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PubMed1984

Vaccination of chickens with newcastle disease and infectious bronchitis vaccines administered singly and in combination.

The reactivity and immune response were determined in chickens following the administration of Newcastle disease (ND) and infectious bronchitis (IB) vaccines given singly and in combination. When given singly, the IB component resulted in an earlier appearance of respiratory signs than when it was combined with ND vaccine. The intensity of the respiratory reaction appeared similar in each case, however. Measured by virus-serum neutralization and from challenge with ND and IB viruses, no evidence of interference in the inducement of immunity was present with either vaccine fraction. The possible effect of vaccine titers and strain characteristics was discussed briefly.

Animals↗
PubMedDate not supplied

[Utilization of poly U-cellulose and poly U-sepharose for the study of virus-specific RNA of Newcastle disease virus].

Comparative characteristics of poly(U)-cellulose and poly(U)-sepharose used for the study of virus-specific 18S RNA of Newcastle disease virus are presented. Upon chromatography of 18S NDV RNA on any of these columns, approximately 60-70% RNA was adsorbed. The sorbing RNA contained in poly(A) 10-12% of the total amount of adenosine, in contrast to RNA not adsorbed on the column in which poly(A) contained 1-2% of the total amount of adenosine in RNA molecule. The analysis in sucrose density gradient and polyacryl amide gel of the RNAs eluated from the columns showed RNA chromatography under the conditions used not to cause its degradation. The advantages and short-comings of the sorbents used in studies of virus-specific RNA are discussed.

Cellulose↗
PubMedDate not supplied

Newcastle disease virus-induced apoptosis in PC12 pheochromocytoma cells.

The avian paramyxovirus Newcastle disease virus (NDV) causes severe infections in birds. It is essentially nonpathogenic in rodents and human beings but was found to have an oncolytic potential against certain types of human malignancies. An attenuated NDV vaccine (designated MTH-68/H) was found to cause regression of various human tumors, but the mechanism of its oncolytic action and its selectivity toward malignant cells remain poorly understood. NDV was reported to cause apoptotic death in several avian cultured cell types. Programmed cell death may thus be the basis for the oncolytic effect of NDV vaccines. To test this possibility, we chose the PC12 rat pheochromocytoma cell line, a widely used model system for apoptosis. The MTH-68/H vaccine was found to cause apoptotic death of PC12 cells in a dose-dependent manner. A brief exposure of cells to the virus was found to trigger the apoptotic response. Cell death induced by the vaccine was not accompanied by significant alterations in the major mitogen-activated protein kinase pathways of these cells. Apoptotic DNA fragmentation was not affected by stimulating growth factor pathways or signaling mechanisms mediated by protein kinase C or the second messenger, calcium. In contrast, stimulation of protein kinase A by cyclic adenosine monophosphate analogs gave partial protection against the virus. PC12 cells thus provide a useful model system to study the effects of NDV on cell survival at the molecular level.

Animals↗
PubMed1987

The nucleotide sequence of the gene encoding the Newcastle disease virus membrane protein and comparisons of membrane protein sequences.

The nucleotide sequence of a cloned cDNA copy of the mRNA encoding the Newcastle disease virus (NDV) membrane (M) protein was determined. A single open reading frame in the sequence encodes a protein of 364 amino acids with a calculated mol wt of 39,742. The predicted protein sequence does not contain extensive hydrophobic regions. The sequence does contain eight pairs of basic amino acid residues, five of which are located in the carboxyl-terminal half of the sequence. Comparisons of the NDV M protein sequence with other paramyxovirus M protein sequences reveals little homology common to all sequences. There is only 17% homology with Sendai virus M protein. A short region of homology with the VSV M protein sequence, was, however, found.

Amino Acid Sequence↗
PubMed1985

Studies on the development of pelleted Newcastle disease virus (NDV) vaccine.

White leghorn chickens (7-10 weeks old) were orally infused with varied doses of lentogenic and mesogenic strains of Newcastle disease virus. The antibody response of the chickens was monitored by haemagglutination inhibition tests weekly for 4 weeks postinfusion. A high dose of virus (10(9) EID50) was necessary to induce a serological response typical of chickens that resist challenge. Sera from chickens infused with low doses (10(5) EID50) of the virus only showed marginal haemagglutination inhibiting activity. Trypsin levels as high as 0.35 mg ml-1 were detected from duodenal secretions and are assumed to be responsible in part for viral inactivation. Fasting (no food and water) the chickens beyond 12 h did not appear to increase antibody response to low titre virus.

Animals↗
PubMed1987

Antigenic characterization of the internal proteins of Newcastle disease virus by monoclonal antibodies.

We have prepared and characterized monoclonal antibodies against the three internal structural proteins, M, P and NP, of Newcastle disease virus. At least two non-overlapping antigenic sites were delineated on the M protein, four on the P, and two on the NP by competitive binding assay. One of the two non-overlapping antigenic sites on the M protein was found to be a cluster of at least three distinct epitopes. Enhancement of antibody binding by the binding of a second antibody was observed with the M protein. The reactivity of these monoclonal antibodies with heterologous strains was studied by enzyme-linked immunosorbent assay. The results indicated that there are both highly conserved antigenic sites and those subject to remarkable change on both M and P proteins. On the other hand, NP appeared to be antigenically more stable.

Antibodies, Monoclonal↗
PubMed1976

Immunoglobulins of the chicken antibody to Newcastle disease virus (Mukteswar and F strain).

The qualitative and quantitative development of chicken immunoglobulins in response to R2B (Mukteswar) and F strain of Newcastle disease virus was studied. One primary (R2B) and two secondary (R2B - R2B, F - R2B) vaccination trials were conducted. Sera was collected at weekly intervals and analyzed. There was an increase in total serum protein content in parallel to an increase in serum neutralizing (SN), hemagglutination inhibition (HI), and precipitating antibodies. The SN, HI and precipitating activities were detected both in IgM and IgG immunoglobulins when sera were treated with mercapto-ethanol. However, Sephadex G-200 fractionated sera showed only SN activity in the IgM fraction, whereas the IgG fraction showed both HI and SN activity. Serum IgM antibodies appeared during the first week following vaccination, then diminished, then rose again following secondary vaccination. Apparently, immunoglobulin induction and serological activities were not significantly influenced by the age of the chickens.

Animals↗
PubMed1996

Detection of extraneous agents in vaccines using the polymerase chain reaction of Newcastle disease virus in poultry biologicals.

Reverse transcription-polymerase chain reaction (RT-PCR) in poultry vaccine was applied to the detection of Newcastle disease virus (NDV), using two primer pairs spanning the cleavage site of the FO fusion protein coding sequence. Amplification of a specific cDNA segment was possible from live and inactivated, oil-adjuvanted NDV vaccines without previous treatment. The RT-PCR was able to detect between 5 x 10(2) EID50 (in live vaccine preparations) and 10(5) EID(50) or 0.056 haemagglutinating units of NDV (in inactivated vaccine preparations). In addition, live vaccine preparations were inactivated with beta-propiolactone (beta-PL). Amplified cDNA was obtained after treatment with 0.1% beta-PL, whereas at a concentration of 1% or 10% no specific bands were visible in the agarose gel. These results demonstrate the applicability of the method for the control of poultry vaccines by ensuring the absence of extraneous agents.

Animal Testing Alternatives↗
PubMed1991

Effect of vitamin A deficiency on the activity of macrophages in Newcastle disease virus-infected chickens.

The effect of vitamin A deficiency on the activity of peritoneal macrophages (PM) was investigated in noninfected and Newcastle disease virus (NDV)-infected chickens. Day-old chickens with limited vitamin A reserves were fed diets containing either marginal (120 retinol equivalents (RE)/kg) or adequate (1200 RE/kg) levels of vitamin A. At 4 weeks of age, half of the chickens in each group were infected with the La Sota strain of NDV and PM were isolated 11 or 12 days later. These were used for counting the uptake of fluorescein isothiocyanate-labeled yeast cells as an indicator of phagocytic activity and for measuring the reduction of nitroblue tetrazolium (NBT), which provides an estimate of oxygen-dependent killing of microorganisms. Vitamin A deficiency impaired NBT reduction and, to a lesser extent, phagocytosis in both infected and noninfected chickens. NDV infection increased phagocytosis and NBT reduction in normal and, to a lesser extent, in vitamin A-deficient chickens.

Animals↗
PubMed1997

Assembly of recombinant Newcastle disease virus nucleocapsid protein into nucleocapsid-like structures is inhibited by the phosphoprotein.

A recombinant baculovirus expressing the nucleocapsid gene (NP) of Newcastle disease virus (NDV), a member of the genus Rubulavirus, has been generated and shown to express the native protein to high levels in insect cells. In contrast to the NP protein of the rubulavirus human parainfluenza virus 2, the NDV protein has been demonstrated by electron microscopy and caesium chloride gradient analysis to be capable of self-assembly in vivo to form nucleocapsid-like structures in the absence of other NDV proteins. These structures, which contained RNA that was resistant to micrococcal nuclease digestion, were also observed when the protein was expressed in E. coli, a phenomenon which was not inhibited by the presence of a 40 amino acid fusion region at the amino terminus of the protein. Further, the formation of these structures was inhibited by the co-expression of the phosphoprotein (P). Therefore, we conclude that the P protein acts as a chaperone, preventing uncontrolled encapsidation of non-viral RNA by NP protein.

Animals↗
PubMed1980

Relationship between enhanced macrophage phagocytic activity and the induction of interferon by Newcastle disease virus in mice.

The relationship between phagocytic activity of peritoneal macrophages and serum interferon (IF) titers was evaluated in mice challenged with Newcastle disease virus (NDV). Time course studies indicated peak serum IF titers between 6 and 12 hr, whereas Fc receptor-mediated macrophage phagocytosis was maximal 18 hr after viral administration. Both responses decreased in parallel as the inoculated dose of the virus was reduced. Splenectomy, shown by others to decrease the NDV-induced serum IF titers, significantly decreased the stimulation of phagocytosis. The role of T cells in the response to the virus was studied with nude mice raised under germfree conditions. NDV-induced serum IF titers and macrophage phagocytosis were both diminished in BALB/c nudes compared with their heterozygous littermates. Both responses could be partially restored by transfer of thymocytes obtained from heterozygous mice. The results provide further evidence that in vivo macrophage stimulation by NDV is mediated by induced IF. The experiments with nude mice also indicate that the IF response to NDV is regulated by T lymphocytes.

Animals↗
PubMed2005

Monoclonal antibody routinely used to identify avirulent strains of Newcastle disease virus binds to an epitope at the carboxy terminus of the hemagglutinin-neuraminidase protein and recognizes individual mesogenic and velogenic strains.

Newcastle disease virus (NDV) strains are classified as having high (velogenic), intermediate (mesogenic), or low (lentogenic) pathogenesis and virulence in chickens. Recent studies have established that the hemagglutinin-neuraminidase (HN) protein plays an important role in viral tropism and virulence. A monoclonal antibody (AVS-I) has previously been shown to be specific for lentogenic strains of NDV (Srinivasappa et al., Avian Dis. 30:562-567, 1986) and is routinely used to identify these strains. We have used competition antibody binding assays with a previously characterized panel of monoclonal antibodies, binding to chimeric HN proteins, and the characterization of an escape mutant to localize the binding site of AVS-I to the extreme carboxy terminus of the protein. In addition, we have shown that AVS-I does recognize at least one mesogenic strain and one velogenic strain of the virus, calling into question the potential of this antibody as a diagnostic reagent for avirulent NDV strains.

Antibodies, Monoclonal↗