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PubMed1995

Australian studies on Newcastle disease virus. The French heritage.

Eric French contributed greatly to the early Australian studies on Newcastle disease virus, producing the foundations on which subsequent Australian studies were based. In 1964 he conducted the first major serological survey for Newcastle disease in the Australian poultry flock, and showed that the pathotypes of the virus recognised at that time were not present. After the isolation of strain V4 in 1966, he initiated some of the first studies on the nature of this stain. In particular, he demonstrated the avirulence of this virus, its ability to infect chickens when delivered orally with food and its potential utility as a vaccine. Subsequent studies by other workers included the development of strain V4 as a conventional vaccine and as a vaccine suitable for use in village chickens.

Animals↗
PubMedDate not supplied

Nucleotide sequence and phylogenetic analysis of Newcastle disease virus isolates from recent outbreaks in Taiwan.

Portions of the hemagglutinin neuraminidase (HN) gene of Newcastle disease virus (NDV) isolates from two recent outbreaks were sequenced to investigate epidemiology of this disease in Taiwan. These NDV isolates were all viscerotropic velogenic according to the clinical lesions produced in chickens. Sequence data were obtained from 14 NDV isolates (12 from 1995 and 2 from 1984). All isolates differed in their nucleotide sequences (from 0.3 to 15.3%), and represented potentially different strains of NDV. Phylogenetic analysis revealed that these isolates are closely related to viruses isolated from Japan and Malaysia. Some viruses isolated in 1995 appeared to evolve from viruses isolated in 1984. The results suggest that the 1995 outbreak of Newcastle disease (ND) in Taiwan may have been caused by multiple strains of velogenic NDV that have cocirculated in Taiwan for some time. Moreover, NDV isolates from racing pigeons were very similar to isolates from chickens in the same period, suggesting that both domestic and free-living birds were involved in the spread of ND in Taiwan.

Amino Acid Sequence↗
PubMedDate not supplied

The effects of the multiplicity of infection on viral subpopulations during passage of Newcastle disease viruses.

When mixtures of two clones of the Italy-Milano strain of Newcastle disease virus were inoculated so that one clone was present in quantities 20 times greater than the other, maintenance of that population ratio through six serial passages in 10-day-old embryonating chicken eggs depended on the multiplicity of infection of the first passage. The initial ratio was preserved when the eggs were inoculated at low multiplicity, 10(1) virions inoculated, but not at high multiplicity, 10(6) virions inoculated. In the latter situation, the clear plaque clone, which comprised 1/20 of the initial population of the inoculum, increased in numbers to parity with the red plaque clone after only six serial passages in embryonating eggs. Explanations for the competitive advantage of the clear plaque clone at high multiplicity of infection are discussed.

Animals↗
PubMedDate not supplied

Antibody response to strain combinations of Newcastle disease virus as measured by hemagglutination-inhibition.

Differences in antibody response to three Newcastle disease virus (NDV) strains--B-1, LaSota, and Ulster--were investigated using the hemagglutination-inhibition (HI) micro-titer test in chickens hatched from ND-immune and unimmune flocks. When used singly as primary vaccines, the Ulster strain stimulated the lowest antibody response of the three in both immune and unimmune (susceptible) chickens. Subgroups of each of the primary-vaccinated groups were revaccinated with each of the three strains. Ulster-vaccinated chicks, revaccinated with Ulster, gave the poorest booster response. All other revaccination combinations gave a significant titer increase, though some were better than others. It is suggested that the Ulster strain as primary vaccine followed by booster does of B-1 or LaSota will induce a higher antibody response (i.e., immunity) in susceptible chicken populations with less risk of a post-vaccination reaction.

Animals↗
PubMed2004

Conformational changes of Newcastle disease virus envelope glycoproteins triggered by gangliosides.

We have investigated the conformational changes of Newcastle disease virus (NDV) glycoproteins in response to receptor binding, using 1,1-bis(4-anilino)naphthalene-5,5-disulfonic acid (bis-ANS) as a hydrophobicity-sensitive probe. Temperature- and pH-dependent conformational changes were detected in the presence of free bovine gangliosides. The fluorescence of bis-ANS was maximal at pH 5. The binding of bis-ANS to NDV was not affected by chemicals that denature the fusion glycoprotein, such as reducing agents, nor by the presence of neuraminidase inhibitors such as N-acetyl neuramicic acid. Gangliosides partially inhibited fusion and hemadsorption, but not neuraminidase hemagglutinin-neuraminidase glycoprotein (HN) activity. A conformational intermediate of HN, triggered by the presence of gangliosides acting as receptor mimics, was detected. Our results indicate that, upon binding to free gangliosides, HN undergoes a certain conformational change that does not affect the fusion glycoprotein.

Anilino Naphthalenesulfonates↗
PubMed1986

Inhibition of lymphocyte mitogenesis in mice infected with Newcastle disease virus: viral interference with the interleukin system.

Spleen cells from mice infected with Newcastle disease virus (NDV) fail to proliferate when cultured with allogeneic cells or with concanavalin A (Con A). This failure is not due to impairment of interleukin-1 (IL-1) production or to a lack of accessory cell function as stimulator cells from NDV-infected mice induce DNA synthesis in the mixed lymphocyte reaction. However, spleen cells from NDV-infected mice fail to produce detectable amounts of interleukin-2 (IL-2) when stimulated with mitogenic doses of Con A and do not respond to exogenous IL-2-containing preparations. Furthermore, absorption experiments suggest that cells from NDV-infected mice fail to bind appreciable amounts of exogenous IL-2. All these events seem to be infection-dependent, as cells from mice injected with ultraviolet-inactivated NDV (UV-NDV) behave normally.

Animals↗
PubMed1978

Transcription and translation of Newcastle disease virus mRNA's in vitro.

Transcription directed in vitro by Triton-activated Newcastle disease virus (NDV) was stimulated and prolonged by the presence of cytoplasmic extracts of animal cells. The RNA products closely resembled those of NDV transcription in vivo by several criteria: binding to oligodeoxythymidylic acid-cellulose, the mobility and relative abundance of each major band resolved by polyacrylamide gel electrophoresis, and the ability to direct the accurate cell-free synthesis of polypeptides corresponding to the NDV proteins HN, F0/F1, NP, and M. Synthesis of a novel polypeptide related to NP but of higher apparent molecular weight was also detected. These results indicated that cell-free transcription under these conditions was a close facsimile of NDV transcription in vivo. In addition, both in vitro and in vivo, NDV polypeptides were synthesized in nonequimolar amounts which reflected the order of the genes in the transcriptional map: NP, F0, M, (47K, HN), L. Strains AV and HP, virulent strains which have differences in biological activities, exhibited differences in the polypeptides synthesized in infected cells and in cell-free systems.

Cell-Free System↗
PubMed1978

[Cytopathic effect of the Newcastle disease virus on the BHK-21 cell line].

The cytopathic effect (CPE) of the Newcastle disease virus (NDV) in the BNK-21 permenant cell line is studied in a series of experiments. The CPE is assessed as a percentage of hemadsorption and polykaryocytosis, and the dynamics of the viral replication is followed up with the aid of the immunofluorescent method. The effect of the cyclohexamide protein inhibitor of the replication of various NDV strains, on the CPE, respectively, is likewise studied. Five vellogenic, one mesogenic and one lentogenic strains are used in the experiments. It is found that through the immunofluorescent method the viral replication is delayed some three hrs in the case of the La Sota lentogenic strain in comparison with the other strains used. The direct relationship, already established, between the virulence of the NDV and the percentage of haemadsorption and polykaryocytosis is confirmed also in the BNC-21 heterologous cellular tissue. On the basis of this relationship, a rapid typing method of virulent from avirulent field NDV strains is suggested. The blocking effect of the cyclohexamide protein inhibitor on the CPE in NDV is demonstrated in the BNK-21 line, as a result of the blocking of the viral specific protein synthesis.

Animals↗
PubMedDate not supplied

Biological and molecular characterization of Newcastle disease virus (NDV) field isolates with comparisons to reference NDV strains.

Fifty-seven Newcastle disease virus (NDV) isolates from chickens, turkeys, a rhea, a parrot, and an anhinga were pathotyped and characterized by monoclonal antibody (mAb) inhibition profile, elution rate, and hemagglutinin thermostability. Nucleotide sequence analysis of portions of the fusion protein and matrix protein genes of the parrot isolate was done for comparison with prior sequence analysis of the anhinga isolate and NDV reference strains. Seven of the 43 chicken isolates were recovered from flocks in Canada. The remaining isolates, including 11 from turkeys, were isolated in the United States. All isolates except that of the anhinga were of low virulence by mean death time in embryos, intracerebral pathogenicity index, and/or intravenous pathogenicity index procedures and were classified as lentogens. The anhinga isolate was more virulent than the other strains and was pathotyped as a mesogen. However, nucleotide sequence analysis of the anhinga isolate had revealed a homology with the virulent cormorant isolates of 1992 rather than the classical U.S. mesogens characterized by the Roakin strain. Variability was evident among the lentogenic isolates. Two isolates from turkeys had mAb profiles that differed from B1 and La Sota reference and vaccine strains, and 38% (21/56) of the isolates had more thermostable hemagglutinins than those reference strains. There was no evidence that any of the isolates from poultry were more virulent than the lentogenic pathotype.

Animals↗
PubMed1993

In vitro expansion and analysis of T lymphocyte microcultures obtained from the vaccination sites of cancer patients undergoing active specific immunization with autologous Newcastle-disease-virus-modified tumour cells.

In order to understand further the effects of Newcastle-disease-virus(NDV)-modified tumour vaccines we investigated the feasibility of isolating lymphocytes from the site of injection of patients undergoing postoperative active specific immunization (ASI) with autologous NDV-modified tumour cells. Delayed-type-hypersensitivity(DTH)-like reactions from five cancer patients were surgically removed, minced and the tissue particles were digested with collagenase and DNase. Lymphoid cells recovered were expanded in a highly efficient limiting-dilution analysis system optimized for T cell growth [Moretta et al. (1983) J Exp Med 157: 743] and lymphocyte microcultures (clonal probability > 0.8) could be grown for up to 1 year. Analysis of the microcultures for phenotype and function showed that the majority were positive for CD4 (92%) and TCR alpha beta (96%). Concanavalin-A-induced production of interleukin-2 (IL-2), IL-6, interferon gamma and tumour necrosis factor alpha was detected in more than 70% of the microcultures. Lectin-dependent cytotoxicity was only very rarely observed. The general characteristics of the microcultures obtained support the notion of a DTH-like reaction taking place at the site of tumour cell challenge. The possibility of in vitro expansion and cultivation of T lymphocytes from ASI vaccination sites should help to elucidate further the role of these cells in active specific immunization against autologous tumour cells.

Cells, Cultured↗
PubMed1980

Maturation of the hemagglutinin-neuraminidase and fusion glycoproteins of two biologically distinct strains of Newcastle disease virus.

We have compared the glycoproteins of two biologically distinct virulent strains of Newcastle disease virus. Cells infected by either strain AV or HP produce infections virions and the cellular surfaces have hemadsorbing activity; however, only cells infected by strain AV undergo fusion from within. We fractionated chicken embryo cells and monitored the incorporation of radioactive proteins into virions to study the sites of synthesis, cellular locations, and kinetics of virion assembly for the hemagglutinin-neuraminidase (HN) and the fusion glycoproteins of each strain. We found that the HN glycoprotein of both strains was synthesized on rough endoplasmic reticulum (ER), accumulated in low-density membranes derived from smooth ER and the plasma membrane, and appeared in virions after a 30-min delay. The fusion glycoprotein of both strains was synthesized as a precursor in rough ER and subsequently processed to the active form. However, the sites of accumulation of the fusion glycoproteins were strain-dependent. The larger subunit F1 of the fusion glycoprotein of strain AV was detected in subcellular fractions enriched for plasma membranes, while that of strain HP accumulated in denser fractions which contained internal membranes. This result suggests that differential compartmentation of viral glycoproteins may influence the expression of biologic activities such as fusion on cellular surfaces. Despite their different sites of accumulation, the F1 glycoproteins or F-related polypeptides of similar size were assembled into virions of both strains. The kinetics of incorporation of this protein into virions appear to be too rapid for migration through internal membrane systems. Furthermore, tunicamycin did not block the incorporation of F-related polypeptides into virions. A hypothesis is presented to explain the unusual behavior of the F-related proteins.

Animals↗
PubMed2002

Pathogenesis of six pigeon-origin isolates of Newcastle disease virus for domestic chickens.

The pathogenesis of six pigeon-origin isolates of Newcastle disease virus (NDV) was investigated in chickens. Four isolates were previously defined as the variant pigeon paramyxovirus 1 (PPMV-1), and two isolates were classified as avian paramyxovirus 1 (APMV-1). Birds inoculated with PPMV-1 isolates were euthanatized, and tissue samples were collected at 2, 5, and 10 days postinoculation (DPI). Birds inoculated with APMV-1 isolates died or were euthanatized, and tissue samples were collected at 2, 4, and 5 DPI. Tissues were examined by histopathology, immunohistochemistry (IHC) for the presence of NDV nucleoprotein, and in situ hybridization (ISH) for the presence of viral mRNA for the matrix gene. Spleen sections were stained by the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay and by IHC using an anti-active caspase-3 antibody (IHC-Casp) to detect apoptotic cells. Brain sections of PPMV-1-infected birds were examined by IHC to detect T and B lymphocytes and glial fibrillary acidic protein (GFAP). Histologically, birds inoculated with PPMV-1 isolates had marked lesions in the heart and brain. Presence of viral nucleoprotein and viral mRNA in the affected tissues was confirmed by IHC and ISH, respectively. Numerous reactive astrocytes were observed in brain sections stained for GFAP Among all the isolates, the IHC-Casp demonstrated that apoptosis was very prominent in the ellipsoid-associated cells of the spleen at 2 DPI. Results of the TUNEL assay indicated that apoptotic cells were prominent at 5 DPI and were more randomly distributed. The clinical signs and gross and histopathologic changes observed in the APMV-1-infected birds were characteristic of an extensive infection with highly virulent NDV evident by IHC.

Animals↗
PubMed1994

The role of the individual cysteine residues in the formation of the mature, antigenic HN protein of Newcastle disease virus.

The amino acid sequence of the hemagglutinin-neuraminidase (HN) glycoprotein of Newcastle disease virus (NDV) has 14 cysteine residues, two of which are variably present in the sequences of the HN proteins of different strains of NDV while the rest are absolutely conserved. The role of each residue in the formation of the mature, oligomeric structure of the HN protein was assessed by characterizing proteins with mutations in each of the cysteine residues by Western analysis, immunoprecipitation with conformationally sensitive antibodies, immunofluorescence, and sedimentation on sucrose gradients. Proteins with mutations in the first cysteine (amino acid 6) or the second cysteine (amino acid 123), the nonconserved cysteine residues, formed antigenically mature oligomers which were transported to the cell surface like wild type. Protein with a mutation at cysteine 2 did not, however, form covalently linked oligomers demonstrating that it is this residue that is responsible for intermolecular disulfide bonds in the mature oligomer. Proteins with mutations in cysteine 3 (amino acid 172) or cysteine 5 (amino acid 196) formed proteins with all antigenic sites except one, site 23. These mutant proteins formed disulfide-linked dimers and were efficiently transported to the cell surface. They did not, however, sediment on gradients like the wild-type protein. They were also defective in the biological activities associated with the wild-type protein. Proteins with mutations in cysteines 4 (amino acid 186), 6 (amino acid 238), 7 (amino acid 247), 8 (amino acid 251), 13 (amino acid 531), or 14 (amino acid 542) contained no mature antigenic sites but formed noncovalently linked oligomers. Proteins with mutations in cysteines 9 (amino acid 344), 10 (amino acid 455), 11 (amino acid 461), or 12 (amino acid 465) formed proteins with antigenic site 4 but no other mature antigenic sites. These mutant proteins also formed noncovalently linked oligomers. These results suggest that mutations in different cysteine residues block the maturation of the HN protein at different stages.

Animals↗
PubMed2006

Newcastle disease virus (strain Herts 33/56) in tissues and organs of chickens infected experimentally.

Six-week-old susceptible specific pathogen free chickens were infected intranasally with the virulent Newcastle disease virus strain Herts 33/56 and the levels of virus present in blood, faeces, breast muscle, leg muscle and a pool of heart/kidney/spleen were estimated in birds killed humanely at each day post inoculation. Highest titres were recorded at day 4 post inoculation when titres of virus were 10(6) median egg infectious doses (EID50)/g in the heart/kidney/spleen pool, 10(4.2) EID50/g in the leg muscle and 10(4) EID50/g in the breast muscle and faeces. A median oral infectious dose of Newcastle disease virus strain Herts 33/56 for 3-week-old chickens was estimated to be equivalent to 10(4) EID50.

Animals↗
PubMed1979

Interferon induction with Newcastle disease virus in FS-4 cells: effect of priming with interferon and of virus inactivating treatments.

Inoculation of human FS-4 cells with Newcastle disease virus (NDV) resulted in the induction of two distinct interferon responses, one that peaked at about 5 hr (early response) and one that reached a maximum between 10 to 24 hr after inoculation (second response). The early interferon response was enhanced by previous treatment of the cells with interferon (priming), whereas the second response decreased after interferon treatment in a dose-dependent manner. The early response diminished with decreasing multiplicities of infection, the magnitude of the second response in unprimed cells was relatively independent of the dose of NDV employed. The early interferon response was sensitive to inhibition by actinomycin D for only 1 hr after inoculation. In marked contrast, the second response remained sensitive to inhibition by actinomycin D until 12 hr after inoculation. The ability of NDV to induce the second response was greatly diminished by irradiation of the virus with ultraviolet light or by its treatment with hydroxylamine, whereas the ability to stimulate the early response was relatively resistant to these virus-inactivating treatments. Treatment of NDV with hydroxylamine abolished the virus to induce the second response at the same rate as it destroyed infectivity. The results suggest the existence of at least two distinct mechanisms of interferon induction by NDV; the early response is triggered either by a virion component or by a product of primary transcription, whereas induction of the second response requires the expression of some functions of the virus not needed for triggering the early response.

Animals↗