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PubMed1971

Enhancement of bacterial infections in mice by newcastle disease virus.

This report describes an attempt to define the factors which incite secondary bacterial pneumonias. Groups of mice were given bacteria intraperitoneally and, at various intervals, Newcastle disease virus intravenously. There was an increase in the number of deaths and in the rates of death in these groups, when compared with a control group which was given only bacteria. These results were obtained with Streptococcus pneumoniae (Diplococcus pneumoniae), Pseudomonas aeruginosa, and Salmonella enteritidis ser. typhimurium. Newcastle disease virus increased the mortality rate of mice with bacterial infections when the two agents were given within 24 hr.

Journal Article↗
PubMed2002

Nucleotide sequence analysis of the Newcastle disease virus nucleocapsid protein gene and phylogenetic relationships among the Paramyxoviridae.

The nucleocapsid (N) protein genes from 24 Newcastle disease virus (NDV) isolates representing various pathotypes with different geographical and chronological origins were cloned and sequenced. The N-terminal region of the N protein to residue 401 was highly conserved among isolates with several conservative substitutions occurring that correlated with phylogenetic relationships. Variability of the N protein was detected in the C-terminal portion similar to what has been reported for other members of the Paramyxovirinae. Amino acids previously identified as invariant or highly conserved in N proteins of other paramyxoviruses were also present in the NDV protein. Phylogenetic analysis of N gene coding sequences among NDV isolates again demonstrated the existence of two major groups. One clade contained viruses that included vaccine and virulent strains isolated in the USA prior to 1970 while a second clade included vaccine and virulent viruses isolated worldwide. Comparison of N protein amino acid sequences among members of the Paramyxoviridae resulted in NDV and avian paramyxovirus 6 separating as a cluster distinct from the Rubulavirus genus. This provides further support for avian paramyxoviruses being considered for their own genus among the Paramyxovirinae.

Amino Acid Sequence↗
PubMed2002

Newcastle disease virus (NDV) marker vaccine: an immunodominant epitope on the nucleoprotein gene of NDV can be deleted or replaced by a foreign epitope.

The nucleoprotein (NP) of Newcastle disease virus (NDV) functions primarily to encapsidate the virus genome for the purpose of RNA transcription, replication, and packaging. This conserved multifunctional protein is also efficient in inducing NDV-specific antibody in chickens. Here, we localized a conserved B-cell immunodominant epitope (IDE) spanning residues 447 to 455 and successfully generated a recombinant NDV lacking the IDE by reverse genetics. Despite deletion of NP residues 443 to 460 encompassing the NP-IDE, the mutant NDV propagated in embryonated specific-pathogen-free chicken eggs to a level comparable to that of the parent virus. In addition, a B-cell epitope of the S2 glycoprotein of murine hepatitis virus (MHV) was inserted in-frame to replace the NP-IDE. Recombinant viruses properly expressing the introduced MHV epitope were successfully generated, demonstrating that the NP-IDE not only is dispensable for virus replication but also can be replaced by foreign sequences. Chickens immunized with the hybrid recombinants produced specific antibodies against the S2 glycoprotein of MHV and completely lacked antibodies directed against the NP-IDE. These marked-NDV recombinants, in conjunction with a diagnostic test, enable serological differentiation of vaccinated animals from infected animals and may be useful tools in ND eradication programs. The identification of a mutation-permissive region on the NP gene allows a rational approach to the insertion of protective epitopes and may be relevant for the design of NDV-based cross-protective marker vaccines.

Amino Acid Sequence↗
PubMed1996

Growth of Newcastle disease virus in chicken macrophages.

Chicken macrophages were isolated from chicken peripheral blood, cultured in vitro, and infected with Newcastle disease virus (NDV) after reaching semiconfluence. Infected macrophages supported the growth and replication of NDV. Virus-infected macrophages exhibited features of apoptosis as determined by agarose gel electrophoresis, thymidine release assays and flow cytometry. Electron microscopical examination also showed the appearance of intact virus particles, fragmented chromatin and apoptotic bodies. This evidence demonstrates that NDV infection of chicken macrophages causes chicken macrophages to undergo apoptosis.

Animals↗
PubMed2000

Performance of an RT-nested PCR ELISA for detection of Newcastle disease virus.

A sensitive and specific RT-nested PCR coupled with an ELISA detection system for detecting Newcastle disease virus is described. Two nested pairs of primer which were highly specific to all the three different pathotypes of NDV were designed from the consensus fusion gene sequence. No cross-reactions with other avian infectious agents such as infectious bronchitis virus, infectious bursal disease virus, influenza virus, and fowl pox virus were observed. Based on agarose electrophoresis detection, the RT-nested PCR was about 100 times more sensitive compared to that of a non-nested RT-PCR. To facilitate the detection of the PCR product, an ELISA detection method was then developed to detect the amplified PCR products and it was shown to be ten times more sensitive than gel electrophoresis. The efficacy of the nested PCR-ELISA was also compared with the conventional NDV detection method (HA test) and non-nested RT-PCR by testing against a total of 35 tissue specimens collected from ND-symptomatic chickens. The RT-nested PCR ELISA found NDV positive in 21 (60%) tissue specimens, while only eight (22.9%) and two (5.7%) out of 35 tissue specimens were tested NDV positive by both the non-nested RT-PCR and conventional HA test, respectively. Due to its high sensitivity for the detection of NDV from tissue specimens, this PCR-ELISA based diagnostic test may be useful for screening large number of samples.

Animals↗
PubMed1982

Maturation of the envelope glycoproteins of Newcastle disease virus on cellular membranes.

Based on subcellular fractionation data, the following maturation pathways were proposed for the Newcastle disease virus glycoproteins. During or shortly after synthesis in rough endoplasmic reticulum, hemagglutinin-neuraminidase (HN) and fusion (F0) glycoproteins underwent dolichol pyrophosphate-mediated glycosylation, and HN assumed a partially trypsin-resistant conformation. HN began to associate into disulfide-linked dimers in rough endoplasmic reticulum, and at least one of its oligosaccharide side chains was processed to a complex form en route to the cell surface. During migration in intracellular membranes, F0 was proteolytically cleaved to F1.2. Neither HN nor F1,2 required oligosaccharide side chains for migration to plasma membranes, and cleavage of F0 also occurred without glycosylation. Virion- and plasma membrane-associated HN contained both complex and high-mannose oligosaccharide chains on the same molecule, and F1,2 contained at least high-mannose forms. Several of the properties of HN were notable for a viral glycoprotein. The oligosaccharide side chains of HN were modified very slowly in chick cells, whereas those of the G glycoprotein of vesicular stomatitis virus were rapidly processed to a complex form. Therefore, their different rates of migration and carbohydrate processing were intrinsic properties of these glycoproteins. Consistent with its slow maturation, the HN glycopolypeptide accumulated to high levels in intracellular membranes as well as in plasma membranes. Intracellular HN contained immature oligosaccharide side chains, suggesting that it accumulated in the pre-Golgi/Golgi segment of the maturation pathway. The major site of accumulation of mature HN with neuraminidase activity was the plasma membrane.

Animals↗
PubMed1992

Fusion mutants of Newcastle disease virus selected with monoclonal antibodies to the hemagglutinin-neuraminidase.

The Australia-Victoria (AV) isolate of Newcastle disease virus (NDV) induces fusion from within but not fusion from without. L1, a neuraminidase (NA)-deficient virus derived from AV, has the opposite fusion phenotype from the wild-type virus. It fails to induce the former mode of fusion, but has gained a limited ability to promote the latter. Monoclonal antibodies to antigenic site 23 on the hemagglutinin-neuraminidase (HN) glycoprotein have previously been shown to select variants of the AV isolate that have altered NA activity or receptor-binding affinity. By using an antibody to this site, variants of L1 have been selected. Three of the variants have gained an increased affinity for sialic acid-containing receptors, as evidenced by the resistance of their hemagglutinating activity to the presence of reduced amounts of sialic acid on the surface of chicken erythrocytes. All four variants still have very low levels of NA activity, comparable to that of the parent virus, L1. The alteration in receptor-binding affinity results in a decreased potential for elution from cellular receptors and correlates with an increased ability to promote both modes of fusion. A single amino acid substitution in the HN protein of each variant, responsible for its escape from neutralization, has been identified. These studies identify two HN residues, 193 and 203, at which monoclonal antibody-selected substitution influences the receptor recognition properties of NDV and may influence its ability to promote syncytium formation.

Amino Acid Sequence↗
PubMed1991

Identification of endoprotease activity in the trans Golgi membranes of rat liver cells that specifically processes in vitro the fusion glycoprotein precursor of virulent Newcastle disease virus.

A ubiquitous host endoprotease(s) responsible for activation of the fusion glycoprotein precursor (F0) of virulent Newcastle disease virus (NDV) is an important determinant for its spreading and organ tropism in the host. To characterize the virus-activating protease (VAP), we isolated endoprotease activity from the trans Golgi membranes of rat liver cells by using F0-containing NDV particles grown in a lymphoid cell line NALM6 as substrate. The enzyme cleaved in vitro only the F0 protein of virulent NDV but not that of an avirulent strain, suggesting that it specifically recognizes pairs of basic residues at the cleavage site. Furthermore, the enzyme was found to be membrane-bound, calcium ion-dependent, and active over a broad pH range, from 6 to 8. The inhibitor spectrum of the protease together with the enzyme properties described above indicates that it is a KEX2-like enzyme. Experiments using monensin, A23187, and chloroquine indicate that the F0 cleavage of virulent NDV occurs normally in rat primary hepatocytes at or before the trans Golgi and is a calcium-dependent process. The correspondence between the characteristics of the cleavage in rat hepatocytes and those of the rat protease in vitro indicates that the endoprotease is a strong candidate for the VAP that determines the pantropic nature of virulent NDV.

Animals↗
PubMed1986

Nucleotide sequence of the gene encoding the matrix protein of Newcastle disease virus.

The nucleotide sequence of the gene encoding the matrix (M) protein of the Beaudette C strain of Newcastle disease virus (NDV) has been determined from overlapping cDNA clones. Control sequences typical of paramyxovirus mRNA start and polyadenylation signals have been identified. Assuming that the M gene starts and finishes at these sequences, the M gene is 1241 nucleotides long and encodes one long open reading frame of 364 amino acids, corresponding to a polypeptide of molecular weight 39605, in good agreement with estimates from SDS gels. The M protein has an amino acid sequence that is both hydrophobic and highly basic. The NDV M protein has sequence homologies to the M proteins of Sendai, measles, canine distemper and respiratory syncytial viruses.

Amino Acid Sequence↗
PubMed1991

Characterization of Newcastle disease viruses isolated from pigeons in Italy.

The authors report the results of characterization studies of three strains of Newcastle disease virus (NDV) (two isolated from pigeons and one from chickens). The plaque cloning of the viruses, showed that each NDV strain consists of different clones of genetically mixed viral populations. The pigeon NDV isolates were classified as lentogenic using mean death time (MDT) determination; while the intracerebral pathogenicity index (ICPI) was the same as the velogenic NDV strain.

Animals↗
PubMedDate not supplied

Newcastle disease virus surveillance in Argentina: use of reverse transcription-polymerase chain reaction and sequencing for molecular typification.

Newcastle disease virus (NDV) remains a major pathogen of poultry where highly virulent strains require reporting to the Office of International Epizootes. NDV is a paramyxovirus existing as different strains classified on the basis of severity of the disease they cause. The present study was conducted in Argentina to determine the prevalence of highly virulent velogenic NDV strains in commercial poultry farms. Tracheal and cloacal swabs from 693 flocks, representing 14% of the broiler production, were collected and pooled. A pool amplified twice in embryonated eggs presented a limited hemagglutination titer. We performed reverse transcription coupled to polymerase chain reaction to amplify fusion and matrix protein gene sequences of the isolate and the strain Trenque Lauquen, isolated in Argentina during an outbreak in 1970-71 and previously characterized as velogenic viscerotropic by biological methods. The amino acid sequences were deduced from nucleotide sequences of the amplification products and the pathotype predicted according to the sequences obtained. From the samples analyzed, we found only one type of NDV, being the isolate identified as lentogenic NDV. This strain is probably the one used in vaccination of flocks where that sample was obtained. These data have allowed us to consider a velogenic NDV-free status in Argentina's commercial poultry.

Amino Acid Sequence↗
PubMed1991

Effect of carrageenan on humoral and protective immune responses of chicks vaccinated with Newcastle disease virus.

The effect of carrageenan (CGN) treatment on the generation of humoral and protective immune response to Newcastle disease virus vaccine was investigated in chicks. Carrageenan treatment significantly impaired the primary humoral immune response in vaccinated chicks. Further, the protective immune response in CGN-treated chicks was only 50% of that in the control vaccinated chicks. However, the post-challenge response was not altered by drug treatment. Indomethacin, a potent inhibitor of prostaglandin synthesis, was able to reverse the immunosuppressive effect of CGN. This finding suggests that prostaglandins might be involved in CGN-mediated immunosuppression in chicks.

Animals↗
PubMedDate not supplied

Effect of Newcastle disease virus on ocular and paraocular tissues in experimentally inoculated chickens.

The effects of a mesogenic strain of Newcastle disease virus on ocular and paraocular structures were studied in 10-to-12-week-old chickens inoculated conjunctivally, intraocularly, intracerebrally, or intravenously. Paraffin-embedded tissues were examined by light and fluorescent microscopy using conventional staining and immunohistochemistry. Lesions were most severe in intraocularly inoculated chickens, where a marked iridocyclochoroiditis was evident from 8-12 hours up to 21 days postinfection. Viral antigens were detected in the iris, ciliary body, Schlemm's canal, and occasionally the lens, choroid, and base of the pecten. Although optic neuritis and iridocyclochoroiditis were found in intracerebrally inoculated birds, no viral antigens were detected in the optic nerve.

Animals↗
PubMed1981

Temperature-sensitive mutant of Newcastle disease virus which has an altered nucleocapsid-associated protein.

Analysis of six temperature-sensitive (ts) mutants of Newcastle disease virus (NDV) representing each of six complementation groups by both SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional gel electrophoresis revealed that in only one mutant was there an alteration in the isoelectric point of a protein. This altered protein was the nucleocapsid-associated protein, NAP. In addition, the mobility of the haemagglutinin-neuraminidase protein, HN, was decreased on non-reduced SDS-PAGE in this mutant. All independent ts+ clones derived from this mutant had normal NAP but HN protein migrated at the decreased rate. Haemagglutinating activity of wide-type (ts+) and ts virions was equally thermostable. Wild-type and ts+ clones derived from this ts mutant were RNA(+) at both permissive and non-permissive temperatures, whereas the ts mutant was RNA(-) at the non-permissive temperature. This ts mutant appears to be a double mutant in both HN and NAP genes, the latter only being a temperature-sensitive lesion which affects virus-directed RNA synthesis.

Capsid↗
PubMed1988

Identification of haemagglutinin-neuraminidase antibody binding sites by Western blot analysis of antibody-resistant mutants and partial digest fragments of Newcastle disease virus.

A collection of monoclonal antibodies (MAbs) which react with the haemagglutinin-neuraminidase (HN) protein of Newcastle disease virus (NDV) has been used to isolate MAb-resistant mutants of the Beaudette C strain of NDV. The patterns of cross-reactivity of the HN proteins of these mutants against the collection of MAbs determined by Western blotting allowed the MAbs to be sorted into different groups. Protease V8 partial digest fragments of purified wild-type virions and subsequent reaction against the collection of MAbs using Western blotting provided an alternative method of grouping MAbs which broadly agreed with the former method. Chemical cleavage of the HN protein at aspartate-proline bonds followed by Western blotting of the fragments allowed the approximate position of certain MAb binding sites to be determined.

Antibodies, Monoclonal↗
PubMedDate not supplied

Observations on the transmissibility of lentogenic strains of Newcastle disease virus: significance of variables.

Virus strain and age of chicken influenced the transmissibility of lentogenic strains of Newcastle disease virus (NDV). The ability of LaSota, B1, V4, CT, F, and Ulster strains to spread from cages of oronasally inoculated chickens to adjacent cages of susceptible chickens was assessed by virus isolation, serology, and immunity to challenge with virulent NDV. Although all inoculated chickens were immune to challenge, the immunity of contact chickens ranged from 100% for LaSota and CT strains to 0% for Ulster strain. The transmissibility of B1 and V4 strains for chickens 1, 4, 8, and 16 weeks old was assessed by within-cage contact infection, exposure to contaminated food and water containers, and exposure to air from infected chickens. Serology and immunity to challenge with virulent virus were used as criteria. Differences in transmissibility were observed for the strain of virus used, route of exposure, and age of chickens. Care must be used in interpreting the significance of strain differences until the effect of variables can be minimized by further improvements in design of the test procedure.

Administration, Intranasal↗
PubMed2002

Chimeric Newcastle disease virus nucleocapsid with parts of viral hemagglutinin-neuraminidase and fusion proteins.

The nucleocapsid (NP) protein of Newcastle disease virus (NDV) self-assembled in Escherichia coli as ring-like and herringbone-like particles. Several chimeric NP proteins were constructed in which the antigenic regions of the hemagglutinin-neuraminidase (HN) and fusion (F) proteins of NDV, myc epitope, and six histidines (a hexa-His tag) were linked to the C-terminus of the NP monomer. These chimeric proteins were expressed efficiently in soluble form in E. coli as detected by Western blot analysis. Electron microscopy of the purified products revealed that they self-assembled into ring-like particles. These chimeric particles exhibited antigenicity of the myc epitope, suggesting that the foreign sequences were exposed on the surface of the particles. Chickens inoculated with the chimeric particles mounted an immune response against NDV, suggesting the possibility of use of the ring-like particle as a carrier of immunogens in subunit vaccines and immunological reagents.

Animals↗
PubMed1975

Thermostability of Newcastle disease virus strains of different virulence.

The thermal inactivation rate constant for infectivity of a total of 24 Newcastle disease virus (NDV) strains was determined at 50 degrees and 56 degrees C. The greater part of the examined NDV strains were found, irrespective of virulence properties, to be thermostable, since the loss of infectivity titre did not exceed 2 logarithmic orders after exposure at 50 degrees C for 60 minutes. Thermostable (I+) and thermolabile (I minus) strains with respect to infectivity were uniformly encountered among the 12 avirulent (lentogenic) and 10 fully virulent (velogenic) strains studied. Strains with both heat stable (Ha+) and heat labile (Ha minus) haemagglutinin were found in the lentogenic group, whereas haemagglutinins of all examined velogenic strains were heat stable. On the basis of I/Ha character, i.e. combination of thermosensitivity of infectivity and haemagglutinin, all examined strains could be classified into one of three categories: I minus Ha minus (8 lentogenic strains), I+Ha+ (4 lentogenic and 7 velogenic strains) and I minus Ha+ (2 mesogenic and 3 velogenic strains). The possible fourth combination, I+Ha+ minus, was not encountered among the strains studied. Determination of the I/Ha character may be helpful in clarifying the origin of a lentogenic NDV strain.

Animals↗