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PubMed1988

Fusion (F) protein gene of Newcastle disease virus: sequence and hydrophobicity comparative analysis between virulent and avirulent strains.

The nucleotide and predicted amino acid sequences have been obtained for the fusion (F) protein gene of the avirulent strain La Sota of Newcastle disease virus (NDV). The F1 N-terminus begins with the tripeptide Leu-Ileu-Gly instead of Phe-X-Gly as usually observed in fusion peptide. It was found that the cleavage-activation domain of the avirulent La Sota strain contained single (but no pairs of) basic residues in the sequence Gly-Arg-Gln-Gly-Arg. Hydrophobicity analysis suggested that the cleavage-activation domain became more hydrophobic and could be less accessible for host-specific protease(s); dibasic residues next to the F1 N-terminus were shown to be important for keeping the cleavage-activation site in exposed positioning, suitable for F protein activation. Comparative sequence analysis of the NDV F proteins revealed a striking homology between lentogenic La Sota and mesogenic Beaudette C strains. Furthermore, 58 variable positions were recorded in the NDV F protein, excluding signal sequence; some of these mutations, in the cysteine-clustered region, were surmised to alter virulence.

Amino Acid Sequence↗
PubMed1986

[Demonstration of the M-protein of the Newcastle disease virus by current immunochemical methods].

The immunosorbent micromethod with the use of nitrocellulose paper was employed for the first time to demonstrate subviral antigens from animal origin, such as the M-protein of the Newcastle Disease virus. The method proved to be highly effective. It was useful in discovering amounts as high as 10-20 ng of the antigen, while with the use of the enzyme-linked immunosorbent assay (ELISA) amounts of about 200 ng could be evaluated. The immunosorbent micromethod performed on nitrocellulose paper was readily applicable as compared to all previously employed highly sensitive methods, and could be used successfully in immunochemical investigations.

Animals↗
PubMed1993

Analysis of pathotype-specific structural features and cleavage activation of Newcastle disease virus membrane glycoproteins using antipeptide antibodies.

Peptides were synthesized, that correspond to cleaved and trimmed carboxyl termini of the F2 polypeptide regions of fusion (F) protein precursors (F0 proteins) in four different strains of Newcastle disease virus (NDV). These peptides differed only within the four carboxyl-terminal residues and represent F2 polypeptides of virulent (AV), low-virulence (EG) and avirulent (V4 and WA) pathotypes of NDV. Polyclonal rabbit antisera against each peptide reacted with their corresponding monomeric F2 polypeptides and F protein oligomers as analysed by immunoblotting of egg-propagated virions. Bidirectional cross-reactivity was observed between V4 and EG antisera and F2 polypeptides which differ only by a single variation of lysine and arginine at position 3 from their carboxyl termini. The other two antisera (AV and WA) were specific for their corresponding F2 polypeptides. All of these antisera were shown to react in a strain-specific manner with intact egg-propagated virions in an ELISA. A previously described antiserum, designed to target the haemagglutinin-neuraminidase (HN) protein precursor (HN0 protein) of avirulent strains of NDV, has been shown to be specific for residual HN0 protein of avirulent virions propagated in embryonated chicken eggs. Whereas the antiserum targeted at the carboxyl terminus of the V4 F2 polypeptide did not react with F0 proteins of cell culture-propagated strains in immunoblotting, antipeptide antibodies targeted at another region of the F2 polypeptide and a segment of the F1 polypeptide did react with the F0 protein from infected cells. These data are consistent with inclusion of the terminal carboxylate of the F2 polypeptides in the recognition determinants of the antibodies targeted at the carboxyl terminus of the V4 F2 polypeptide. The antisera described herein are ideally suited to rapid immunochemical pathotyping of NDV isolates and immunochemical characterization of the sites of intracellular cleavage activation of F0 and HN0 proteins and may be useful for defining interactions involved in F protein folding.

Amino Acid Sequence↗
PubMed1981

Noncytopathic mutants of Newcastle disease virus are defective in virus-specific RNA synthesis.

We have studied virus-specific RNA synthesis in cells infected by six noncytopathic (nc) mutants of the Australia-Victoria wild-type strain (AV-WT) of Newcastle disease virus (NDV) (19). The rates of NDV-specific RNA synthesis in mutant infection were three to sevenfold lower than those observed in wild-type infection. Velocity sedimentation of this NDV-specific RNA revealed that the lower rates of synthesis in mutant infection correlated with reduced accumulation of 18S and 35S mRNA. Electrophoresis in polyacrylamide-urea gels showed that accumulation of all of the 18S mRNA species was reduced and no new species could be detected. Primary transcription appeared unaltered in mutant infection. Cells infected with two naturally occurring avirulent strains of NDV also showed less accumulation of 18S mRNA. Electrophoresis of this RNA resulted in patterns which differed from those obtained with RNA from either AV-WT or nc mutant infection. Complementation for RNA accumulation between the nc mutants and RNA- temperature-sensitive mutants of AV-WT (32) suggested a common defect in the nc mutants. Analysis of plaque-forming revertants of five of the nc mutants revealed that viral RNA synthetic capacity, cell killing, and plaque-forming ability correlated absolutely. These results suggest that viral RNA synthesis and cytopathogenicity may be causally related. In addition, several of the plaque-forming (and cell-killing) revertants were found to be unable to induce fusion from within in infected cell cultures. This result, coupled with the finding that several of the nc mutants are capable of wild-type levels of fusion from within, suggests that the ability to cause such fusion does not correlate with the ability to kill cells.

Animals↗
PubMed1977

[Sensitivity testing of chick embryo subcultures to the Newcastle disease virus].

A total of five passages of subcultures were produced from chick embryos with the use of a medium enriched with a 10 per cent solution the previous passages of the cells. The form, number, and size of the plaques obtained at the cultivation of the Newcastle disease virus in the cells of the various passages of the subcultures did not differ. The virus production per cell did not also change substantially with the increase in the number of subculture passages.

Animals↗
PubMed2005

[Generation of newcastle disease virus strain ZJI isolated from an outbreak in the goose using reverse genetics technique].

The full-length cDNA clone, NDV3GM122, and the three helperplasmids pCI-NP, pCI-P and pCI-L of Newcastle disease virus strain ZJI isolated from an outbreak in the goose were cotransfected into BSR-T7/5 cell expressing T7 RNA polymerase. Meanwhile, the full-length cDNA clone NDV3GM122 and the three helperplasmids, pCIneoNP, pCIneoP and pCIneoL which were derived from NDV strain La Sota, were also cotransfected into the cell, respectively. Indiect immunofluorescence assay (IFA) was performed 48 to 96 hours post-transfection using NDV HN-specific monoclonal anbtibody (McAb) 6B1 and bright stainings were found in the transfectants, indicating that the full-length clone was functional and the HN protein was expressed. The transfected cell and the supernatant were mixed well and thereafter the mixture was inoculated into specific pathogen free (SPF) chicken eggs. The allanotoic fluid of the injected eggs gave a positive hemagglutinin( HA) titer ranging from 16 to 32 in the secondary passage and increased to 128 in the third passage, which was same to the level of parent wild-type virus. The allantoic fluid containing the recovered NDV was analyzed in hemagglutination inhibition( HI) test by using McAb 6B1 and the specific inhibition was found. The typical morphology of the produced NDV was detected in the electronic microscope. The results mentioned above demonstrated that infectious NDV of strain ZJI was successfully generated, which laid good foundation for the further related research.

Animals↗
PubMedDate not supplied

Detection of antibodies to Newcastle disease virus in ostriches (Struthio camelus) by an indirect ELISA.

A two-graph receiver operating characteristic analysis, performed on the hemagglutination-inhibition (HI) and enzyme-linked immunosorbent assay (ELISA) test results of a Newcastle disease virus (NDV)-positive and NDV-negative control group of ostrich sera, proved that the ELISA was superior to the HI in both sensitivity and specificity. Comparison of results of the two assays performed on a panel of simulated positive sera ranging from very weak to very strong showed that the ELISA was at least 10 times more sensitive than the HI in detecting low levels of ostrich antibodies to NDV. The ELISA also has the advantage of using untreated serum in a single dilution as opposed to the HI test, which uses pretreated serum in titration.

Animals↗
PubMedDate not supplied

The pathogenesis of velogenic Newcastle disease virus infection of chickens of different ages and different levels of immunity.

Chickens of 7 weeks or 20 weeks of age were divided into three groups according to their antibody status (high, low, absent) and were infected with a velogenic viscerotropic Newcastle disease virus. To follow patterns of viral replication, birds were necropsied at regular intervals up to 22 days and organs were sampled from each bird. In non-immune birds, virus could be isolated from all organs examined. In birds with antibody, virus was most frequently isolated from the proventriculus, cecal tonsil, bursa, and brain. However, because no one organ could be recommended for all situations, all four should be sampled for field diagnosis. In immune birds, although clinical signs were either mild or absent, widespread virus replication occurred up to 19 days post-challenge.

Animals↗
PubMed1984

Neutralization of Newcastle disease virus by monoclonal antibodies to the hemagglutinin-neuraminidase glycoprotein: requirement for antibodies to four sites for complete neutralization.

The neutralizing characteristics of monoclonal antibodies directed to four antigenic sites on the hemagglutinin-neuraminidase glycoprotein of Newcastle disease virus were determined. Neutralization by each antibody resulted in a persistent fraction of nonneutralized virus which varied from 1 to 17% depending on the hemagglutinin-neuraminidase site recognized, but not on the antibody. The addition of antibodies to all four sites on the hemagglutinin-neuraminidase glycoprotein was required to give a level of neutralization comparable with that obtained with polyclonal mouse antiserum. The high persistent fractions were not due to viral aggregates, a high level of variants in the virus stock, the use of insufficient antibody, low antibody avidity, or an effect peculiar to the use of the chicken cells as host. The addition of rabbit anti-mouse immunoglobulin to the persistent fraction left by any of the antibodies resulted in a further reduction in infectivity, often by as much as two logs. Thus, some viral particles are capable of binding antibody while retaining their infectivity. The implications of these findings to the mechanism of neutralization are discussed.

Allantois↗
PubMed1994

Genetic analysis of antibody responses of turkeys to Newcastle disease virus and Pasteurella multocida vaccines.

Heritability (h2) of 16-wk BW and primary and secondary antibody responses and genetic and phenotypic correlations among these traits were estimated for 931 male and female turkeys vaccinated with Newcastle disease virus (NDV) and Pasteurella multocida. Turkeys from a line selected for 22 or 23 generations for increased 16-wk BW were vaccinated at 6 and 12 wk of age with blood samples collected 3 wk postvaccination. Antibody titers were determined using an ELISA method and transformed to log(e) for analysis. Heritability estimates for primary and secondary antibody responses to NDV were .380 +/- .070 (SE) and .296 +/- .063, respectively. For primary and secondary antibody responses to P. multocida, h2 estimates were .458 +/- .075 and .333 +/- .066, respectively. Heritability estimate for 16-wk BW was .404 +/- .071. The genetic correlation between primary and secondary antibody responses to NDV was .491 +/- .150. There was no genetic correlation between primary and secondary antibody responses to P. multocida. Although the genetic correlation between primary antibody responses to NDV and P. multocida was .292 +/- .159, the genetic correlation between secondary responses to the two antigens did not differ from zero. There were no genetic correlations between antibody responses and 16-wk BW. Similar results were observed for phenotypic correlations. Based on heritability and genetic correlation estimates, it would be possible to improve antibody responses to either NDV or P. multocida singularly; however, to improve antibody responses to both antigens, selection would have to be applied for each antigen.

Animals↗
PubMed1999

Comparison of pathology-based techniques for detection of viscerotropic velogenic Newcastle disease virus in chickens.

Two pathology-based techniques, immunohistochemistry and riboprobe in-situ hybridization, were applied to formalin-fixed, paraffin wax-embedded tissues from chickens infected with three different isolates of velogenic viscerotropic Newcastle disease virus (VVNDV). With the immunohistochemical method, viral protein was consistently detectable in the spleen and caecum at the terminal phase of the infection. With in-situ hybridization, viral nucleic acid was consistently detected in the eyelid, spleen and caecum in both the acute and terminal phases. Hybridization with anti-sense probe to detect viral mRNA was often more intense than hybridization with sense probe to detect viral genomic RNA.

Animals↗
PubMed2005

Molecular characterization and phylogenetic analysis of Newcastle disease virus isolates from healthy wild birds.

Wild waterfowl is considered a natural reservoir of potentially infectious agents and a source of pathogenic viruses like avian paramyxoviruses type 1 (APMV 1). In 1997, commercial poultry in Argentina had reached the status of being free from virulent Newcastle disease virus (NDV) infections. Vaccination and biosecurity measures are actively performed to maintain this preferential sanitary condition. However, the risk of reintroduction of pathogenic viruses is always present. In this context, we conducted a study to describe the status of wild healthy birds in a geographic region relevant for the poultry industry. The presence of anti-NDV antibodies was determined in different species in all areas sampled suggesting previous contact with NDV. Seven ND viruses were isolated and characterized as apathogenic strains by biological and molecular methods. The phylogenetic analysis revealed that the majority of the Argentinian isolates form a subgroup related to viruses of genotype II. The results presented here highlight the importance of maintaining strict biosecurity measures and vaccination programs in poultry industries in order to preserve the virulent NDV-free status for commercial flocks in the country.

Amino Acid Sequence↗
PubMed1972

Viral ribonuclei acid synthesis by Newcastle disease virus mutants isolated from persistently infected L cells: effect of interferon.

The synthesis of different viral ribonucleic acid (RNA) species was studied in chick embryo (CE) and mouse L-cell cultures infected with the Herts strain of Newcastle disease virus (NDV(o)) and a mutant isolated from persistently infected L cells (NDV(pi)). In CE cell cultures, both viruses synthesized significant amounts of 54, 36, and 18S RNA. However, in L cells, synthesis of 54S virion RNA was markedly reduced. From these results, it seems likely that the low yield of infective virus in L cells is due to a deficient synthesis of 54S RNA in this host. On this basis, however, it is apparent that the "covert" replication of NDV(o) in L cells is due to factors other than viral RNA synthesis. When low concentrations of interferon were used to pretreat CE cells, a differential effect on the synthesis of various RNA species was observed. The 18S RNA of NDV(o) was more sensitive to interferon action than the 36 and the 54S RNA species. In contrast, the 18S RNA of NDV(pi) was less sensitive than the 36S and the 54S RNA. The inhibition of 54S RNA synthesis correlated with the reduction of viral yield and explained the greater sensitivity of NDV(pi) to interferon.

Animals↗
PubMed1999

[The nucleotide sequences of the HN gene and F gene fragment of Newcastle disease virus strains BOR74 and BOR82].

The complete nucleotide sequence of HN gene, the region of F gene, and intergene regions (M-F, F-HN, and HN-L) of the BOR74 and BOR82 strains of Newcastle disease virus have been determined. Based on the nucleotide and amino acid sequences, the speeds of the nucleic and amino acid changes were calculated (approximately 10(-3) nucleotides or amino acids/year). The BOR strains were grouped phylogenetically with the asymptomatic strains. These strains and the BOR strains have the same motif of the cleavage site (112GKQGR116-L117), but the HN protein of BOR strains has the 572 amino acids which differ the BOR strains from all other strains (571, 577, and 616 amino acids).

Base Sequence↗
PubMed1979

Interferon induction with Newcastle disease virus in FS-4 cells: effect of 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB).

DRB is an inhibitor of heterogeneous nuclear RNA (hnRNA) and messenger RNA (MRNA) synthesis. The effect of DRB on interferon production stimulated by Newcastle disease virus (NDV) in the human FS-4 cells was studied. Interferon production in cells primed by treatment with interferon was markedly enhanced (superinduced) in the presence of DRB. This superinduction was essentially due to an inhibition of the rapid decline (shutoff) of interferon production observed in primed cells not treated with DRB. Continuous presence of DRB was required for maximal superinduction. In this and other respects the interferon response induced by NDV in primed cells resembled poly(I). poly(C)-induced interferon production. In contrast interferon production in cells not primed with interferon was virtually abolished by DRB treatment. Since neither virus specific RNA synthesis nor virus replication were significantly affected by DRB, the inhibition of interferon production is likely to result from the inhibitory action of DRB on a cellular, rather than viral, function. Apparently some differences exist in the synthesis or processing of the mRNAs for interferons in primed and unprimed cells and these determine the different sensitivities of these two responses to DRB.

Cell Line↗