ABSTRACT
Porcine epidemic diarrhea virus (PEDV) inhibits the host typeⅠinterferon and cellular antiviral response, but its inhibition mechanism is unclear, and the roles of PEDV nonstructural proteins in regulating typeⅠinterferon responses have been seldom studied. To study the effect of nsp1 on typeⅠinterferon response, nsp1 gene was cloned into a eukaryotic expression vector pCAGGS. The expression of nsp1 in transfected cells was determined by Western blot and indirect immunofluorescence assay. The effects of nsp1 on the induction of typeⅠinterferon were evaluated by dual luciferase reporter gene assay, ELISA and VSV bioassay. Western blot and indirect immunofluorescence assay showed that nsp1 was highly expressed in transfected cells and PEDV-infected cells. Dual luciferase reporter gene assay results indicated that nsp1 strongly inhibited the IFN-β promoter activity, and the inhibitory effect was nsp1 dose-dependent. ELISA results showed that nsp1 significantly inhibited the expression of IFN-β in protein level. And VSV replication-inhibition bioassay revealed that nsp1 significantly inhibited typeⅠIFN antiviral activities induced by poly(I:C). Our results implied that nsp1 was a highly conserved protein of PEDV and exhibited antagonistic function on interferon promoter activity. The results have laid a foundation for further understanding the immune evasion mechanism of PEDV and for developing new effective vaccine against PEDV.
ABSTRACT
Since there is no specific symptoms after human have been infected by parvovirus,parvovirus was administered as a potential cancer therapeutics.Parvovirus was employed as an effective treatment in glioblastoma multiforme,Burkitt' s lymphoma,liver cancer,pancreatic carcinoma,breast cancer and colon cancer.Anti-tumor effects of parvovirus were approved to be associated with its oncolytic effects,non-structural protein 1 and immunoregulation.
ABSTRACT
Avian influenza (AI) virus infects both animal and human. Low pathogenic AI virus infections (some H7 and H9 subtypes) have been reported all over the world and pose a potential threat to the poultry industry. Vaccination is the most effective way to prevent virus infection. However, vaccination makes it difficult to differentiate between vaccinated chickens and infected chickens. In order to differentiate vaccinated chickens from naturally infected chickens, we adopted synthetic peptide-based enzyme-linked immunosorbent assay (ELISA) using the peptide sequences from nonstructural protein 1 (NS1) of H9N2. Five synthetic peptides were designed using Protein Variability Sever (http://imed.med.ucm.es/PVS/) and synthesized. NS1-1 ~ NS1-4 peptides failed to detect serum antibodies from both vaccinated and naturally infected chickens. NS1-5 peptide from the C-terminal NS1 protein detected serum antibody from naturally infected chickens but not vaccinated chickens. These results imply that NS1-5 peptide may be a useful tool to differentiate naturally infected chicken from vaccinated chicken as being used in the synthetic peptide-based ELISA.
Subject(s)
Animals , Humans , Antibodies , Chickens , Enzyme-Linked Immunosorbent Assay , Influenza in Birds , Peptides , Poultry , Vaccination , VirusesABSTRACT
Objective To study the correlation between rotavirus nonstructural protein 1(NSP1) and species restriction of the virus and explore the role of NSP1 in the species restriction.Methods NSP1 C-terminal sequences,its whole amino acid sequences from UniProt Database,and its 3' noncoding sequences from GenBank were collected,and intraspecies and interspecies sequence alignment and phylogenetic tree analysis were performed by DNAStar(Lasergene)7.1.Results The diversity of NSP1 between species and rotavirus species restriction hosted a high degree of consistency.NSP1 C-terminal sequence,NSP1 gene 3' noncoding sequence and NSP1 whole sequence shared a high intraspecies identity,which was higher than the identity between species(P