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[In vivo persistence of plaque mutants of viscerotropic Newcastle disease virus].

Studied was the persistance of 3 plaque clones of a velogenic viscerotropic Newcastle disease virus in nonimmune birds. It was found that the first plaque clone (4 mm dia of plaques) possessed higher virulence than that of the parental virus and killed all inoculated birds for 2 to 5 days. Plaque clone II (2.5 mm dia of the plaques) caused up to 30 per cent mortality, and plaque clone III (1.5 mm dia of the plaques) did not bring about death but a transient disease only. Birds inoculated with II and III plaque clones, at identical serologic response, were investigated virologically on the 40th and 60th day post infection. By means of the tracheal organ cultures the Newcastle disease virus was isolated only from birds that were inoculated with plaque clone II. No virus was demonstrated via the same method in birds inoculated with plaque clone III. It is believed that the long-term persistance of the Newcastle disease virus reported on a previous occasion was due to a subpopulation identical with plaque clone II.

Animals↗

Unique hemagglutination activity of an isolate of Newcastle disease virus.

The MET95 strain of a lentogenic Newcastle disease virus (NDV) isolated from a broiler in Japan, showed unique hemagglutination (HA) activity. The MET95 strain failed to show HA when examined by rapid glass plate method although they showed HA titer of 1:1,024 by micro-plate method. This unique HA was also observed after the MET95 strain was passaged ten times in chickens. The failure of HA by rapid glass plate method was not shown in any other NDVs examined.

Animals↗

p53-independent endoplasmic reticulum stress-mediated cytotoxicity of a Newcastle disease virus strain in tumor cell lines.

While Newcastle disease virus (NDV) causes serious infections in birds, it is apparently nonpathogenic in mammalian species, including humans. Previous observations and small-scale clinical trials indicated that NDV exerts oncolytic effects. Isolates of NDV were found to have selective affinity to transformed cells. We previously showed that the attenuated NDV strain MTH-68/H causes apoptotic cell death in cultures of PC12 rat pheochromocytoma cells. The aim of the present study was to extend MTH-68/H cytotoxicity testing with human tumor cell lines and to analyze certain biochemical aspects of its oncolytic effect. MTH-68/H was found to be able to kill a wide range of transformed cells by apoptosis. While caspase-8 and caspase-9 are not involved in MTH-68/H-induced apoptosis, activation of caspase-3 and caspase-12 was detected in virus-infected PC12 cells. A human glioblastoma cell line with repressible expression of the p53 protein did not show any difference in MTH-68/H sensitivity in its p53-expressing and p53-depleted states, indicating that the apoptotic process induced by MTH-68/H does not depend on p53. Apoptosis was accompanied by virus replication in two tumor cell lines tested (PC12 cells and HeLa human cervical cells), and signs of endoplasmic reticulum stress (phosphorylation of protein kinase R-like endoplasmic reticulum kinase and eIF2alpha) were also detected in transformed cells. In contrast, proliferation of nontransformed mouse and rat fibroblast cell lines and human primary fibroblasts was not affected by MTH-68/H treatment. MTH-68/H thus selectively kills tumor cell cultures by inducing endoplasmic reticulum stress leading to p53-independent apoptotic cell death.

Animals↗

Fusogenic activity of reconstituted newcastle disease virus envelopes: a role for the hemagglutinin-neuraminidase protein in the fusion process.

Enveloped viruses, such as newcastle disease virus (NDV), make their entry into the host cell by membrane fusion. In the case of NDV, the fusion step requires both transmembrane hemagglutinin-neuraminidase (HN) and fusion (F) viral envelope glycoproteins. The HN protein should show fusion promotion activity. To date, the nature of HN-F interactions is a controversial issue. In this work, we aim to clarify the role of the HN glycoprotein in the membrane fusion step. Four types of reconstituted detergent-free NDV envelopes were used, on differing in their envelope protein contents. Fusion of the different virosomes and erythrocyte ghosts was monitored using the octadecyl rhodamine B chloride assay. Only the reconstituted envelopes having the F protein, even in the absence of HN protein, displayed residual fusion activity. Treatment of such virosomes with denaturing agents affecting the F protein abolished fusion, indicating that the fusion detected was viral protein-dependent. Interestingly, the rate of fusion in the reconstituted systems was similar to that of intact viruses in the presence of the inhibitor of HN sialidase activity 2,3-dehydro-2-deoxy-N-acetylneuraminic acid. The results show that the residual fusion activity detected in the reconstituted systems was exclusively due to F protein activity, with no contribution from the fusion promotion activity of HN protein.

Animals↗

Conformationally sensitive antigenic determinants on the HN glycoprotein of Newcastle disease virus form with different kinetics.

The mature Newcastle disease virus (NDV) hemagglutinin-neuraminidase (HN) protein is a type 2, oligomeric glycoprotein which has seven antigenic sites, sites 1, 2, 3, 4, 23, 12, and 14 (lorio et al., 1989, Virus Res. 13, 245). The folding of the HN protein was explored by characterizing the formation of representative epitopes specific for each of the six of antigenic sites in infected cells. None of these epitopes was present on the nascent, 5-min pulse-labeled HN protein, while all epitopes appeared after a 1- to 2-hr chase. All epitopes formed in the presence of monensin or during a chase at 15 degrees, suggesting that all these determinants appear while the molecule is in the rough endoplasmic reticulum. However, none of the epitopes appeared during chases in the presence of carbonyl cyanide m-chlorophenylhydrazone, suggesting that the formation of all determinants requires ATP. The kinetics of formation of each of the determinants was quantitated in both NDV-infected Cos cells and chick embryo cells. In both cell types, antigenic determinants formed with different kinetics. The epitope specific for site 4 appeared first, followed by the simultaneous appearance of epitopes in sites 1 and 3. The epitope in site 2 appeared next and that in site 23 last. The kinetics of appearance of the epitope in site 14 relative to those in other sites varied with cell type. In chick cells, this epitope appeared with the site 2 epitope, while in Cos cells this epitope appeared with or just after sites 1 and 3 epitopes. Nonradioactive chases at 15 degrees slowed the formation of the antigenic determinants. The disulfide-linked dimer form of the HN protein appeared concomitant with epitopes in sites 1 and 3.

Animals↗

Structural and functional characterization of Newcastle disease virus polycistronic RNA species.

Upon infection, the Newcastle disease virus (NDV) genome is transcribed to produce 18S, 22S, and 35S RNAs (M. Bratt , and W. Robinson, J. Mol. Biol. 23:1-21, 1967). The 22S RNA has been shown to contain 18S sequences and is thought to represent polycistronic transcripts generated by transcriptional readthrough of adjacent genes ( Varich et al., Acta Virol. 23:341-343, 1979). With improved extraction procedures, the 22S RNA was found to represent up to 25% of the total transcription in NDV-infected cells. This RNA was resolved into at least five discrete species on formaldehyde-agarose gels. All but one of these molecules contain 3' polyadenylate sequences but not internal polyadenylate sequences. These transcripts are found on polyribosomes of infected cells, suggesting that they are functional mRNAs.

Animals↗

Isolation and preliminary characterization of temperature-sensitive mutants of Newcastle disease virus.

Temperature-sensitive (ts) mutants of Newcastle disease virus have been isolated and characterized genetically (complementation), biochemically (RNA synthesis) and biologically (fusion from within and hemadsorption). Fifteen of these mutants have been divided into five complementation groups. Groups A (five mutants) and E (one mutant) are ts for RNA synthesis (RNA-) as well as for the other functions. Group B contains four RNA+ mutants of which one is ts for fusion, one for hemadsorption and two for neither function. Group C contains one RNA+ mutant which is a poor cell fuser. Group D contains two RNA+ mutants which are ts for fusion. In addition, two noncomplementing mutants (group BC) fail to complement both group B and group C mutants while exhibiting complementation with mutants in groups A, D, and E.

Cell Fusion↗

Phylogeography and molecular evolution of Newcastle disease virus across a century of global surveillance.

Newcastle disease virus (NDV) remains one of the most economically important avian pathogens worldwide, causing recurrent outbreaks in poultry despite decades of vaccination and disease control efforts. Since the first reported outbreak of NDV a hundred years ago, numerous molecular epidemiological studies have been conducted globally across diverse geographic and production settings. Following a century of NDV circulation and evolution, the present study aimed to compile all publicly available NDV sequence data and perform a comprehensive global analysis of the genetic diversity, phylogenetic relationship, and global spatiotemporal distribution of NDV over a 100-year timescale. All publicly available NDV complete genome and full-length fusion (F) gene sequences were retrieved from GenBank up to February 2026. Following rigorous quality control, phylogenetic analyses were performed based on complete genomes and F gene datasets. Phylogenetic analysis identified two genotypes within Class I and 20 genotypes within Class II NDVs, with extensive diversification at the sub-genotype level. Genotype XIII exhibited the greatest sub-genotypic diversity, while genotype VII represented the most globally disseminated genotype, reported across 36 countries. Chronological assessment based on the earliest available reports indicated an increasing number of recognized genotypes from the 1930s to recently described sub-genotypes such as XIII.2.3 and XXII.2.2. Regional diversity analysis revealed the highest genotype diversity in Western Africa, Eastern Asia, and Southern Asia. Comparative residue analysis demonstrated substantial genotype-specific variation within critical functional domains of the fusion protein, including cleavage sites, neutralizing epitopes, and heptad repeat regions. Overall, this study provides the first comprehensive 100-year global overview of NDV evolution and phylogeography. The findings highlight continuous viral diversification, broad geographic dissemination of multiple genotypes, and ongoing molecular variation, emphasizing the need for sustained genomic surveillance and periodic evaluation of vaccine compatibility with emerging NDV genotypes.

100-years of data↗

[Nonspecific non-reactivity in mice induced by joint administration of Newcastle disease virus and cyclophosphamide].

The immunomodulating effect of Newcastle disease virus (NDV) was investigated in vitro and in vivo in mice. NDV was shown to induce a mitogenic effect in splenocytes in vitro. Combined injections of NDV and CP resulted in nonspecific suppression of immunoreactivity in mice. The antibody production and development of delayed type hypersensitivity to sheep erythrocytes were markedly reduced. Injections of NDV alone slightly increased the reactions. The NDV + CP injections led also to a reduction of immune response to thymus-independent antigen, LPS. Thus, the combined injections of NDV and CP led to nonspecific suppression of T and B cell immunity in mice. The mechanisms of this form of anergy require further study.

Animals↗

Characterization of a lentogenic Newcastle disease virus isolated from broiler chickens in Japan.

Newcastle disease virus (NDV), named MET95, was isolated from a non-vaccinated broiler flock in Japan in 1995. The MET95 strain was determined to be a lentogenic NDV. The strain has the properties of eluting rapidly at 4 C and has low thermostability in hemagglutinating activity with chicken erythrocytes. In these studies, no difference could be found between the MET95 strain and the Hitcher B1 vaccine strain. However, the chickens inoculated with the MET95 strain, as well as chickens that they were in contact with, had a much higher hemagglutination-inhibition antibody response than those inoculated with the B1 strain. Accordingly, the MET95 strain is thought to be a promising candidate as a live ND vaccine strain. In Japan, this is the first report on the isolation of lentogenic NDV from chickens since the paper on the Ishii strain isolated in 1966.

Animals↗

Evidence for mixed membrane topology of the newcastle disease virus fusion protein.

The synthesis of the Newcastle disease virus (NDV) fusion (F) protein in a cell-free protein-synthesizing system containing membranes was characterized. The membrane-associated products were in at least two different topological forms with respect to the membranes. The properties of one form were consistent with the expected membrane insertion as a classical type 1 glycoprotein. This form of the protein was fully glycosylated, and sequences amino terminal to the transmembrane domain were protected from protease digestion by the membranes. The second form of membrane-associated F protein was partially glycosylated and partially protected from protease digestion by the membranes. Protease digestion resulted in a 23-kDa protease-protected polypeptide derived from F2 sequences and sequences from the amino-terminal end of the F1 domain. Furthermore, a 10-kDa polypeptide derived from the cytoplasmic domain (CT) was also protected from protease digestion by the membranes. Protease resistance of the 23- and 10-kDa polypeptides suggested that this second form of F protein inserted in membranes in a polytopic conformation with both the amino-terminal end and the carboxyl-terminal end translocated across membranes. To determine if this second form of the fusion protein could be found in cells expressing the F protein, two different approaches were taken. A polypeptide with the size of the partially translocated F protein was detected by Western analysis of proteins in total-cell extracts of NDV strain B1 (avirulent)-infected Cos-7 cells. Using antibodies raised against a peptide with sequences from the cytoplasmic domain, CT sequences were detected on surfaces of F protein-expressing Cos-7 cells by immunofluorescence and by flow cytometry. This antibody also inhibited the fusion of red blood cells to cells expressing F and HN proteins. These results suggest that NDV F protein made both in a cell-free system and in Cos-7 cells may exist in two topological forms with respect to membranes and that the second form of the protein may be involved in cell-cell fusion.

Amino Acid Sequence↗

Rapid serological profiling by enzyme-linked immunosorbent assay. III. Simultaneous measurements of antibody titers to infectious bronchitis, infectious bursal disease, and Newcastle disease viruses in a single serum dilution.

The present paper describes a method for measuring enzyme-linked immunosorbent assay (ELISA) antibody titers to infectious bronchitis (IB), infectious bursal disease (IBD), and Newcastle disease (ND) viruses from a single serum dilution using the same assay procedures and analyses. A regression line equation generated for predicting NDV ELISA antibody titers at a single serum dilution was successfully used to predict ELISA antibody titers to IB and IBD antigen preparations at the same test sera dilutions. Duplicate samples of 46 field and control sera were transferred simultaneously, with the aid of a replica plating device, from a reservoir plate to three different test plates, which were coated separately with IB, IBD, and ND antigens. After similar indirect ELISA parameters were conducted for each antigen, raw absorbance values from duplicate tests and controls were transmitted directly from an ELISA reader to a microcomputer, which subsequently processed average corrected absorbance values directly into individual ELISA antibody titer for each test antigen. The computer output listed individual field and control sample titers to each antigen, as well as graphing the relative ELISA titer distribution of the entire test group to each antigen. In a comparative study, test sera were evaluated for ELISA antibody titer to NDV, IBV, and IBDV by this method in three separate assays. Consistent titer values were obtained in all assays, as 96% of the 414 replicate titers evaluated varied less than twofold.

Animals↗

Haemagglutinating activity of the lentogenic Newcastle disease virus strain MET95.

Using the rapid glass plate method, the Newcastle disease virus strain MET95 showed much weaker haemagglutination (HA) activity for chicken erythrocytes than 69 other Newcastle disease viruses, including 56 field strains isolated from chickens reared in Japan between 1988 and 2001. Using erythrocytes from other avian species, only the MET95 strain failed to show HA activity for erythrocytes from ducks, geese or pigeons. The haemagglutinin-neuraminidase protein of the MET95 strain was shown to have unique substitutions of isoleucine for thereonine and leucine at amino acide residues 216 and 552. It is suggested that these two substitutions might relate to the unique HA activity of the MET95 strain. This HA activity may be a useful marker for this strain.

Amino Acid Sequence↗

Recombinant fowlpox viruses coexpressing chicken type I IFN and Newcastle disease virus HN and F genes: influence of IFN on protective efficacy and humoral responses of chickens following in ovo or post-hatch administration of recombinant viruses.

We have constructed recombinant (r) fowl pox viruses (FPVs) coexpressing chicken type I interferon (IFN) and/or hemagglutinin-neuraminidase (HN) and fusion (F) proteins of Newcastle disease virus (NDV). We administered rFPVs and FPV into embryonated chicken eggs at 17 days of embryonation or in chickens after hatch. Administration of FPV or rFPVs did not influence hatchability and survival of hatched chicks. In ovo or after hatch vaccination of chickens with the recombinant viruses resulted in protection against challenge with virulent FPV and NDV. Chickens vaccinated with FPV or FPV-NDV recombinant had significantly lower body weight 2 weeks following vaccination. This loss in body weight was not detected in chickens receiving FPV-IFN and FPV-NDV-IFN recombinants. Chickens vaccinated with FPV coexpressing IFN and NDV genes produced less antibodies against NDV in comparison with chickens vaccinated with FPV expressing NDV genes.

Animals↗

Lectin binding and the carbohydrate moieties present on Newcastle disease virus strains.

Lectin-binding profiles were developed for 14 strains of Newcastle disease virus in order to determine the carbohydrate moieties associated with hemagglutination and to establish whether there are any associations between the carbohydrates present on the virus envelope and virulence. All strains of Newcastle disease virus were bound by concanavalin A. Other lectins bound the viruses differentially, but there was no pattern of binding that could be associated with viral virulence. The binding of virus by Lens culinaris lectin was associated with the elution rate of the virus from chicken erythrocytes. Strains that elute rapidly from chicken erythrocytes were not bound by Lens culinaris lectin. The sugar alpha-D-N-acetylglucosamine inhibited the adsorption of Lens culinaris lectin to the strains that were "slow eluters" from chicken erythrocytes.

Acetylgalactosamine↗