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PubMed2000

Crystallization of Newcastle disease virus hemagglutinin-neuraminidase glycoprotein.

The hemagglutinin-neuraminidase (HN) glycoprotein of Newcastle disease virus was isolated by cleaving HN (cHN) from reconstituted virosome with chymotrypsin. N-terminal sequence analysis of the purified cHN showed that chymotrypsin cleavage had occurred at amino acid 123, freeing the C-terminal 454 amino acids. The purified cHN retained its neuraminidase and receptor binding activities and reacted with specific monoclonal antibodies, showing that the isolated cHN was biologically and antigenically functional. The crystals of the cHN were obtained in acetate buffer (pH 4.6) containing polyethylene glycol 3350 and ammonium sulfate and belong to the orthorhombic space group P2(1)2(1)2(1) with unit cell dimension of approximately a = 72 A, b = 78 A, and c = 198 A. Crystals of cHN grown in the presence of sialic acid (Neu5Ac) were grown in HEPES buffer (pH 6.2) containing polyethylene glycol 3350 and belong to the hexagonal space groups P6(1) or P6(5) with unit cell dimensions of a = b = 137.5 A and c = 116.6A. The orthorhombic crystals produced in this study diffract X rays to at least 2.0-A resolution, thereby setting the stage for the solution of the three-dimensional structure of the HN glycoprotein of a paramyxovirus.

Amino Acid Sequence↗
PubMedDate not supplied

Tween 80-solubilized Newcastle disease virus prepared as a water-in-oil-in-water vaccine.

The Newcastle disease virus (NDV) was solubilized with 10% w/v Tween 80 and inactivated with 0.05% v/v formalin. The average molecular mass of the released antigenic subunits was 307 kD. The tested vaccine was prepared in the form of a water-in-oil-in-water emulsion (WOWE) vaccine. The oil-to-aqueous ratio was 1:2. The solubilized NDV was administered alone or built in a tetravalent WOWE vaccine. A dose of the monovalent vaccine containing an equivalent of 44.7 microliters of detergent-treated NDV-allantoic fluid (NDV-AF) was sufficient for the complete protection of the commercially available chickens vaccinated at the age of 5 wk and challenged 7 wk later. The anti-NDV-free chickens, vaccinated at 4 wk of age and challenged 2 wk postvaccination, were 100% and 73% protected by a vaccinal dose containing 178.6 and 89.3 microliters of detergent-treated NDV-AF, respectively. Commercially available light pullets, primary vaccinated with live lentogenic NDV vaccine, generated a protective level of NDV antibodies after revaccination with WOWE vaccine containing 89.3 microliters of detergent treated NDV-AF. Laying hens were revaccinated under field conditions at the beginning of the laying cycle by the tetravalent vaccine. A vaccinal dose/bird containing 11.2 microliters of detergent-treated NDV-AF elicited a long-lasting high level of NDV neutralizing antibodies.

Animals↗
PubMed2000

Characterization of Newcastle disease viruses isolated from chickens and ducks in Tamilnadu, India.

During 1993, outbreaks of Newcastle disease occurred on many farms in Tamilnadu, India. Six Newcastle disease virus (NDV) isolates were obtained from the chickens on five different farms and from the birds on one duck farm during outbreaks of the disease. All the isolates were characterized as velogenic, based on the mean death time, intravenous pathogenicity index, intracerebral pathogenicity index (ICPI), stability of haemagglutinin at 56 degrees C, agglutination of equine erythrocytes, haemagglutination elution pattern and adsorption of haemagglutinin by chick brain cells. The isolate obtained from ducks resembled a group D strain, based on its ICPI and its reaction with a panel of monoclonal antibodies. The other five NDV isolates obtained from chickens were placed in groups B(1), C1(2) and D(2) on the basis of their binding patterns with the panel of monoclonal antibodies. In challenge experiments, it was found that LaSota vaccine provided 100% protection against each of these field isolates and against a local NDV strain obtained from the Institute of Veterinary Preventive Medicine, Tamilnadu, India, while unvaccinated chickens succumbed to challenge. The possible origin of epizootic viruses causing outbreaks in vaccinated flocks is discussed.

Animals↗
PubMedDate not supplied

Attenuation of lentogenic Newcastle disease virus strain B-1 by cold adaptation.

The Hitchner B-1 strain of Newcastle disease virus was plaque-cloned and then serially passaged 36 times in specific-pathogen-free (SPF) chicken embryos incubated at two different temperatures. Virus passaged at a reduced temperature (29 C) was identified as cold-adapted (Ca) and virus passaged at the normal temperature (37 C) was designated non-cold-adapted (non-Ca). The Ca and non-Ca B-1 viruses were compared with the parent B-1 and a commercial B-1 vaccine. In vitro Ca B-1 characteristics included adaptation for more rapid growth at 29 C and the aquisition of temperature sensitivity indicated by substantially reduced growth at 41 C, properties not seen with non-Ca B-1. Embryo mean death times for the Ca virus (140 hr) were longer than for non-Ca B-1 (107 hr) and parent B-1 (121 hr) viruses. The Ca virus retained a rapid (< 2 hr) hemagglutination (HA) elution rate but lost the property of binding the monoclonal antibody AVS-I typical of other B-1 strains. The pathogenicity of the Ca B-1 strain was compared to the non-Ca B-1, parent B-1 strain, and a commercial B-1 strain vaccine in 1-day-old broiler-type chickens. Pathogenicity was evaluated by assessing the severity of respiratory disease signs and the incidence of airsacculitis, perihepatitis, and pericarditis lesions in inoculated chicks. A respiratory disease index was calculated for each B-1 strain based on daily observation scores that determined the presence or absence of disease signs (coughing, rales, labored breathing, death) from 1 to 14 days following intratracheal inoculation with 10(6) 50% egg infective doses of virus per chick. The lower respiratory disease index obtained for the Ca B-1 strain (0.075) indicated it was less pathogenic than the commercial B-1 vaccine (0.296) and the non-Ca (0.478) and parent (0.521) B-1 strains. Ca B-1-infected chicks had only a 5% incidence of air sac lesions, compared to chicks given non-Ca (65%), Hitchner B-1 (65%), or a commercial B-1 vaccine (30%). Immunogenicity tests performed in 1-week-old SPF leghorn chickens demonstrated that Ca B-1 induced complete protection when administered intraocularly as a single entity. However, when Ca B-1 was given in combination with a modified live infectious bronchitis virus vaccine, chickens were only partially protected (60-75%) against Texas GB strain-induced neurotropic velogenic Newcastle disease.

Animals↗
PubMed1987

Suppression of contact sensitivity by a plastic adherent T-cell, induced in mice infected with Newcastle disease virus (NDV).

Previous studies have demonstrated that lymph node cells of mice skin-sensitized 24 h before are able to transfer contact sensitivity (CS) in naive recipients. These antigen presenting cells (APC) lose the ability to induce CS when donor mice are treated with the virus of Newcastle Disease (NDV) at the time of sensitization. In this paper we demonstrate that the cell capable of suppressing CS is a virus induced plastic adherent T-cell, which inhibits otherwise normal APC. In fact, the APC in infected mice are fully competent, as demonstrated by their ability to transfer CS, if the adherent T-cell population is removed by plastic adherence. Analysis shows that the CS suppressing adherent T-cells are Thy 1.2+, Lyt 1.1+ and I-J+ subset. The inhibition of CS by the NDV induced adherent T-cell is antigen non-specific and genetically restricted. We have also demonstrated that picrylated cells from NDV infected mice fail to trigger the release of non-specific inhibitor (nsINH) in the T-suppressor circuit. The effect of the adherent suppressor T-cell in that circuit was determined and the results indicate that the virus induced T-cell is able to suppress the release of nsINH by blocking the function of the APC.

Animals↗
PubMed1984

The effect of protein malnutrition on the susceptibility of the chicken nose to Newcastle disease virus.

The effect of protein malnutrition on the susceptibility of the chicken nose to Newcastle disease virus (NDV) was studied. Chicks that were rendered acutely protein-deficient for 1 or 3 weeks from the time of hatching yielded approximately 10 times more NDV from their throats than did normally fed animals 7 days after inoculation with the virus. Serum antibody responses in the protein-deficient animals 14 days after inoculation of NDV was statistically lower than in the normally fed animals. These results were supported by the mucociliary transport time in the turbinate and sinus, as well as by the histological changes in the turbinate. This difference may thus be due to the fact that normally fed maturing chicks develop an increased resistance to virus which is depressed in protein-deficient animals.

Animals↗
PubMed1982

Detection and quantitation of Newcastle disease virus proteins in infected chicken embryo cells.

A technique was analyzed by which Newcastle disease virus (NDV) proteins could be quantitatively detected in the presence of chicken embryo cellular proteins in NDV-infected cells. The technique involved removal of electropho-proteins from a sodium dodecyl sulfate-polyacrylamide-agarose gel matrix by chemical cleavage of the acrylamide gel cross-linker. The proteins were subsequently transferred and covalently bound to diazobenzyloxymethyl paper. By incubating the paper with unlabeled antisera and 125I-labeled Staphylococcus aureus protein A, the specificity of the antisera and the sensitivity of this method of quantitative antigen detection were tested. The results demonstrated that as little as 1 ng of an individual NDV protein could be detected. Furthermore, this technique can simultaneously quantitate the synthesis of multiple NDV proteins under experimental conditions in which immunofluorescence, hemadsorption, and plaque assays failed to show virus protein synthesis or the formation of virus progeny.

Animals↗
PubMed1981

Identification of the P proteins and other disulfide-linked and phosphorylated proteins of Newcastle disease virus.

A unique abundant protein, designated P by analogy to the putative polymerase proteins of other paramyxoviruses, was identified in purified Newcastle disease virus. Under nonreducing conditions the P proteins could be separated from other viral proteins on sodium dodecyl sulfate-polyacrylamide gels. The P proteins were isolated from detergent-solubilized virions as 53,000- to 55,000-dalton monomers and disulfide-linked trimers. Distinct forms of P having four different isoelectric points and two different electrophoretic mobilities were resolved by two-dimensional electrophoresis. Two forms of P were phosphorylated, as were the nucleocapsid protein and non-glycosylated membrane protein. In addition to disulfide-linked forms of P, dimers of the hemagglutinin-neuraminidase glycoprotein and two disulfide-linked versions of the fusion glycoprotein were identified. Several electrophoretic variants of the nucleocapsid protein that were probably created by intrachain disulfide bonding were also isolated from virions under nonreducing conditions. The locations of the newly identified proteins were determined by detergent-salt fractionation of virions and by surface-selective radioiodination of the viral envelope. The P proteins were associated with nucleocapsids and were not detected at the surface of virions. Both forms of the fusion glycoproteins were on the exterior of the viral envelope. Herein the properties of the P proteins are compared with similar proteins of rhabdoviruses and other paramyxoviruses, and a role for multiple forms of proteins in the genetic economy of newcastle disease virus is discussed.

Disulfides↗
PubMed1983

Freedom of coccidial oocysts from Newcastle disease virus.

Oocysts were recovered from chickens experimentally infected with the Mukteswar strain of Newcastle disease virus and Eimeria acervulina or E tenella. Oocysts sterilised by sodium hypochlorite solution were washed, ruptured and inoculated into embryonating eggs which were examined after five days by the haemagglutination test for virus. All these tests were negative. Virus, initially detected with oocysts separated from faeces, was no longer present after sporulation in 2 per cent potassium bichromate. The B1 strain of virus survived for up to 30 hours in 2 per cent sodium hydroxide and 2 per cent potassium bichromate but was quickly destroyed by sodium hypochlorite (specific gravity 1.075).

Animals↗
PubMed1978

Cross-linking of Newcastle disease virus (NDV) proteins.

The proxomity and spatial relationships of the structural proteins of Newcastle disease virus (NDV) were studied by chemical cross-linking with a series of imidoesters. When the virions were reacted by the cross-linker with a distance 6.1A or longer between the functional groups and analyzed by polyacrylamide gel electrophoresis, remarkable changes were observed in the migration patterns of the viral proteins. The most striking one was the extensive decrease in the intensity of the M protein band, and although not so strikingly, glycoprotein and nucleocapsid protein bands were reduced significantly. Instead, several protein complexes appeared at and near the top of the gels. The protein complexes formed by a reversible cross-linker, dimethyl-3,3'-dithiobispropionimidate (DTBP), were analyzed by two dimensional electrophoresis; the complexes on the first-dimension cylindrical gels were cleaved by reduction with 2-mercaptoethanol and electrophoresed laterally on the second-dimension slab gels. The results indicated that homodimers of glycoprotein, nucleocapsid protein and M protein were generated under the condition of the most gentle cross-linking employed. At the same time, however, trimer and higher homopolymers of M protein were already detectable. Under the more extensive conditions, the bulk of M protein was cross-linked to form a large protein complex with very high molecular weight. Further, small but significant amounts of glycoprotein and nucleocapsid protein were always detected in this complex. These results suggest that M protein may be present in the virion in close enough proximity to interact with each other and may further have some interactions with glycoprotein and nucleocapsid protein. On the basis of these findings possible roles of M protein in virus assembly were discussed.

Dimethyl Adipimidate↗
PubMed2002

Antigenic heterogeneity amongst the field isolates of Newcastle Disease Virus (NDV) in relation to the vaccine strain. Part II: studies on viruses isolated from domestic birds in Israel.

Forty three Newcastle disease virus (NDV) strains isolated before and during 1997 in Israel from domestic birds were studied by means of the three panels of monoclonal antibodies prepared against all the viral envelope proteins in order to reveal the possible antigenic differences between them and the VH strain used in Israel for poultry vaccination. Three isolates were found to have significant antigenic differences in the hemagglutinin-neuraminidase (HN) and fusion (F) glycoproteins as compared to the vaccine strain. As to the matrix protein, almost all the viruses isolated during the year 1997 were found to have considerable differences from the vaccine strain in two of four antigenic sites.

Animals↗
PubMed1986

Fusion of Newcastle disease virus with liposomes: role of the lipid composition of liposomes.

We demonstrate here that fusion occurs between the membrane of the Newcastle disease virus (NDV) and liposomes. Fluorescence dequenching studies (using Rhodamine-bearing viral envelopes) revealed the mixing of the lipids constituting the viral and liposomal membrane. The digestion of internal viral proteins by trypsin-containing liposomes indicated the mixing of the internal aqueous compartments. This last assay is independent of exchange of lipids between liposomal and viral membrane in the absence of fusion. Investigation of the effects of liposomal composition indicated that the presence of phosphatidylethanolamine and gangliosides are essential to optimize fusion. The fact that the Newcastle disease virus membrane can fuse with liposome also confirms that fusion must be determined by the viral proteins and could be mostly independent of the nature or presence of the host proteins.

Cholesterol↗
PubMed1977

Suppressive effect of Newcastle disease virus on the primary and secondary immune responses of hamsters.

The effect of Newcastle disease virus (NDV) on the formation of 19 S haemolytic plaque-forming cells (HPFC) in the secondary immune response and on the formation of 7 S HPFC in both primary and secondary immune responses of hamsters to sheep red blood cells (SRBC) was studied. The 19 S and the 7 S HPFC in the spleens of hamsters injected with SRBC were demonstrated by the direct and indirect Jerne's method, repectively. A single intraperitoneal injection of NDV 5 days prior to primary immunization markedly depressed the number of 7 S HPFC. NDV administered 5 days prior to the second injection of SRBC resulted in a significant decrease in the number of both 19 S and 7 S HPFC. It is asssumed that interferon induced by NDV is responsible for the suppressive effect of the virus on both the primary and secondary immune responses of hamsters to SRBC.

Animals↗
PubMed1984

Ocular pathogenesis of Newcastle disease virus in rabbits and monkeys.

Studies on the ocular pathogenesis of Newcastle disease virus (NDV) in rabbits and monkeys revealed that conjunctivitis can only be produced by the virus when conjunctival epithelium is traumatized; multiple instillations of large doses of virus over the intact conjunctiva failed to do so. The presence of serum HI antibodies showed no correlation with the clinical form of the disease in rabbits. However, in monkeys, the disease showed a uniform pattern of immune response irrespective of the strain of the virus. Monkeys may be useful for further studies.

Animals↗