Infection of the upper respiratory tract of the chick with a mild (vaccine) strain of Newcastle disease virus. II. Studies on the pathogenesis of the infection.
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Animals↗
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NUTRITION AND BIOCHEMISTRY OF SURVIVAL DURING NEWCASTLE DISEASE VIRUS INFECTION. III. RELATION OF DIETARY PROTEIN TO NUCLEIC AND FREE AMINO ACIDS OF AVIAN LIVER.
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Alanine↗
Comparison of dimethyl dioctadecyl ammonium bromide, Freund's complete adjuvant and mineral oil for induction of humoral antibodies, cellular immunity and resistance to Newcastle disease virus in chickens.
Dimethyl dioctadecyl ammonium bromide (DDA), a lipophilic quaternary amine, was evaluated in adult chickens for potentiation of immunological responses to subcutaneously administered inactivated Newcastle disease virus (NDV) vaccines. DDA enhanced humoral and cell-mediated immune (CMI) responses to levels which were significantly higher than those induced by the vaccine alone The haemagglutination inhibition antibody titers induced by DDA were slightly lower than those induced by mineral oil although neutralizing antibody titers seemed to be higher. DDA induced strong CMI (DTH and lymphocyte proliferation) responses, more than those induced by Freund's complete adjuvant and mineral oil. Both DDA and mineral oil induced comparable high levels of protection to challenge doses of 200,000 LD50 per chicken. No toxic effects or local tissue damage were observed in any of the inoculated chickens.
Adjuvants, Immunologic↗
[Virus-specific poli(A)-containing RNA-transcripts in cells infected with Newcastle disease virus].
The techniques of chromatography on poly (U)-cellulose and sedimentation and electrophoresis analysis were used to study poly(A)-containing virus-specific RNA of Newcastle disease virus. The poly(A)-containing molecules were shown to be present in approximately similar amounts (60-80%) in 18S, 24S, and 35S RNA. No poly(A)-containing molecules capable of interaction with poly(U) sepharose were found in 50S RNA. In 18S RNA there are 6 individual poly(A)-containing RNA-transcripts with molecular weights well correlating with those of virus proteins. The average size of poly(A) for 18S and 24S RNA is 60-70 nucleotide residues. In the sedimentation analysis of poly; A)-containing 24S and 35S RNA eluted from poly(U)-columns, some of the material sedimented in the zone of 18S RNA. The presence of complexes of two or several molecules with the sedimentation coefficient 18S in these RNA's is suggested.
Base Sequence↗
[Antibody reaction to inhalation vaccination of poultry against Newcastle disease].
Aerosol preparated from the AVIPEST vaccine by means of three types of aerosol generators was used for vaccination of 294 chickens of different age against the Newcastle disease in laboratory tests and 30 000 chickens at the age of two and three weeks under field conditions. The general and local antibody reactions to vaccination were evaluated on the basis of the determination of the content of hemagglutination-inhibition and neutralization antibodies in the blood serum and in respiratory-tract flushings. The level of secretory antibodies in the flushings from the respiratory tract, determined by the described original method, is an important parameter of the resistance level of the organism to infection. The highest average levels of antibody titres in serum and in flushings were obtained 14 to 21 days from vaccination (7-10 log2). The La Sota vaccination strain was eliminated from the organism after spray vaccination and immunized the non-vaccinated contact chicken. Revaccination increased and greatly prolonged immunity. All the chickens inoculated showed resistance to challenge infection whereas the control birds died within four to six days.
Aerosols↗
Multiplication of the V4 strain of Newcastle disease virus in Madin Derby bovine kidney cells.
This study describes a reproducible cell culture system that permits the growth and secondary multiplication of the V4 strain of Newcastle disease virus. Allantoic fluid, magnesium chloride and diethylaminoethyl dextran were incorporated in Dulbecco's modified Eagle's medium to encourage secondary viral multiplication without adversely affecting healthy Madin Derby bovine kidney cell growth.
Allantoin↗
Nucleotide sequence of the gene encoding the Newcastle disease virus hemagglutinin-neuraminidase protein and comparisons of paramyxovirus hemagglutinin-neuraminidase protein sequences.
The nucleotide sequence of cloned cDNA copies of the mRNA encoding the Newcastle disease virus (NDV), strain A-V, hemagglutinin-neuraminidase (HN) protein was determined. A single open reading frame in the sequence encodes a protein of 570 amino acids with a calculated molecular weight of 62,280. The predicted protein sequence contains only one obvious potential membrane spanning region, located 27 amino acids from the amino terminus of the sequence. The predicted sequence contains 6 glycosylation sites and 14 cysteine residues. Comparison of the NDV HN protein sequence with three other paramyxovirus HN protein sequences reveals two regions that have homologies in all four sequences. The conserved cysteine residues are clustered in these two regions. One conserved region is located near the middle of the predicted sequence while the second region is in the carboxy terminal third of the molecule. The presence of conserved regions suggests the importance of these areas of the molecule in the structure or function of the protein.
Amino Acid Sequence↗
A temperature-sensitive mutant of Newcastle disease virus which is affected in both haemagglutinin-neuraminidase and matrix proteins.
Virions prepared from a non-revertible temperature-sensitive (ts) mutant (ts53) of Newcastle disease virus (NDV) grown in ovo at the permissive temperature (34 degrees C) possessed thermolabile haemagglutination and neuraminidase activities compared with parental (ts+) virions. Purified haemagglutinin-neuraminidase (HN) protein from ts53 virions was also more thermolabile than ts+ HN protein. SDS-PAGE analysis of [3H]leucine pulse- and pulse/chase-labelled NDV proteins synthesized in chick embryo fibroblasts following infection with ts+ and ts53 virus revealed that ts53 matrix (M) protein was unstable and disappeared during chase incubations only at the non-permissive temperature (42 degrees C). The non-revertibility of the ts53 mutant may indicate that it is a double mutant affected in both HN and M genes; alternatively this mutant may only be affected in the HN gene, the close physical association of the thermolabile HN with the M protein during virus maturation resulting in the lack of protection of the M protein from the action of cellular proteases at the non-permissive temperature.
Cycloheximide↗
NUTRITION AND BIOCHEMISTRY OF SURVIVAL DURING NEWCASTLE DISEASE VIRUS INFECTION. II. RELATION OF CLINICAL SYMPTOMS AND STARVATION TO NUCLEIC AND FREE AMINO ACIDS OF AVIAN LIVER.
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Alanine↗
Newcastle disease virus-specific RNA: hybridization-competition of the non-dissociable 35 S RNA with individual 18 S RNA species.
The non-dissociable fraction of Newcastle disease virus- (NDV-Y specific 35 S RNA competed efficiently with two individual 18 S RNA transcripts in hybridization-competition experiments with 50 S virion RNA. The third 18 S component could not be displaced from the hybrid by an excess of non-dissociable 35 S RNA.
Base Sequence↗
Mature, cell-associated HN protein of Newcastle disease virus exists in two forms differentiated by posttranslational modifications.
Characterization of the posttranslational modifications of the mature, cell-associated hemagglutinin-neuraminidase (HN) protein of Newcastle disease virus (NDV) revealed that the HN protein exists in two forms differentiated by disulfide bonds and glycosylation. One form, HNa, contains intermolecular disulfide bonds and is endoglycosidase H partially resistant. The other form, HNb, is not linked by disulfide bonds and is endoglycosidase H sensitive. Both forms of the protein are modified with fucose indicating transport to the Golgi membranes. Both forms are detected at the cell surface by monoclonal antibody. Furthermore, both forms are transported to the cell surface with identical kinetics. HNa is incorporated into virions. HNb is not incorporated into virions and is presumably degraded. The cDNA derived from the HN gene was expressed from a retrovirus vector. The majority of the protein expressed was in the nonvirion-associated form b. Evidence is presented that the level of gene expression determines the ratio of the two forms of HN protein. At high levels of expression, the virion-associated form is favored while at low levels of expression the nonvirion-associated form is favored. The results presented have implications for persistent infections as well as expression of viral genes from different vectors.
Animals↗
Induction of interferon-alpha and tumor necrosis factor-related apoptosis-inducing ligand in human blood mononuclear cells by hemagglutinin-neuraminidase but not F protein of Newcastle disease virus.
To determine molecular viral components which can induce innate immune responses in human peripheral blood mononuclear cells (PBMC), we investigated the anti-neoplastic agent Newcastle disease virus (NDV) and its two spike proteins hemagglutinin-neuraminidase (HN) and fusion protein (F). NDV was an excellent inducer in PBMC of IFN-alpha production and capable of inducing upregulation of plasma membrane expression of tumor necrosis factor related apoptosis inducing ligand (TRAIL). Viral replication was not required for these responses because NDV inactivated for 5 min by UV was as good as live NDV. NDV-modified and paraformaldehyde-fixed BHK cells could also trigger IFN-alpha and TRAIL induction, indicating that contacts of responder cells with NDV-modified cell surfaces are sufficient to induce these activities in PBMC. Antibodies against HN but not F were able to block these responses. Finally we could show that HN but not F induced IFN-alpha and TRAIL in PBMC. This was possible through the use of respective gene transfectants generated with the help of Semliki Forest virus (SFV) replicase-based DNA recombinant expression systems. Upon contact with BHK cells expressing HN but not F at their cell surface, human PBMC produced IFN-alpha and some cells, including monocytes and T lymphocytes, upregulated cell surface TRAIL expression.
Apoptosis Regulatory Proteins↗
[Effect of 5-iodo-2-deoxyuridine (IDU) on Newcastle disease virus replication in cell cultures].
In chick embryo fibroblasts and in mouse fibroblasts of established L line, grown in the presence of IDU, the multiplication of Newcastle Disease virus was better and the CPE more distinct than in control cell cultures. In calf kidney cells only the CPE was enhanced. The most pronounced influence of IDU on virus multiplication was found after adding the analogue in maximum tolerated dose to the medium at the moment of cell culture preparation. The growth curve of virus in chick embryo fibroblasts grown in the presence of IDU shows the most distinct influence of this analogue with regard to LaSota strain. Already two days after inoculation with a smaller dose the titer was 2 log higher than in control cultures and the maximum difference of 3 log was found on the 5th day. In reference to Roakin strain the influence of IDU was weaker--maximum difference of 1,2 log on the fifth day after inoculation. Also the difference between the titers of virus in L cells grown in the presence of IDU and in the control ones, inoculated with LaSota strain was greater (about 1,5 lg during 2-8 days) than in the case of Roakin strain. In chick embryos treated with IDU intraallantoically at 24 and 48 hours before the inoculation with LaSota strain (only this was used) the enhancement of virus multiplication was much poorer than in chick embryo fibroblasts.
Animals↗
The human pathogenicity of the Newcastle disease virus and its relation to the mumps, fowl plague and influenza viruses.
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Animals↗
Structural analysis of a designed inhibitor complexed with the hemagglutinin-neuraminidase of Newcastle disease virus.
Viruses of the Paramyxoviridae family are the leading cause of respiratory disease in children. The human parainfluenza viruses (hPIV) are members of the Paramyxovirinae subfamily, which also includes mumps virus, Newcastle disease virus (NDV), Sendai virus (SV) and simian type 5 virus (SV5). On the surface of these viruses is the glycoprotein hemagglutinin-neuraminidase (HN), which is responsible for cell attachment, promotion of fusion and release of progeny virions. This multifunctional nature of HN makes it an attractive target for the development of inhibitors as a treatment for childhood respiratory diseases. Here we report the crystal structure of NDV HN in complex with a derivative of 2-deoxy-2,3-dehydro-N-acetylneuraminic acid, Neu5Ac2en, that has a functional group designed to occupy a large conserved binding pocket around the active site. The purpose of this study was to examine the effect of a bulky hydrophobic group at the O4 position of Neu5Ac2en, given the hydrophobic nature of the binding pocket. This derivative, with a benzyl group added to the O4 position of Neu5Ac2en, has an IC(50) of approximately 10 microM in a neuraminidase assay against hPIV3 HN. The IC(50) value of the parent compound, Neu5Ac2en, in the same assay is approximately 25 microM. These results highlight the striking difference between the influenza neuraminidase and paramyxovirus HN active sites, and provide a platform for the development of improved HN inhibitors.
Crystallography, X-Ray↗
Identification and grouping of Newcastle disease virus strains by restriction site analysis of a region from the F gene.
A 75% region of the F gene (between nucleotides 334 and 1682) of Newcastle disease virus (NDV) RNA was amplified by reverse transcription polymerase chain reaction (RT-PCR). PCR products were cleaved by three restriction endonucleases and the positions of thirty cleavage sites were mapped in more than 200 NDV strains. Restrictions site analysis established six major groups of NDV isolates and unique fingerprints of vaccine strains. Group I comprised lentogenic strains isolated mainly from waterfowl with some from chickens. "Old" (prior to 1960s) North American isolates of varying virulence including lentogenic and mesogenic vaccine strains belonged to group II. Group III included two early isolates from the Far East. Early European strains (Herts 33 and Italien) of the first panzootic (starting in the late 1920s) and their descendants with some modifications were placed into group IV. NDV strains isolated during the second panzootic of chickens (starting in the early 1960s) were classified into two groups. Group V included strains originating in imported psittacines and in epizootics of chickens in the early 1970s. Group V1 comprised strains from the Middle East in the late 1960s and later isolates from Asia and Europe. Pigeon paramyxovirus-1 strains that were responsible for the third panzootic formed a distinct subgroup in group V1. Our grouping of NDV strains has confirmed group differences established by monoclonal antibodies. It is concluded that restriction site analysis of F gene PCR amplicons is a relatively fast, simple and reliable method for the differentiation and identification of NDV strains.
Animals↗
Evaluation of the efficacy of the crude extracts of Capsicum frutescens, Citrus limon and Opuntia vulgaris against Newcastle disease in domestic fowl in Tanzania.
Prophylactic and therapeutic efficacy of a combination of Capsicum frutescens (red pepper), Citrus limon (lemon) and Opuntia vulgaris (prickly pear) against Newcastle disease (ND) in domestic fowl were evaluated. Eighty-eight broiler chickens were divided into five groups. Birds from three groups were inoculated with velogenic ND virus strain, whereas birds from two groups were left as controls. Two groups received a mixture of the plant extract three days prior to inoculation and birds from one group were given the plant extract for two days following development of clinical signs. Blood samples were collected for haemaglutination inhibition tests (HI) for detection of ND virus antibodies. Body weights were monitored during the experiment. Three birds died from the group that was inoculated with ND virus and treated with the plant extract; two died from the group that received the plant extract as a prophylaxis and inoculated with ND virus; and one bird died from the group that was inoculated with ND virus but not given the plant extract. No death was observed in any of the birds in the control groups. Antibody titers for ND virus rose four-fold in the inoculated birds but remained low in the un-inoculated groups. Mean body weights of birds in group B declined markedly compared to the other groups. The results indicated that there was no prophylactic or therapeutic value of the plant extract against ND. The plant extract showed a negative effect on body weights in birds with ND.
Animals↗
Interferon-induced expression of If-1h and If-1l alleles in Newcastle disease virus-infected mouse macrophages is associated with specific differences in viral gene transcription.
We have studied the expression of cytokines and viral genes induced by Newcastle disease virus (NDV) and Sendai virus in bone marrow-derived macrophages (BMM) and lymphocytes from C57BL/6 mice and the congenic line B6.C-H-28c. These mice carry the loci If-1h (high) or If-1l (low), respectively, that are responsible for up to tenfold differences in the interferon (IFN)-alpha, IFN-beta, interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-alpha) response to NDV but not to Sendai virus. Only BMM but not spleen lymphocytes showed allele-specific differences in NDV-induced cytokine levels, indicating cell-specific If-1 expression. The If-1 locus harbors IFN-inducible gene(s) whose expression is prevented in the presence of cycloheximide. Our data provide evidence that the If-1l allele acts by specifically suppressing the cytokine response to NDV. Cytokine production was dependent on infectious virions, and kinetic analyses revealed a close correlation between the amount of viral transcripts and individual cytokine mRNA. BMM from lf-1l mice strongly restricted transcription of the NDV nucleoprotein (NP) gene, whereas BMM from If-1h mice supported NP transcription. Following treatment with IL-4, which inhibited constitutive IFN-beta gene expression, however, If-1l BMM became highly permissive for transcription of the viral NP gene and released high amounts of cytokines. We conclude that If-1l gene products are responsible for the low producer phenotype by efficiently interfering with NDV transcription, leading to strongly reduced intracellular levels of cytokine inducing viral dsRNA intermediates.
Alleles↗