Semiliki forest virus vector carrying the bovine viral diarrhea virus NS3 (p80) cDNA induced immune responses in mice and expressed BVDV protein in mammalian cells.
Bovine viral diarrhea virus (BVDV) is a primary pathogen responsible for bovine enteric, respiratory and reproductive failure. A genetic region is encoding the p80 (NS3) of BVDV as the most conserved protein among Pestiviruses. BVDV infection in cattle induces NS3 specific lymphocyte proliferation and humoral responses. To generate a DNA vaccine against BVDV, the gene for BVDV-NADL NS3 was cloned into an eukaryotic expression vector of Semiliki Forest virus (pSFV-1). Quadriceps muscles of BALB/c mice were injected with recombinant DNA generated statistically significant cytotoxic T-lymphocyte activity (CTL) and cell mediated immune (CMI) responses against cytopathic and noncytopathic BVDV. Whereas, the BVDV-NS3 did not generate neutralizing antibodies against BVDVin mice. pSFV-1-NS3 DNA was subjected to in vitro transcription into mRNA. The mRNA was transfected into baby hamster kidney cells (BHK-21) and Madin-Darby bovine kidney cells (MDBK). The recombinant cells were used in the detection of DNA antigen responses by immunological assays. This report establishes the ability of BVDV-NS3 DNA inoculation to induce a strong cellular immune responses in mice.
Reddy
J R
JR
JN-International, Inc, Oakland, NE 68045, USA. jnii@trsonline.net
Kwang
J
J
Varthakavi
V
V
Lechtenberg
K F
KF
Minocha
H C
HC
D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis*; D000917:Antibody Formation; D001912:Bovine Virus Diarrhea-Mucosal Disease / Q000517:prevention & control; D002417:Cattle; D002460:Cell Line; D006224:Cricetinae; D018076:DNA, Complementary / Q000276:immunology*; D005822:Genetic Vectors; D051379:Mice; D008807:Mice, Inbred BALB C; D010447:Peptide Hydrolases; D020365:RNA Helicases; D012672:Semliki forest virus; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014162:Transfection; D019444:Vaccines, DNA / Q000276:immunology*; D017361:Viral Nonstructural Proteins / Q000235:genetics*; D014765:Viral Vaccines / Q000276:immunology*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
D000914:Antibodies, Viral; D018076:DNA, Complementary; D010447:Peptide Hydrolases; D020365:RNA Helicases; D019444:Vaccines, DNA; D017361:Viral Nonstructural Proteins; D014765:Viral Vaccines; C086730:p80 protein, bovine viral diarrhea virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases