000276:immunology; D005260:Female; D005761:Gastroenteritis, Transmissible, of Swine / Q000517:prevention & control; D007114:Immunization; D007074:Immunoglobulin G / Q000276:immunology; D008213:Lymphocyte Activation; D051379:Mice; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000276:immunology*; D010957:Plasmids; D013154:Spleen / Q000276:immunology; D013552:Swine; D014162:Transfection; D005760:Transmissible gastroenteritis virus / Q000276:immunology*; D019444:Vaccines, DNA / Q000008:administration & dosage* / Q000276:immunology; D014765:Viral Vaccines / Q000008:administration & dosage* / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D018076:DNA, Complementary; D007074:Immunoglobulin G; D019590:Nucleocapsid Proteins; D019444:Vaccines, DNA; D014765:Viral Vaccines