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PubMed2004

Importance of serine 200 for functional activities of the hemagglutinin-neuraminidase protein of Newcastle Disease Virus.

Newcastle disease virus (NDV) is an avian paramyxovirus with replication competence in human tumor cells and interesting anti-neoplastic and immune stimulatory properties. In order to increase tumor selectivity of replication, we prepared mutants from the avirulent strain Ulster with monocyclic replication cycle and adapted them for multicyclic replication in human melanoma cells. Two mutants (M1 and M2) showed interesting functional differences: while M2 showed T cell co-stimulatory effects in a tumor-specific cytotoxic T lymphocyte (CTL) assay, M1 did not. A distinct difference of these 2 virus mutants appeared also when testing their capacity to induce interferon-alpha and -beta as well as tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) molecules in human monocytes. Sequence analysis of the hemagglutinin-neuraminidase (HN) molecules of the 2 virus mutants showed 7 non-silent mutational differences. Upon cloning of the HN mutant genes into an expression vector and transfection of cells, only HN derived from M2 (HN-M2) was detected at the cell surface by immunostaining with specific antibodies and showed hemadsorption and neuraminidase activity. In order to define which amino acid was responsible for the loss of functional activity of HN derived from M1 (HN-M1), distinct HN mutants were generated via site-directed mutagenesis and tested. Substitution of serine 200 by a proline abrogated HN expression and its hemadsorption and neuraminidase activities. Molecular modeling revealed that proline 200 in HN influences flexibility of a loop near the entrance to the neuraminidase active site, a function that may be crucial for the functions of this viral protein.

Animals↗
PubMed1993

Retinoic acid enhances killing of neuroblastoma cells by Newcastle disease virus.

Newcastle disease virus (NDV), an avian pathogen, selectively replicates in and kills neuroblastoma (NB) cells, but not normal fibroblasts in vitro and in vivo in nude mice. NDV cytotoxicity towards NB cells is enhanced by N-myc oncogene amplification. To further define the antineoplastic effects of NDV, we examined NDV's interaction with NB cells following short-term exposure to the differentiating agent, all-trans retinoic acid (RA), and to neuraminidase. The human NB cell line IMR-32, after treatment with 50 mumol/L RA, became eight times more sensitive to NDV in a cytotoxicity assay. A time course study to determine the optimal incubation period of IMR-32 cells with RA indicated that a fourfold increase in sensitivity towards NDV killing occurred after only 8 hours of RA incubation prior to addition of virus. Maximal sensitivity was achieved at 24 hours of RA incubation and remained constant for longer incubation periods (up to 72 hours). The sensitization of IMR-32 NB cells to NDV was constant for RA doses between 3 mumol/L and 50 mumol/L. Plaque formation, which indicates replication, virus spread and cytotoxicity by a single infectious virus particle, was also enhanced by RA. This effect does not appear to require N-myc amplification in the target NB cells since RA had similar effects upon the high N-myc (IMR-32) and the low N-myc expressing cells (SK-N-SH). Enhanced sialylation has been shown by others to mediate the growth inhibitory effects of RA on a variety of tumor lines. Removal of sialic acid from the IMR-32 NB cell surface using Clostridium neuraminidase (2.7 mg/mL) inhibited 75% of NDV plaque formation. These results demonstrate that NDV killing of two NB cell lines is enhanced using clinically achievable levels of RA and that sialylation of the NB cell surface is important for virus binding and cytotoxicity.

Cytopathogenic Effect, Viral↗
PubMed1988

Serological response of chickens to oral vaccination with Newcastle disease virus.

Conventional Newcastle disease vaccines are not suitable for application to village chickens in tropical countries of Asia. Trials with food-based vaccines are being initiated and the following experiments were performed to evaluate oral vaccination with Newcastle disease virus. Experimental chickens were vaccinated orally with the avirulent V4 strain of Newcastle disease virus and haemagglutination-inhibition antibody responses were measured. V4 virus was introduced into the crop by tube and total faecal output was collected daily and assayed for Newcastle disease virus. Virus was recovered on Days 5 and 6 after vaccination from most chickens that had received 10(7.4) and 10(6.4) 50% egg-infectious doses (EID50) of virus. There was no recovery of virus from birds receiving a lower dose of vaccine. Groups of chickens kept in cages with wire floors were given various doses of vaccine into the crop. Higher antibody titres were achieved with higher doses of virus. This dose responsiveness was not observed when various doses of vaccine were presented on food pellets and the groups of chickens were kept on concrete floors. Similar antibody responses were then seen with nominal doses of 10(5.2) and 10(8.2) EID50 per bird, possibly as a result of excretion and re-ingestion of the vaccine virus. Spread of the vaccine virus was demonstrated when control chickens and chickens receiving 10(7.7) EID50 of V4 virus on food pellets were housed together on a concrete floor. Similar antibody titres were achieved in both vaccinated and in-contact chickens.

Administration, Oral↗
PubMed2003

Feasibility study to evaluate the correlation between results of a candidate in vitro assay and established in vivo assays for potency determination of Newcastle disease vaccines.

A Newcastle disease virus antigen quantification assay has been developed at CIDC-Lelystad as a candidate in vitro potency test for inactivated Newcastle disease vaccines. In studies performed at CIDC-Lelystad, a high correlation was demonstrated between the results of this candidate in vitro potency assay and the results of the serological potency assay (European Pharmacopoeia monograph 0870; test A). Furthermore, a high correlation between the serological data (Haemagglutination Inhibition-antibody titres) and clinical protection after challenge was demonstrated. The aim of the feasibility study was to confirm the correlation between the results obtained using the candidate in vitro potency assay and the results from both the in vivo potency assays currently prescribed in Ph Eur monograph 0870, in different laboratories and to determine whether a large-scale validation study of the in vitro method should ensue. In the feasibility study three Official Medicines Control Laboratories tested the potency of 5 different inactivated Newcastle disease vaccines and one experimental vaccine, using both of the in vivo methods described in the European Pharmacopoeia and the candidate in vitro method. The 6 vaccine batches represented a quantitative range of Newcastle disease virus antigen content and were produced by different manufacturers. Statistical evaluation of all results indicated that a satisfactory correlation was found in all laboratories between the two types of in vivo tests currently in place, and the candidate in vitro test. An excellent reproducibility of the proposed in vitro method was observed with respect to the ranking of the vaccines included in this study. It is concluded that the results of this feasibility study indicate that a large-scale collaborative study can be organised to validate the in vitro method and the suitability of the reference preparation.

Animal Use Alternatives↗
PubMed1996

Protection of chickens from Newcastle disease by vaccination with a linear plasmid DNA expressing the F protein of Newcastle disease virus.

To evaluate the usefulness of a DNA vaccine for chickens, we constructed a plasmid vector expressing the Newcastle disease virus F protein (NDV-F) under the control of the human cytomegalovirus immediate early enhancer and chicken beta-actin gene promoter. One-week-old chickens injected intramusculary with the circular plasmid DNA did not produce significant levels of antibody against NDV-F. However, two of five birds injected with the linearized plasmid DNA produced high levels of the antibody. Moreover, four of five birds injected with a mixture of the linearized-plasmid and Lipofectin produced the antibody efficiently. At 9 weeks post-injection, chickens were challenged with the velogenic NDV Sato strain. These chickens that had the antibody against NDV-F were protected from lethal NDV challenge. These results demonstrate that the DNA vaccine conferred efficient protection against the disease.

Animals↗
PubMedDate not supplied

Atypical disease produced in chickens by Newcastle disease virus isolated from exotic birds.

Chickens were infected with a Newcastle disease virus (NDV) recovered from exotic birds with severe clinical disease and with lesions characteristic of viscerotropic velogenic Newcastle disease (VVND). The infection in chickens was inconsistently lethal, some infected chickens were not clinically affected, and gastrointestinal involvement was only marginally evident. Pathogenicity of the virus for chickens was not detectably altered by laboratory passage in chickens or by limit dilution passage in chicken embryos. The results suggest that the difference between velogenic NDV pathotypes may not always be distinct and that clinical manifestations of VVND in chickens may not always be predictable based on signs and lesions observed in exotic birds.

Age Factors↗
PubMed2006

Efficacy of live B1 or Ulster 2C Newcastle disease vaccines simultaneously vaccinated with inactivated oil adjuvant vaccine for protection of Newcastle disease virus in broiler chickens.

Two hundred, one-day-old broiler chicks were divided into groups 1, 2 and 3 containing 60, 70 and 70 chicks, respectively. The groups were divided into subgroups of 10 chicks that were vaccinated according to the following scheme: group 1 unvaccinated control, group 2 vaccinated subcutaneously at 1 day old with inactivated oil adjuvant vaccine (IOAV) in combination with live B1 vaccine. Group 3 was vaccinated in the same mode as group 2 with IOAV and live Ulster 2C vaccine. All birds were challenged when they were 28 days old. Mortality rate, body weight gain and feed conversion ratio (FCR) were monitored before and after challenge. All the chickens in group 1 died, indicating that there was no disease resistance of this unvaccinated control group of chickens. Conversely, the monitored disease resistance of chickens in groups 2 and 3 was 68.57% +/- 18.64 and 88.57% +/- 9.00, respectively (P < 0.05). The morbidity of chickens in groups 2 and 3 was 37.89% +/- 14.36 and 14.76% +/- 12.40, respectively (P < 0.05). The body weight gain, feed intake and FCR of group 3 were significantly better than those of group 2 (P < 0.05) during 1-42 days old. The simultaneous vaccination with B1 or Ulster 2C and IOAV of 1-day-old chicks gave some protection of 28-day-old broilers without a booster vaccination.

Animals↗
PubMed1992

A ten-year follow-up on stage II malignant melanoma patients treated postsurgically with Newcastle disease virus oncolysate.

Newcastle disease virus oncolysate was examined as an adjunctive immunotherapeutic agent in the postsurgical management of 83 cases of Stage II malignant melanoma. At this time, all the patients have been under observation for at least 10 years, and over 60% are alive and free of recurrent disease. Older studies in the United States report postsurgical survival figures for Stage II cases of 5-15%. More contemporary studies indicate a 33% survival at 10 years. The unusual disease-free survival periods in the present study, including exceptional survivals in 21 patients with head and neck disease and six cases with cerebral metastases, suggest a unique role for the administration of Newcastle disease virus oncolysate in the management of Stage II malignant melanoma patients.

Adult↗
PubMed1986

Genotypic and phenotypic variation of biotypes coexisting in the Hickman strain of Newcastle disease virus.

Many Newcastle disease virus strains are composed of several biotypes which coexist in the wild and in laboratory cultures. We have studied some of the phenotypic and genotypic properties of 6 virus clones from the coexisting biotypes of the Hickman strain of Newcastle disease virus. These clones were readily distinguishable from the parent virus strain and from each other by their RNA fingerprints. Fingerprints of the most virulent clones (Hi/LC, Hi/MC, and Hi/LR) contained 61% to 77% of the oligoribonucleotides present in the fingerprint of the Hickman parent strain. None of the clones killed chickens as rapidly as did the parent strain, although some clones killed embryonating eggs as rapidly as did the parent strain.

Genotype↗
PubMed1992

Immunization of day-old chickens against Newcastle disease.

The avirulent Newcastle disease virus strain designated NDV-6/10, selected by B. Lomniczi at the Veterinary Medical Research Institute, Hungarian Academy of Sciences, is completely safe for day-old chickens by aerosol vaccination. Aerosol immunization using the Hungarian-made MASTERDROP generator (particle size: maximum 7 microns) caused no vaccination reactions among 206,000 chickens with different maternal antibody levels. Other vaccines given simultaneously did not significantly affect the protection elicited against Newcastle disease (ND). Almost 100% and 90% of the aerosolized chickens survived subcutaneous challenge with 10(6) LD50 NDV at 30 and 50 days old, respectively. A single immunization is sufficient for broilers; however, parent flocks should be revaccinated at 7 so 8 weeks old.

Administration, Inhalation↗
PubMed1995

Brain lesions in chickens experimentally infected with a neuroadapted strain of mesogenic Newcastle disease virus.

Neuroadapted Newcastle disease virus (Q10) was selected by tenth serial passage, in the chicken brain of a mesogenic strain (Q0) originally isolated from quails. Specific pathogen-free birds were inoculated intranasally with one of these viruses. At daily intervals for 7 days and then at 10, 14, and 21 days post-inoculation (PI), two birds from each group were killed and samples of the brain were collected for histopathological and virological examination. Q10 caused severe nonsuppurative encephalitis with nervous signs and high mortality. Lesions characterized by neuronal degeneration and necrosis, perivascular lymphocytic infiltration, and focal or diffuse astrogliosis occurred mainly in the parahippocampal cortex, hippocampus, hyperstriatum, neostriatum, subleptomeningeal and periventricular regions of the cerebrum. Spongy changes with neuronal degeneration and axonal spheroids were also observed in the brain stem of a few cases. The amount of virus in the brain reached a peak on day 4 PI and virus could not be recovered from the brain after 6 days PI. In contrast, Q0 caused nonfatal asymptomatic disease and virus could not be isolated from the brain, sections of which showed only minimal inflammatory changes. This difference in the lesions of the brain might be related to neurovirulence and, neuroadaptation by serial passage may occur by increased efficiency of viral replication in neurons.

Animals↗
PubMed1983

Interferon induction by viruses. X. A model for interferon induction by Newcastle disease virus.

Unirradiated Newcastle disease virus (NDV, strain AV) induced high levels of interferon (IFN) in primary chick embryo cells if the cells were 'aged' in vitro for 6 to 7 days. Dose (multiplicity)-response (IFN yield) curves, carried out in the presence of anti-NDV serum to prevent cycling infection, revealed that stocks of NDV-AV contain about sevenfold more IFN-inducing particles (IFP) than infectious particles (PFP). These non-infectious IFP were responsible for nearly all IFN induction in 'aged' cells, since PFP were determined to be incapable of inducing IFN. In contrast, with mouse L(Y) cells as hosts, about one-third the number of particles as there are PFP appeared to score as IFP. Heat and u.v. radiation (254 nm) inactivated NDV IFP and PFP activity at the same rate whether tested in chick or mouse cells, implying that virion-associated transcription is required to induce IFN. A model is proposed to account for the generation of IFN-inducing particles from infectious NDV following u.v. irradiation, and their subsequent inactivation at high doses of radiation. The model defines a series of u.v. targets in the NDV genome that regulate the expression of IFN-inducing particle activity in 'unaged' chick embryo cells.

Animals↗
PubMed2001

Use of a heteroduplex mobility assay to detect differences in the fusion protein cleavage site coding sequence among Newcastle disease virus isolates.

Newcastle disease virus (NDV) is an economically important pathogen of poultry that may cause clinical disease that ranges from a mild respiratory syndrome to a virulent form with high mortality, depending on an isolate's pathotype. Infections with virulent NDV strains are required to be reported by member nations to the Office of International Epizootes (OIE). The primary determinant for virulence among NDV isolates is the presence or absence of dibasic amino acids in the fusion (F) protein cleavage activation site. Along with biological virulence determinations as the definitive tests, OIE accepts reporting of the F protein cleavage site sequence of NDV isolates as a virulence criterion. Nucleotide sequence data for many NDV isolates recently isolated from infected chickens and other avian species worldwide have been deposited in GenBank. Consequently, viral genomic information surrounding the F protein cleavage site coding sequence was used to develop a heteroduplex mobility assay (HMA) to aid in further identification of molecular markers as predictors of NDV virulence. Using common vaccine strains as a reference, we were able to distinguish virulent viruses among NDV isolates that correlated with phylogenetic analysis of the nucleotide sequence. This technique was also used to examine NDV isolates not previously characterized. We were able to distinguish vaccine-like viruses from other isolates potentially virulent for chickens. This technique will help improve international harmonization of veterinary biologics as set forth by the OIE and the Veterinary International Cooperation on Harmonization of Technical Requirements of Veterinary Medicinal Products. Ultimately, the HMA could be used for initial screening among a large number of isolates and rapid identification of potentially virulent NDV that continue to threaten commercial poultry worldwide.

Amino Acid Sequence↗
PubMed2005

Aflatoxicosis, infectious bursal disease and immune response to Newcastle disease vaccination in rural chickens.

To investigate the immunosuppressive effects of infectious bursal disease virus (IBDV) and aflatoxin in indigenous chickens of Uganda, Newcastle disease (ND) seronegative chicks were randomly allocated to two treatment groups. Group A chicks were injected intramuscularly at the age of 3 weeks every 2 days up to four times with 0.250 mg aflatoxin B1 per bird, group B was infected occulo-nasally with IBDV 3 days prior to vaccination, while group C was left as a control group. All the chicks from the three groups were then vaccinated with Hitchner B1 vaccine at 21 days of age followed by a secondary vaccination with La Sota vaccine 3 weeks later. Humoral and cell-mediated immune responses were assessed by measuring antibody levels and delayed hypersensitivity reaction post vaccination. Growth performance in the three groups was assessed by weekly body weights while evidence of excretion of vaccinal ND virus was detected by reverse transcription-polymerase chain reaction.A significant (P < 0.05) reduction in the haemagglutination inhibition of ND antibody titre following initial priming with Hitchner B1 and subsequent booster with La Sota vaccines and a delayed hypersensitivity test following sensitization with dinitrochlorobenzene showed aflatoxin to be a more potent immunosuppressant than IBDV. Aflatoxin exerted its maximum effects during primary antibody response in the second and third weeks post vaccination. Aflatoxin and IBDV did not affect growth rates (P > 0.05) but prolonged La Sota vaccine virus excretion in faeces. Under our experimental conditions, aflatoxin and IBDV do not significantly affect the immune response of rural chickens to ND vaccination.

Aflatoxin B1↗