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PubMed1990

A temperature-sensitive mutant of Newcastle disease virus defective in intracellular processing of fusion protein.

A temperature-sensitive mutant (ts3) of Newcastle disease virus was physiologically characterized. All major viral structural proteins were synthesized at the permissive (37 degrees C) and nonpermissive (42 degrees C) temperatures, but the fusion (F) glycoprotein was not cleaved at 42 degrees C. In immunocytochemical electron microscopy, the F protein was abundant in the rough endoplasmic reticulum but not in cytoplasmic membrane at 42 degrees C. Noninfectious hemagglutinating virus particles containing all major structural proteins except the F protein were released at 42 degrees C from infected cells. We concluded that the defect in ts3 resides in the intracellular processing of the F protein.

Animals↗
PubMed1992

Effects of the components of Newcastle disease virus on the structural order of lipid assemblies.

The membrane M-protein of Newcastle disease virus is localized directly beneath the lipid bilayer. Although this protein is the major constituent of the virus, its structural relationship to the lipid or to the other viral component hemagglutinin-neuraminidase, the so called HN-glycoprotein, is still unknown. The effects of either M-protein alone or both M-protein and HN-glycoprotein on the lipid assemblies in reconstituted liposomes were determined by differential polarized phase fluorometry, steady-state fluorescence anisotropy and emission lifetime measurements. It is demonstrated that the degree of rotation of fluorophores in reconstituted liposomes is restricted by the molecular packing of lipids in the bilayer and this in turn can be correlated with the structural order of the lipids in the membrane. The experimental results show that the structural order parameters calculated from the fluorescence measurements are strongly influenced by the presence of both M-protein and HN-glycoprotein in the lipid assemblies.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed2006

The pathogenesis of Newcastle disease: a comparison of selected Newcastle disease virus wild-type strains and their infectious clones.

The effect of mutations of Newcastle disease virus (NDV) fusion (F) gene, hemagglutinin-neuraminidase (HN) gene, and phosphoprotein (P) gene and HN chimeras between the virulent Beaudette C and low virulence LaSota strains on pathogenesis and pathogenicity was examined in fully susceptible chickens. A virulent F cleavage site motif within a LaSota backbone increased pathogenicity and severity of clinical disease. A LaSota HN within a Beaudette C backbone decreased pathogenicity indices and disease severity. A Beaudette C HN within a LaSota backbone did not change either pathogenicity indices or severity of disease in chickens. Loss of glycosylation at site 4 of the HN or modified P gene of Beaudette C decreased pathogenicity indices and caused no overt clinicopathologic disease in chickens. Both pathogenicity indices and clinicopathologic examination demonstrated that the F, HN, and P genes of NDV collectively or individually can contribute to viral virulence.

Animals↗
PubMedDate not supplied

Selection of thermostable Newcastle disease virus progeny from reference and vaccine strains.

In a study of low-virulence Newcastle disease virus (NDV) isolates from poultry, 38% of the isolates had a more thermostable hemagglutinin than the lentogenic reference strains B1 and La Sota or live vaccines derived from those strains. Whether those strains with a more thermostable hemagglutinin are truly indigenous or whether they could have originated from vaccines used in the flocks was unknown. Seven monovalent NDV vaccines of B1 or La Sota type and reference B1 and La Sota strains were heat treated at 56 C to select variants more thermostable than the parent virus. Four thermal treatment cycles were completed, and virus propagated from the second and fourth heat treatments was assayed for changes in thermostability and antigenicity. The hemagglutinin thermostability of all vaccine and reference strain variants increased from the initial < or =10 min to > or =120 min after four treatments. Antigenic changes evaluated by hemagglutination inhibition against NDV monoclonal antibodies identified changes in only the heat-treated La Sota strains. The results demonstrate that the field isolates with a more thermostable hemagglutinin could have been derived by selection from the heterogenous NDV populations in vaccine strains and that minor antigenic changes may be a result of that selection.

Animals↗
PubMed2005

Tumor-targeted gene transfer in vivo via recombinant Newcastle disease virus modified by a bispecific fusion protein.

Previously we have demonstrated that a recombinant Newcastle disease virus (NDV) carrying the transgene EGFP can be retargeted to IL-2 receptor positive tumor cells by a bispecific fusion protein alphaHN-IL-2 in vitro. The purpose of the present study was to investigate the specificity and efficiency of gene delivery to tumor cells in vivo via this modified RNA virus. Prior ex vivo infection of murine lymphoma cells by the modified virus resulted in selective EGFP expression in IL-2R+ target tumor cells in vivo. Direct fluorescence microscopy and immunohistology showed viral replication in target positive tumor tissue resulting in much more EGFP expression than in target negative tumor tissue, 24 h after intratumoral injection of the alphaHN-IL-2 modified NDV. A quantitative real-time RT-PCR for EGFP mRNA. confirmed the selective gene expression in IL-2R+ tumor cells. Biodistribution studies showed that EGFP transgene delivery was reduced by 35-100% in liver, spleen, kidney, lung and thymus by the modified virus, while 98% of the transgene was delivered to IL-2R+ tumors. In conclusion, the modification of NDV by the bispecific protein does not compromise severely the efficiency of gene delivery into IL-2R-positive tumors, but greatly reduces viral gene expression in IL-2R-negative tumors and in normal tissues.

Animals↗
PubMedDate not supplied

Newcastle disease virus passage in MDBK cells as an aid in detection of a virulent subpopulation.

Newcastle disease virus strains (NDV) La Sota, Texas GB (GB), and mixtures of the two strains were serially passaged in embryonated chicken eggs or the MDBK cell line, a more restrictive culture system than eggs for NDV. The two culture systems were compared by evaluating culture harvests for pathogenicity in inoculated chickens; the harvests were identified by hemagglutination-inhibition tests against monoclonal antibodies that can differentiate La Sota and GB cultures. Both viruses, inoculated alone or as mixtures, were propagated by passage in chicken eggs. La Sota strain failed to propagate by continuous passage in MDBK cells, and only GB was identified in culture harvests propagated in MDBK cells that had been inoculated with GB or mixtures of GB and La Sota. The results indicate that the MDBK cell line is a more selective substrate than chicken eggs and suggest that passage in MDBK cells may aid in selecting for more virulent subpopulations of NDV in a mixed culture. Other reference NDV strains, pigeon NDV isolates, and recent lentogenic NDV isolates from chickens were also passaged in MDBK cells; all strains except those that are classified as lentogens like La Sota could be serially propagated in MDBK cells.

Animals↗
PubMedDate not supplied

Protective immunity against Newcastle disease: the role of antibodies specific to Newcastle disease virus polypeptides.

Studies were performed to determine if passive immunization with hyperimmune sera generated to specific Newcastle disease virus (NDV) proteins conferred protection against virus challenge. Six groups of 3-wk-old chickens were passively immunized with antiserum against either hemagglutinin-neuraminidase/fusion, (HN/F) protein, nucleoprotein/phosphoprotein (NP/P), Matrix (M) protein, a mixture of all NDV proteins (ALL), intact ultraviolet-inactivated NDV (UVNDV), or negative sera. Blood samples were collected 2 days postimmunization, and the birds were challenged with Texas GB strain of NDV. Antibody titers were detected from those recipient birds that had received the antisera against the HN/F, ALL, or UVNDV by a hemagglutination inhibition test, an enzyme-linked immunosorbent assay (ELISA), and a virus neutralization test. Antibodies were detected only by the ELISA from the birds that had received antisera against NP/P and M protein. Antibody titers in the recipient birds dropped by two dilutions (log2) after 2 days postinjection. Birds passively immunized with antisera against HN/F, ALL, and UVNDV were protected from challenge, whereas chickens passively immunized with antisera against NP/P and M protein and specific-pathogen-free sera developed clinical signs of Newcastle disease. The challenge virus was recovered from the tracheas of all passively immunized groups. The presence of neutralizing antibodies to NDV provided protection from clinical disease but was unable to prevent virus shedding from the trachea.

Animals↗
PubMed2004

Nucleotide and predicted amino acid sequence analysis of the fusion protein and hemagglutinin-neuraminidase protein genes among Newcastle disease virus isolates. Phylogenetic relationships among the Paramyxovirinae based on attachment glycoprotein sequences.

Highly virulent Newcastle disease virus (NDV) isolates are List A pathogens for commercial poultry, and reports of their isolation among member nations must be made to the Office of International Epizootes (OIE). The virus is classified as a member of the order Mononegavirales in the family Paramyxoviridae of the subfamily Paramyxovirinae. Two interactive surface glycoproteins, the fusion (F) and hemagglutinin-neuraminidase (HN) proteins, play essential roles in NDV attachment and fusion of cells during infection. Antibodies to the F or HN proteins are capable of virus neutralization; however, no full-length sequences are available for these genes from recently obtained virulent isolates. Therefore, nucleotide and predicted amino acid sequences of the F and HN protein genes from 16 NDV isolates representing highly virulent viruses from worldwide sources were obtained for comparison to older virulent isolates and vaccine strains. The F protein amino acid sequence was relatively conserved among isolates maintaining potential glycosylation sites and C residues for disulfide bonds. A dibasic amino acid motif was present at the cleavage site among more virulent isolates, while the low virulence viruses did not have this sequence. However, a Eurasian collared dove virus had a K114Q substitution at the F cleavage site unique among NDV isolates. The HN protein among NDV isolates maintained predicted catalytic and active site residues necessary for neuraminidase activity and hemagglutination. Length of the HN for the Eurasian collared dove isolate and a previously reported heat resistant virulent isolate were longer relative to other more recent virulent isolates. Phylogenetically NDV isolates separated into four groups with more recent virulent isolates forming a diverse branch, while all the avian paramyxoviruses formed their own clade distinct from other members of the Paramyxoviridae.

Amino Acid Sequence↗
PubMed1995

Cell-mediated immunity in chickens vaccinated with the V4 strain of Newcastle disease virus.

A leukocyte migration inhibition assay was used to demonstrate antigen-specific cell-mediated immunity (CMI) in chickens vaccinated with the V4 strain of Newcastle disease virus. Chickens were vaccinated when 5 weeks old, and again 3 and 7 weeks later. CMI was detected 9 days after initial vaccination by eyedrop, but only after the second dose of vaccine delivered into the crop. In some chickens there was a temporary suppression of CMI to Newcastle disease virus, especially after the initial application of vaccine to the crop. Haemagglutination inhibition antibodies developed to similar levels in chickens after vaccination by either route. There was no obvious correlation between antibody and CMI responsiveness.

Animals↗
PubMed1977

[Immunogenic properties of 3 local lentogenic strains of the Newcastle disease virus used in the spray method of vaccination].

Three local lentogenic strains of the Newcastle disease virus, GT, D and R, were made use of to produce live liquid vaccines. These proved harmless and immunogenic when applied through the spray-method to broiler chicks, aged 5 days, at three successive revaccinations (on the 25th, 60th and 140th day of age) carried out by the same method. The strain D vaccine induced serum antihemagglutinins that had higher titers than those induced by vaccines of the other local strains and the standard strains Hitchner-B1 and La Sota.

Animals↗
PubMedDate not supplied

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

The virulence of six pigeon-origin isolates of Newcastle disease virus (NDV) was evaluated before and after passage in white leghorn chickens. Four isolates were defined as pigeon paramyxovirus-1 (PPMV-1) and two isolates were classified as avian paramyxovirus-1 (APMV-1) with NDV monoclonal antibodies. The four PPMV-1 isolates were passaged four times in chickens, and the APMV-1 isolates were passaged only once. Infected birds were monitored clinically and euthanatized. Tissues were collected for histopathology, in situ hybridization with a NDV matrix gene digoxigenin-labeled riboprobe, and immunohistochemistry with an anti-peptide antibody to the nucleoprotein. Mean death time, intracerebral pathogenicity index, and intravenous pathogenicity index tests performed before and after passage in chickens demonstrated increased virulence of the passaged PPMV-1 isolates and high virulence of the original isolates of APMV-1. Sequence analysis of the fusion protein cleavage site of all six isolates demonstrated a sequence typical of the virulent pathotype. Although the pathotyping results indicated a virulence increase of all passaged PPMV-1 isolates, clinical disease was limited to depression and some nervous signs in only some of the 4-wk-old specific-pathogen-free white leghorns inoculated intraconjunctivally. However, an increased frequency of clinical signs and some mortality occurred in 2 wk olds inoculated intraconjunctivally with passaged virus. Histologically, prominent lesions in heart and brain were observed in birds among all four groups inoculated with the PPMV-1 isolates. The behavior of the two pigeon-origin APMV-1 isolates when inoculated into chickens was characteristic of velogenic viscerotropic NDVs and included necro-hemorrhagic lesions in the gastrointestinal tract.

Amino Acid Sequence↗
PubMed2004

Interaction between genomes of infectious bronchitis and Newcastle disease viruses studied by reverse transcription-polymerase chain reaction.

Reverse transcription-polymerase chain reaction (RT-PCR) with specific primers for the S1 gene of IBV and for the fusion protein cleavage site of NDV was used for detection of Infectious bronchitis virus (IBV, the family Coronaviridae) and Newcastle disease virus (NDV) genomes. The sensitivity of IBV and NDV RT-PCR was 10(3.7) and 10(3.0) EID50, respectively. Although a multiplex RT-PCR could detect and differentiate NDV and IBV genomes present in the same sample, there was a slight inhibition of the IBV PCR if a high amount of NDV genome was present in the sample. To overcome this problem a separate PCR for each virus was used to assess the interaction between vaccine IBV and NDV either inoculated singly or together into chickens. In the group vaccinated with the Newcastle disease (ND) vaccine alone, the viral genome was detected on days 2, 4 and 7 post vaccination (p.v.), while in the chickens given the infectious bronchitis (IB) vaccine alone, the viral genome was detected only on day 4 p.v. In the group inoculated with both vaccine viruses there was a 10(3)-fold reduction in the cDNA dilution factor on day 4 p.v. for both IBV and NDV genomes. This demonstrated clearly that when both these vaccines are administered there is a transient reduction in the replication of both viruses, probably due to their competition for the same target epithelial cells in the respiratory tract.

Allantois↗
PubMed1990

Insertion of the fusion gene from Newcastle disease virus into a non-essential region in the terminal repeats of fowlpox virus and demonstration of protective immunity induced by the recombinant.

In this paper we report on the identification of non-essential genes in the terminal repeats of the avipox-virus fowlpox virus and the use of these as insertion sites in a vector system. Foreign genes inserted into these sites are shown to be present in two copies in the resultant recombinant virus. To test the potential use of this vector as a live vaccine the fusion gene of Newcastle disease virus has been inserted into a vaccine strain of fowlpox virus and inoculated into chickens. The experiments demonstrate the ability of the recombinant to protect chickens against challenge by a virulent strain of Newcastle disease virus and to elicit the formation of an anti-fusion protein antibody.

Animals↗
PubMed1969

Morphogenesis of Newcastle disease virus in chorioallantoic membrane.

Chick embryo chorioallantoic membrane, infected with the Blacksburg strain of Newcastle disease virus, was examined with an electron microscope to investigate the sequence of viral-induced host cell alterations. These were evident mostly in the endodermal epithelial cells lining the allantoic sac and were divided arbitrarily into three stages. Stage 1 was characterized by commencement of cell hypertrophy and hyperplasia and presence of fewer cytoplasmic inclusion bodies normally found in the cells; in stage 2, juxtanuclear nucleocapsid-glycogen aggregates appeared, and there were increased numbers of microvilli; stage 3 was characterized by increased cytoplasmic density and evidence of viral assembly and release. The morphological features of viral assembly and the virion are also described.

Animals↗
PubMedDate not supplied

Evidence of genetic heterogeneity of some lentogenic Newcastle disease virus strains.

Two distinguishable subpopulations were recovered from chickens infected with lentogenic strains of Newcastle disease virus originally ioslated from chickens in Australia and Ireland. In both instances, a subpopulation that rapidly eluted from chicken erythrocytes was recovered predominantly from tracheal swabs, and a slow-eluting subpopulation from cloacal swabs. The two subpopulations differed in thermostability of the hemagglutinatin and mean survival time of infected chicken embryos, as well as in elution pattern. The heterogeneity characteristic of the Aust-V4 strain and NI-Ulster strain could not be demonstrated in B1, NJ-LaSota, and ENG-F, lentogenic strains commonly used in vaccines. The question is discussed of whether standard vaccine strains are also hetrogeneous if a technique for separating stock populations is found, and the significance of possible heterogeneity to the effectivenss of the vaccines.

Agglutination Tests↗
PubMed1979

Molecular species of interferon induced in mouse L cells by Newcastle disease virus and polyriboinosinic-polyribocytidylic acid.

Interferons were stimulated in mouse L cells by Newcastle disease virus (NDV) or by polyriboinosinic-polyribocytidylic acid poly(rI).poly(rC). These were fractionated by sequential affinity chromatography on bovine plasma albumin (BPA)-Sepharose and on omega-carboxypentyl (CH)-Sepharose. Based on their interaction with CH-Sepharose, interferon induced by NDV was resolved into three major bands of activity (L/NDV-1,2,3) and poly(rI).poly(rC)-interferon into two (L/rI:rC-1,2). These interferon components were purified to a specific activity of 3 X 10(7) to 4 X 10(7) units/mg protein by antibody affinity chromatography and examined by electrophoresis in SDS-polyacrylamide gels. A total of five molecular species was thus identified for NDV-induced interferon and three for poly(rI).poly(rC) induced interferon, as summarized in Table 1. We conclude from our observations that mouse interferons can be produced by L cells in multiple forms with specific physiochemical properties and in proportions determined by the type of agent employed for induction.

Animals↗