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PubMed2001

Membrane glycoproteins of Newcastle disease virus: nucleotide sequence of the hemagglutinin-neuraminidase cloned gene and structure/function relationship of predicted amino acid sequence.

The nucleotide sequence of the glycoprotein hemagglutinin-neuraminidase (HN) gene of the Newcastle disease virus (NDV) strain Clone-30 has been determined. The open reading frame of the HN gene contains 1731 nucleotides and encodes a protein of 577 amino acids. Three highly conserved patterns among all paramyxovirus HN glycoproteins, and one additional conserved species-specific region are present. The protein contains five potential N-glycosylation sites, all but one located in the C-terminal external domain. The secondary structure prediction shows that the C-terminal external domain is mostly arranged in beta-sheets, while alpha-helices are predominantly located in the N-terminal domain. The nucleotide sequence data of the HN gene reported in this paper has been deposited in the GenBank database, under accession number AF098289.

Amino Acid Sequence↗
PubMed1995

Receptors for the V4 strain of Newcastle disease virus in the digestive tract of chickens.

Enterocytes were detached from various parts of the digestive tract of chickens by treatment with DTT or with hyaluronidase. Isolated enterocytes were exposed to suspensions of the V4 strain of Newcastle disease virus (NDV). Removal of virus from the supernatant fluid was taken as evidence of binding of virus to enterocytes and residual virus was measured both by infectivity assay and by ELISA. Enterocytes from duodenum, jejunum, ileum, caecum, and rectum bound the virus; enterocytes from oesophagus, crop and proventriculus did not.

Animals↗
PubMed1999

Comparative characteristics of the Israeli Newcastle disease virus field strains by means of a wide panel of monoclonal antibodies against hemagglutinin-neuraminidase glycoprotein.

Fourteen mouse monoclonal antibodies (MAB) were tested for their ability to react with 15 reference and 52 local Newcastle disease virus (NDV) strains isolated in Israel during the last decade from feral birds. All the field isolates had no antigenic difference when examined by classic serological tests. However, MAB-mediated analysis revealed wide antigenic heterogeneity amongst the studied viruses. By the pattern of the MAB reactivity, all the isolates could be distributed into 13 groups.

Animals↗
PubMed2004

High cell surface expression of Newcastle disease virus proteins via replicon vectors demonstrates syncytia forming activity of F and fusion promotion activity of HN molecules.

For functional studies, the hemagglutinin-neuraminidase (HN) and the fusion protein (F) of Newcastle disease virus (NDV) were expressed in BHK cells using two vectors which are based on the Semliki Forest virus (SFV) replicon. The first system of high protein expression works by transfection of RNA which before has been in vitro transcribed from a vector containing the gene for the SFV self-amplifying replicase (REP) and a foreign gene using the SP6 promoter. A high level of protein (HN or F) expression was detected 18-20 h after transfection. To study the host range of this expression system, a panel of different cell lines were compared for transfections with SFV RNA. A wide range of expression efficiency was observed, the highest being BHK cells. The second system is based on a DNA plasmid in which the SFV-REP and a foreign gene are expressed in cells under the transcriptional and translational control of the cytomegalovirus immediate-early enhancer T7 promoter. DNA-electroporated BHK cells expressed also high levels of the recombinant proteins but at a delayed time point (40-48 h) as compared with the corresponding RNA. Co-expression of the two NDV proteins, HN and F, via this DNA vector in the same cells led to syncytium formation in the cell monolayer, showing that both proteins expressed in this way, were functionally active. F alone, expressed via this vector, displayed residual fusion activity suggesting its proteolytic cleavage and its functional independence from HN.

Animals↗
PubMed1994

Induction of cytokines and cytotoxicity against tumor cells by Newcastle disease virus.

The use of NDV as biological adjuvant in vaccines against human cancer is still actual in several clinical treatment protocols. In this study, we have investigated in vitro-effects of Newcastle disease virus (NDV) strain 73-T on isolated mononuclear blood cells and cultured tumor cells. Cellular cytotoxicity of PBMC freshly isolated from healthy donors against tumor cells was enhanced significantly (p < 0.01) after coincubation of NDV with effector cells. NDV failed to enhance cytotoxicity of effector cells when PBMC were stimulated three days with 500 IU recombinant interleukin-2 (rIL-2) per ml prior to coincubation with the virus. No significant enhancement of cellular lysis was seen when only target cells were coincubated with NDV. As shown by depletion of various lymphocyte subsets, NK cells were the predominant mediator of lysis. Enhancement of cytotoxicity correlated with the induction of interferon-alpha (IFN-alpha) in PBMC by NDV. NDV also induced high amounts of tumor necrosis factor-alpha (TNF-alpha) in PBMC. Induction of interferon-gamma (IFN-gamma) was weak. A direct cytopathic effect (CPE) of NDV on different target cells was detected by colorimetric measurement of metabolic cell activity. The human tumor cell lines A-498, A-704, Caki-1, Caki-2, and K-562 and the fibroblast line MRC-5 showed progressive cellular destruction 48 h after infection with NDV, whereas PBMC and Daudi cells remained unaffected during the observation period. The nontransformed monkey kidney cell line CV-1 and the transformed monkey kidney cell line COS-1 were both lysed by NDV with marginal difference in time course of CPE. Our results indicate a reasonable potential of pleiotropic modifications of the immune response against tumors by NDV.

Adjuvants, Immunologic↗
PubMed1994

Complete regression of human neuroblastoma xenografts in athymic mice after local Newcastle disease virus therapy.

BACKGROUND: Neuroblastoma is the most common pediatric extra-cranial solid cancer. Using conventional therapies, children older than 1 year of age with advanced neuroblastoma have a poor prognosis. The development of new approaches for treating such children with neuroblastoma continues to be one of the most important goals today in pediatric oncology. Despite numerous anecdotal reports of human tumor regression during viral infections, the use of viruses to directly lyse neuroblastoma cells has never been reported as a potential therapy. Newcastle disease virus (NDV) has been shown to replicate in and kill cultured human and rat neuroblastoma cells but not normal human fibroblasts. PURPOSE: Our purpose was to determine if this selective killing of human neuroblastoma (IMR-32) cells is maintained during the in vivo treatment of established tumors. METHODS: Two experiments were performed using NDV strain 73-T. Athymic mice with subcutaneous IMR-32 human neuroblastoma xenografts (6-12 mm) were treated intralesionally with live NDV, UV-inactivated NDV, or phosphate-buffered saline (PBS). To study virus replication in situ, mice were given intratumoral or intramuscular injections of NDV. These mice were then killed at various times, and the amount of infectious virus present in tumor or muscle was determined. RESULTS: After one injection of live NDV, 17 of 18 tumors regressed completely, whereas rapid tumor growth occurred in all 18 mice treated with PBS and in all nine mice treated with UV-inactivated NDV (P < .0001). The one tumor that showed only a partial response to a single injection regressed completely after a second NDV treatment. Six months following virus-induced regression, only one tumor had recurred. No significant acute or chronic side effects of live NDV were noted in athymic mice given doses up to 500 times that used in this study. Virus levels increased more than 80-fold between 5 and 24 hours in virus-injected tumors (P < .04), while no infectious virus was produced in NDV-injected muscle tissue. CONCLUSIONS: NDV 73-T appears to replicate selectively in human IMR-32 neuroblastoma xenografts, leading directly to a potent antitumor effect as demonstrated by long-lasting, complete tumor regression occurring after a single local injection of virus. IMPLICATION: These experiments may provide an important step in the development of new therapeutic approaches to challenging cancers such as neuroblastoma.

Animals↗
PubMed2003

Newcastle disease virus and Chlamydia psittaci in free-living raptors from eastern Germany.

Organ samples from free-living raptors from the federal states of Berlin and Brandenburg in eastern Germany were tested for Newcastle disease virus (NDV; n = 331) and Chlamydia psittaci (n = 39) by polymerase chain reaction (PCR). In 18 individuals NDV nucleic acids were detected. These samples originated from barn owls (Tyto alba; n = 15, 28%), tawny owl (Strix aluco; n = 1, 5%), common buzzard (Buteo buteo, n = 1, 1%), and European kestrel (Falco tinnunculus; n = 1, 4%). In 29 (74%) of 39 samples C. psittaci was detected. Chlamydia psittaci is common in free-living birds of prey in the investigated area.

Animals↗
PubMed1991

The serological responses of chickens to mass vaccination with a live V4 Newcastle disease virus vaccine in the field and in the laboratory. 1. Meat chickens.

Meat chickens on commercial broiler farms were vaccinated once at 1 to 15 days of age with a live V4 Newcastle disease virus (NDV) vaccine administered by drinking water, aerosol or coarse spray. Hatchmates were housed and similarly vaccinated in laboratory isolation pens. Samples of birds were bled at weekly to fortnightly intervals and the serums tested for haemagglutination inhibiting antibody to NDV. Log2 mean titres of up to 6.26, and assumed protection levels (based on the percentage of birds with log2 titres of 4 or greater) of up to 89%, were obtained in field trials within 4 weeks of vaccination. Differences were observed between the results obtained from parallel field and laboratory trials. The presence of maternal NDV antibody reduced the response to vaccination. The results show that this V4 vaccine can produce an adequate serological response following mass administration to Australian meat chickens housed under commercial conditions.

Aerosols↗
PubMed1994

Protective effect of individual glycoproteins of Newcastle disease virus expressed in insect cells: the fusion protein derived from an avirulent strain had lower protective efficacy.

Recombinant hemagglutinin-neuraminidase (rHN) and fusion (rF) glycoproteins of virulent and avirulent strains of Newcastle disease virus (NDV) expressed by using baculovirus expression system were used to investigate their protective immunization effects in chickens. The efficacy of immunization with these recombinant proteins was evaluated by challenge infection. The chickens immunized with either rHN or rF protein of a virulent strain or rHN protein of an avirulent strain were completely protected from the lethal infection of virulent NDV. On the other hand, the rF protein of an avirulent strain, in which precursor F protein was not cleaved, showed lower protective effects. Significant levels of specific antibodies against respective proteins were detected in sera from survivors, whereas relatively lower levels of antibodies were found in chickens which were killed by challenge infection. These data indicate that either HN or F protein alone could induce protective immune responses and the cleavage of F protein might be important for its immunological potential.

Animals↗
PubMed1985

Antibody response in partridge (Perdix perdix L.) 1. Effect of sex and age on the immune response to sheep red blood cells (SRBC), Newcastle disease virus (NDV) and Brucella abortus (Buck 19).

To ascertain the immune response capacity of Partridges to classical experimental and natural antigens, dose and time response curves to sheep red blood cells (SRBC), Newcastle disease virus (NDV) and Brucella abortus (Buck 19 strain), were determined in two strains reared in two different farms. 1 ml of 20% SRBC, 10(9) NDV particles and 10(7) Buck 19 microorganisms, gave optimal antibody responses seven (SRBC and Buck 19) or fourteen days after immunization. As far as SRBC is concerned, the antibody response was sex independent, but declined markedly in partridges older than 1 year. The immunization schedule used by us allow the identification of high and low responder animals. In fact, highly significant correlation in individual partridges between antibody titers obtained after two successive immunizations to an optimal SRBC dose given three weeks apart, was found. This fact, together with the observed great variability in the antibody response to the three antigens considered, should allow a selection for high and low responder lines, in order to improve the fitness toward environmental pathogens.

Age Factors↗
PubMed2000

Detection and quantification of antibodies to Newcastle disease virus in ostrich and rhea sera using a liquid phase blocking enzyme-linked immunosorbent assay.

A liquid phase blocking ELISA (LPB-ELISA) was adapted for the detection and quantification of antibodies to Newcastle disease virus. Sera from vaccinated and unvaccinated commercial flocks of ostriches (Struthio camelus) and rheas (Rhea americana) were tested. The purified and nonpurified virus used as the antigen and the capture and detector antibodies were prepared and standardized for this purpose. The hemagglutination-inhibition (HI) test was regarded as the reference method. The cutoff point for the LPB-ELISA was determined by a two-graph receiver operating characteristic analysis. The LPB-ELISA titers regressed significantly (P < 0.0001) on the HI titers with a high correlation coefficient (r = 0.875). The two tests showed good agreement (kappa = 0.82; P < 0.0001), relative sensitivity (90.91%) and specificity (91.18%), and accuracy (91.02%), suggesting that they are interchangeable.

Animals↗
PubMed2000

A single amino acid change in the Newcastle disease virus fusion protein alters the requirement for HN protein in fusion.

The role of a leucine heptad repeat motif between amino acids 268 and 289 in the structure and function of the Newcastle disease virus (NDV) F protein was explored by introducing single point mutations into the F gene cDNA. The mutations affected either folding of the protein or the fusion activity of the protein. Two mutations, L275A and L282A, likely interfered with folding of the molecule since these proteins were not proteolytically cleaved, were minimally expressed at the cell surface, and formed aggregates. L268A mutant protein was cleaved and expressed at the cell surface although the protein migrated slightly slower than wild type on polyacrylamide gels, suggesting an alteration in conformation or processing. L268A protein was fusion inactive in the presence or absence of HN protein expression. Mutant L289A protein was expressed at the cell surface and proteolytically cleaved at better than wild-type levels. Most importantly, this protein mediated syncytium formation in the absence of HN protein expression although HN protein enhanced fusion activity. These results show that a single amino acid change in the F(1) portion of the NDV F protein can alter the stringent requirement for HN protein expression in syncytium formation.

Alanine↗
PubMed2002

Human tumor cell infection by Newcastle Disease Virus leads to upregulation of HLA and cell adhesion molecules and to induction of interferons, chemokines and finally apoptosis.

In order to analyse immune-stimulatory effects of infection of human tumor cells with Newcastle Disease Virus (NDV), gamma-irradiated human breast carcinoma, colon-carcinoma or glioblastoma cells from defined cell lines were modified either by true infection with live virus or by cell surface adsorption of UV-inactivated replication deficient virus. Modification with live but not inactive NDV induced in all human tumor cells IFN-beta and the chemokines RANTES and IFN-gamma-inducible protein-10 (IP-10). In addition, infection by live NDV induced upregulation of HLA-ABC-molecules in all tumor lines tested and HLA-DR molecules in breast carcinoma lines. Two cell adhesion molecules, ICAM-I (CD54) and LFA-3 (CD58), were also upregulated on human tumor cells after infection with live NDV. When infection of MCF-7 breast carcinoma cells by NDV was performed in the presence of neutralizing anti-IFN-beta antibodies no upregulation of HLA molecules was observed suggesting an important role of IFN-beta in this process. Forty-eight to 72 hours after infection of the irradiated tumor cells with live NDV, many tumor cells were dead or in early or late stages of apoptosis. These results provide explanations for the function of the virus-modified autologous tumor vaccine ATV-NDV with which promising clinical results have already been obtained.

Apoptosis↗
PubMedDate not supplied

Increased mortality in turkeys selected for increased body weight following vaccination with a live Newcastle disease virus vaccine.

Candidate male and female breeders from nine genetic lines of turkeys that were reared intermingled, with the sexes housed in different buildings on the same farm, were vaccinated with a live Newcastle disease virus vaccine (B1 type, LaSota) just prior to the commencement of egg production. In 1999, an average mortality for all lines of 5.8% occurred during the 10 days immediately following vaccination and the level of mortality varied among lines. Mortality was, in general, greater in large-bodied lines than in small-bodied lines. Affected birds exhibited gross multiple areas of focal necrosis in the liver and spleen and congestion of the heart and lungs. The percentage mortality occurring following similar vaccination in 2000 averaged 2.6 for the 10 days following vaccination and mortality was greater (P < or = 0.05) in one line (F line) than the other genetic groups and higher in females than in males. Mortality in the F line, selected for increased body weight and known to be susceptible to various diseases, averaged 15.1% for both years. Attempts failed in both years to isolate Pasteurella multocida or other bacteria. There was a positive correlation between increased body weight and increased mortality following vaccination with the live LaSota vaccine.

Animals↗
PubMed1987

Selection for early responsiveness of chicks to Escherichia coli and Newcastle disease virus.

A broiler chicken population was divergently selected for high or low early immune responses to Escherichia coli and to Newcastle disease virus (NDV) vaccines. Four selection cycles were performed in one replicate, and a single cycle in a second replicate. Selection was based on sire-family averages of a titer index (TI) calculated as the mean titer of antibodies produced by offspring vaccinated with either E. coli or NDV at 18 or 10 days of age, respectively. After the first selection cycle, TI of the early-high (EH) line were 22 and 38% greater than those of the early-low (EL) line in Replicates A and B, respectively. After four selection cycles, the average immune response to E. coli and NDV of Line EH exceeded that of Line EL by 68%. Viability was greater in the EH than in the EL line. Realized family heritabilities were .72 and .67 in Replicates A and B, respectively and the levels of response to the two antigens were not genetically correlated. The immune response of the EH line developed earlier than that in the EL line as shown by fewer nonresponders against E. coli and the higher response of this line against the two antigens at young ages. Mortality after challenge at 18 days of age and general mortality from hatching to 20 wk of age was lower in Line EH than in Line EL. Body weights at 7 wk were higher in EH than EL chicks.

Animals↗
PubMed1982

[Demonstration of Newcastle disease virus by enzyme-linked immunosorbent assay].

Investigations were carried out with regard to the application of the sensitive, rapid, and effective enzyme-linked immunosorbent assay (ELISA) in the demonstration of the Newcastle disease virus (NDV). Used was the peroxydase-marked immunoglobulin fraction of the anti-DV rabbit serum in the microconditions of the double-antibody method. The demonstration of the NDV was in a cultivation liquid of NDV-infected BHK21 cells. As control served intact cells as well as strains Roussev and La Sota of NDV pruified through ultracentrifugation on an isokinetic saccharose gradiend. It is stated that ELISA is much more sensitive than the hemagglutination test and the agar gel immunodiffusion test.

Animals↗
PubMedDate not supplied

Effect of negative air ionization on airborne transmission of Newcastle disease virus.

Four-week-old mixed-sex White Rock chickens were used in four experiments to determine the effect of negative air ion enrichment on airborne transmission of the Roakin strain of Newcastle disease virus (NDV). The experiments were conducted in specially constructed airborne disease transmission cabinets in which donor (upwind) chickens cannot contact susceptible (downwind) chickens because of physical separation by a "no man's land." Temperature and humidity were computer-controlled at 26.7 C and 50% relative humidity, and ventilation rates were manually adjusted from 0.34 to 1.36 m3/min (12 to 48 ft3/min). Donor chickens were inoculated with Roakin NDV by eyedrop and intranasal routes and placed in the upwind end of each cabinet. One day later, susceptible chickens were placed in the downwind end. Seroconversion (> or = 1:10 NDV hemagglutination-inhibition titer) was considered evidence of infection from inoculation (upwind) or airborne transmission (downwind). Commercial air ion generators were used either in the ends or in the "no man's land" of the treatment cabinets and operated at power supply voltages ranging from -8kV direct current to -15 kV direct current. The use of negative air ion generators reduced airborne transmission an average of 6.6% to 27.7% compared with the control cabinets. Significant (P < or = 0.05) reductions in transmission were obtained with some treatments. The greatest reduction in transmission was obtained with the higher power supply voltages (13.8% reduction) and when the ionizers were placed in the "no man's land" (27.7% reduction) between the upwind and downwind chickens.

Air Ionization↗