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PubMedDate not supplied

Local pathological reactions and immune response of chickens to ISA-70 and other adjuvants containing Newcastle disease virus antigen.

Chickens were examined for gross and microscopic changes at the injection site for 16 weeks after an intramuscular injection of the following oil adjuvant emulsions containing inactivated Newcastle disease virus (NDV) antigen: ISA-70, Freund's incomplete adjuvant (FIA), Freund's complete adjuvant (FCA), and aluminum phosphate gel (APG). The reactions induced by ISA-70 emulsion were characterized by proliferation of macrophages, epithelioid cells, and fibroblasts around the small cysts in the muscle. Infiltration of lymphocytes and plasma cells also was present. These changes peaked 2 weeks postinjection and were milder than lesions due to the Freund's adjuvants. FCA induced the most severe granulomatous lesions with abscess formation. The reactions to APG were characterized by proliferation and accumulation of macrophages. The serological results revealed that the enhancing effect of ISA-70 on NDV antigen was the same as that of FIA.

Adjuvants, Immunologic↗
PubMedDate not supplied

Use of an avirulent Australian strain of Newcastle disease virus as a vaccine.

One-day-old chickens were transported from Australia to Malaysia and vaccinated orotracheally with an uninactivated vaccine prepared from avirulent Australian V4 strain of Newcastle disease virus (NDV). The vaccination regimes were as follows: group A, once, at 2 weeks old; group B, once, at 3 weeks old; group C, twice, at 2 and at 3 weeks old; group D, direct contact with groups A, B, and C; and group E, indirect contact with groups A, B, C, and D. Group F was unvaccinated controls. Challenge was with NDV virulent Ipoh AF 2240-226 strain, administered at 4 weeks old intramuscularly to 10 chickens in each group and orotracheally to 10 chickens in each group. The remaining chickens were challenged by contact with the inoculated chickens. Group mortalities following challenge were: A, 1/77; B, 1/34; C, 0/39; D, 0/45; E, 6/43; and F, 60/60.

Animals↗
PubMed1989

Proteins of Newcastle disease virus envelope: interaction between the outer hemagglutinin-neuraminidase glycoprotein and the inner non-glycosylated matrix protein.

Using linear sucrose-density ultracentrifugation analysis of Triton-solubilized Newcastle Disease Virus envelopes, we have evidenced, for the first time, the existence of interactions between the outer hemagglutinin-neuraminidase transmembrane glycoprotein and the inner non-glycosylated peripheral matrix protein. Such interactions seem to be electrostatic. These conclusions are based on the behavior of both proteins at different ionic strengths. When in low ionic strength buffer, hemagglutinin-neuraminidase and matrix proteins band together in the sucrose gradient, whereas at high ionic strength both proteins band at different rates in the gradient. The behavior of the inner matrix protein in our conditions was the expected one for a peripheral protein. The results of these 'in vitro' studies are also discussed in terms of the possible 'in vivo' role of such interactions.

Animals↗
PubMed1986

Anomalous behaviour of Newcastle disease virus haemagglutinin-neuraminidase protein in Western blotting analysis of monoclonal antibody binding sites.

Previous studies with a particular monoclonal antibody (MAb 445) raised against the Ulster strain of Newcastle disease virus (NDV) have shown that this MAb immunoprecipitates only the 74,000 mol. wt. (74K) haemagglutinin-neuraminidase protein (HN) of both Ulster and Beaudette C strains of NDV. Using the technique of Western blotting, it is shown that under certain denaturing conditions virion proteins comigrating in SDS-polyacrylamide gels with the faster migrating 67K fusion protein and 53K nucleocapsid/nucleocapsid-associated protein, as well as the 74K HN, all reacted strongly with MAb 445 and with MAb 14 which is also directed against the HN protein. Analysis of this anomalous behaviour using SDS-polyacrylamide diagonal gel electrophoresis has led to the unexpected conclusion that the 74K HN can electrophorese as three immunoreactive species of apparent mol. wt. 74K, 67K and 53K. If the Western blotting technique is to be applied to proteins which have not been sufficiently denatured (in an attempt to preserve epitope integrity) it is important to establish that no additional proteins remain associated with the protein bands that react with the MAb.

Antibodies, Monoclonal↗
PubMed1996

Newcastle disease viruses in birds in the Atlantic flyway: isolations, haemagglutination-inhibition and elution-inhibition antibody profiles.

The study involved 15 avian species with 5,012 attempts to isolate Newcastle disease viruses (NDV) from their faeces over a three-year period (1977-1979). NDV were isolated from asymptomatic adult Canada geese, nestling Royal terms, a juvenile European Mute swan, and adult Tundra swans on the Eastern flyway. Ring-billed gulls were negative for haemagglutination-inhibition (HI), elution-inhibition (EI) (anti-neuraminidase) antibodies, and NDV despite 3,403 isolation attempts. The EI antibody assay used a strain isolated from a Mute swan. The prevalence of EI antibodies in the swan and geese ranged from 3-41%, while the geometric mean titre (GMT) varied from 20-36. The prevalence of HI antibodies in the swans and geese ranged from 4-62%, while the GMT varied from 16-42. In each of three years (1977-1979), the adult Mute swans had a higher HI antibody prevalence than the juveniles (P < 0.01). Among the Mute swans the HI and EI assays detected serologic conversions and persistent antibodies over the three-year period. The HI and EI assays were effective in showing differences in antibody prevalences in populations of feral birds of different species and age. The EI assay is applicable for population studies of the anti-neuraminidase antibody.

Animals↗
PubMed1988

Nucleotide sequence of the fusion and haemagglutinin-neuraminidase glycoprotein genes of Newcastle disease virus, strain Ulster: molecular basis for variations in pathogenicity between strains.

The nucleotide sequences of the fusion (F) and haemagglutinin-neuraminidase (HN) glycoprotein genes of the extremely avirulent Newcastle disease virus (NDV) strain Ulster have been determined by sequencing cDNA clones derived from viral genomic RNA. Open reading frames, assumed to encode the F0 and HN0 glycoprotein precursors, were 553 and 616 amino acids long, respectively. Comparisons of the two glycoprotein sequences with those of more virulent NDV strains suggested an explanation for the molecular basis of the wide-ranging differences in virulence observed between strains of NDV. The open reading frame corresponding to the Ulster HN glycoprotein extended beyond the C terminus of more virulent strains. This C-terminal extension was assumed to be responsible for the origin of the HN precursor (HN0) found in strain Ulster and other extremely avirulent strains of NDV. There were fewer basic amino acids at the cleavage site of F0 in strain Ulster than are present in more virulent strains, which may be responsible for the absence of cleavage and activation of F0 from this strain in many host cells. In more virulent strains of NDV, as well as in other paramyxoviruses, a phenylalanine residue occurs at the N terminus of the F1 cleavage fragment. The occurrence of a leucine residue at this position in strain Ulster may be partly responsible for the lack of virulence of this strain.

Base Sequence↗
PubMed1984

The carboxyterminus of the hemagglutinin-neuraminidase of Newcastle disease virus is exposed at the surface of the viral envelope.

The amino-terminal and the carboxy-terminal amino acids of the hemagglutinin-neuraminidase glycoprotein of the Ulster strain of Newcastle disease virus have been analyzed before and after proteolytic activation of the precursor HNo (Mr approximately 82K). The amino termini of HNo and of the large cleavage fragment HN (approximately 74K) obtained by in vivo and in vitro proteolysis could not be sequenced by Edman degradation. This indicates that in both instances the amino termini are blocked. The carboxy termini of HNo and HN are different as demonstrated by end-point digestion with carboxypeptidase A. Furthermore, a small cleavage fragment (approximately 9K) of HNo that was removed from the virion after trypsin treatment could be purified by HPLC. In contrast to HN, this fragment displays a free amino terminus susceptible to Edman degradation. These data indicate that conversion of HNo involves removal of a 9K glycopeptide from the carboxy-terminal end. Thus, it has to be concluded that, unlike most other viral glycoproteins, the hemagglutinin-neuraminidase is inserted in the envelope with its carboxy terminus exposed at the surface of the virus particle.

Amino Acid Sequence↗
PubMed1994

Effective anti-metastatic melanoma vaccination with tumor cells transfected with MHC genes and/or infected with Newcastle disease virus (NDV).

The therapeutic efficacy of active immunization with B16-F10.9 melanoma cells transfected with syngeneic major histocompatibility complex (MHC) class-I genes, modified by infection with Newcastle Disease virus (NDV) or modified by both treatments, was compared. B16-F10.9 tumor-bearing mice were treated at various stages of tumor growth and metastasis with irradiated, modified tumor-cell vaccines. Irradiated tumor cells and H-2Db transfectants did not stimulate anti-tumor immunity while H-2Kb transfectants and NDV-modified F10.9 cells showing low and high expression of MHC class-I genes efficiently prevented metastasis of small established tumors. NDV-modified parental-cell vaccines functioned optimally and improved overall survival by about 60%, also at early stages of metastasis establishment. A synergistic effect of H-2Kb expression and virus modification on rejection of micrometastases was observed in mice bearing advanced tumors. Postoperative vaccination of mice carrying multiple metastases with NDV-modified vaccines caused significant, but incomplete, reduction of metastatic tumor load. The therapeutic effect of NDV-modified tumor vaccines was dependent on multiple immune mechanisms. Depletion of CD8, CD4 or NK cells by in vivo treatment with monoclonal antibodies reversed the immunotherapeutic effects of the vaccine. Thus, tumor xenogenization and gene modification may act synergistically to vaccinate against advanced tumors, while single modalities can effectively vaccinate against metastasis at early stages of tumor growth.

Animals↗
PubMed2004

Detection of Newcastle disease virus using a SYBR Green I real time polymerase chain reaction.

A two-step SYBR Green I real time polymerase chain reaction (PCR, real time PCR) for the detection of Newcastle disease virus (NDV) was developed. A melting curve analysis was performed to distinguish specific from non-specific products and primer dimers. Regardless of different virus pathotypes the melting temperature (Tm) ranged from 86 degrees C to 87 degrees C. The sensitivity of the real time PCR was compared with the reverse transcription (RT)-nested PCR enzyme-linked immunosorbent assay (ELISA, RT-nested PCR ELISA). Whereas the detection limit of the real time PCR was 10 pg DNA, the RT-nested PCR ELISA and conventional PCR could only detect up to 1 ng and 10 ng DNA, respectively. Thus the real time PCR offers a sensitive, rapid and convenient method for screening large number of NDV specimens.

Animals↗
PubMed1989

Functional and neutralization profile of seven overlapping antigenic sites on the HN glycoprotein of Newcastle disease virus: monoclonal antibodies to some sites prevent viral attachment.

We have previously identified five antigenic sites on the hemagglutinin-neuraminidase (HN) glycoprotein of the Australia-Victoria isolate of Newcastle disease virus (Iorio and Bratt, J. Virol. 48, 440-450; Iorio et al., J. Gen. Virol. 67, 1393-1403). Two additional sites (designated 12 and 23) are now described, bringing to a total of seven the number of antigenic sites defined by our panel of neutralizing anti-HN antibodies. Competition antibody binding and additive neutralization assays reveal that each of these newly-identified sites overlaps two previously-defined ones. The seven HN antigenic sites thus form a continuum in the three-dimensional conformation of the molecule. Studies on the inhibition of hemagglutination (HA), neuraminidase (NA) and the attachment of virus to chick cell monolayers have been used to construct a functional profile of each antigenic site. Monoclonal antibodies (mAbs) to three overlapping sites (12, 2 and 23) inhibit HA and NA and prevent viral attachment to chick cell monolayers. These findings are consistent with the domains recognized by these mAbs being close to the NA and receptor-binding sites. MAbs to two other overlapping sites, 14 and 1 (which in turn, overlap site 12), inhibit HA quite effectively, and attachment to a lesser extent. Sites 14 and 1 probably identify a second domain involved in receptor recognition. MAbs to the two remaining sites (3 and 4), though neutralizing, are negative in all three assays, thus recognizing domains not involved in HA or NA or attachment to chick cells.

Animals↗
PubMed1980

Mutational changes of the protease susceptibility of glycoprotein F of Newcastle disease virus: effects on pathogenicity.

Two chemically induced mutants with an alteration in the susceptibility of glycoprotein F to proteolytic cleavage have been isolated from the apathogenic strains of La Sota and Ulster of Newcastle disease virus. In contrast to the La Sota wild type, cleavage of the precursor F0 and activation of cell fusing activity and infectivity take place if the mutant is grown in MDBK and BHK21-F cells. The mutant is, therefore, able to undergo multiple replication cycles in cells non-permissive for the wild type. This increase in host range is paralleled by an increase in pathogenicity for chick embryos. The increase in host range of the Ulster mutant is less distinct. This mutant, which does not differ in pathogenicity from its wild type, produces in MDBK cells incompletely activated virus containing predominantly glycoprotein HN in the uncleaved and glycoprotein F in the cleaved form. The data support the concept that the susceptibility of the virus glycoproteins to proteolytic activation is an important factor in determining the pathogenicity of this virus.

Animals↗
PubMed2004

RNA-dependent RNA polymerase gene analysis of worldwide Newcastle disease virus isolates representing different virulence types and their phylogenetic relationship with other members of the paramyxoviridae.

Nucleotide sequence was determined for the RNA-dependent RNA polymerase (L) gene of 16 Newcastle disease virus (NDV) isolates from diverse geographic and chronological origins. The observed consensus amino acid sequence conformed to the six domains previously identified among paramyxovirus L proteins, and the putative 749QGDNQ753 active site was strictly conserved among all isolates. Analysis of predicted amino acid sequences allowed us to identify a sequencing error in the previously reported L genes for NDV. The correct sequences reported herein provided a more accurate alignment with predicted l-amino acid sequences of other paramyxoviruses. Comparison of L gene coding sequences among isolates revealed that synonymous substitutions dominated non-synonymous substitutions, as observed previously with other NDV genes. However, the overall substitution rate was lower than other genes examined making the L gene the most conserved of the NDV genome. Phylogenetic analysis of L genes among NDV isolates was consistent with previous results that suggested the existence of two major lineages. One group contained strains isolated in North America prior to 1970 and included virulent and vaccine strains, while the second group included virulent viruses isolated worldwide. A comparison of the NDV L coding sequences to other Paramyxoviridae illustrated the unique clustering of the avian-specific paramyxoviruses, further justifying the newly created Avulavirus genus.

Animals↗
PubMedDate not supplied

Experimental Newcastle disease virus infections in three species of wild birds.

Three species of wild birds--the red-winged blackbird (Agelaius phoeniceus), an African weaver finch (Quelea quelea), and the sandhill crane (Grus canadensis)--were exposed to Newcastle disease virus (NDV) to determine susceptibility and host response. The NDV used were a vaccine strain (LaSota) and a viscerotropic velogenic strain (New York Parrot). Host response was monitored by virus isolation, signs of disease, and serologic response. Both the red-winged blackbirds and the quelea shed little virus and developed low antibody titers. A low mortality in the blackbirds did not appear to be due to ND alone. Vaccinated cranes were well protected against velogenic challenge, whereas unvaccinated cranes shed the velogenic virus from the cloaca for weeks. The ramifications of the low antibody titers produced in birds of two species are discussed, as well as the potential involvement of each species in transmission of the virus.

Animal Population Groups↗