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PubMed1992

Newcastle disease virus selectively kills human tumor cells.

Newcastle disease virus (NDV), strain 73-T, has previously been shown to be cytolytic to mouse tumor cells. In this study, we have evaluated the ability of NDV to replicate in and kill human tumor cells in culture and in athymic mice. Plaque assays were used to determine the cytolytic activity of NDV on six human tumor cell lines, fibrosarcoma (HT1080), osteosarcoma (KHOS), cervical carcinoma (KB8-5-11), bladder carcinoma (HCV29T), neuroblastoma (IMR32), and Wilm's tumor (G104), and on nine different normal human fibroblast lines. NDV formed plaques on all tumor cells tested as well as on chick embryo cells (CEC), the native host for NDV. Plaques did not form on any of the normal fibroblast lines. To detect NDV replication, virus yield assays were performed which measured virus particles in infected cell culture supernatants. Virus yield increased 10,000-fold within 24 hr in tumor and CEC supernatants. Titers remained near zero in normal fibroblast supernatants. In vivo tumoricidal activity was evaluated in athymic nude Balb-c mice by subcutaneous injection of 9 x 10(6) tumor cells followed by intralesional injection of either live or heat-killed NDV (1.0 x 10(6) plaque forming units [PFU]), or medium. After live NDV treatment, tumor regression occurred in 10 out of 11 mice bearing KB8-5-11 tumors, 8 out of 8 with HT-1080 tumors, and 6 out of 7 with IMR-32 tumors. After treatment with heat-killed NDV no regression occurred (P less than 0.01, Fisher's exact test). Nontumor-bearing mice injected with 1.0 x 10(8) PFU of NDV remained healthy. These results indicate that NDV efficiently and selectively replicates in and kills tumor cells, but not normal cells, and that intralesional NDV causes complete tumor regression in athymic mice with a high therapeutic index.

Animals↗
PubMed1998

Comparison of various diagnostic methods in characterizing Newcastle disease virus isolates from Desi chickens.

Eleven Newcastle disease viruses (NDV), isolated from apparently healthy and ailing Desi chickens were subjected to both conventional and modern characterization techniques. The virulence and strain differentiating experiments placed 10 isolates in the velogenic group and one in the mesogenic group. In MDBK cells, 9 isolates produced characteristic cytopathogenic effects up to 5 and 2 up to 3 passages. Molecular characterization with a 21-mer oligonucleotide probe placed all the isolates in the velogenic/mesogenic group. The results of this study clearly indicated that the isolates obtained are either velogenic or mesogenic but not lentogenic.

Animals↗
PubMed2006

A cross-sectional survey of Australian chicken farms to identify risk factors associated with seropositivity to Newcastle-disease virus.

Several outbreaks of virulent Newcastle-disease occurred in Australia in 1998-2000. We conducted a cross-sectional survey of 753 Australian chicken farms to identify risk factors associated with the seroprevalence of chicken flocks with Newcastle-disease virus (NDV). We had a 99.7% response rate to the survey and the overall prevalence of NDV seropositive farms was 39.8%. Associations were analysed for the layer, chicken-meat and breeder production sectors in sector-specific logistic-regression models using 187, 198 and 146 farms, respectively. In the layer sector, increased risk of seroprevalence was associated with increasing age of the chickens, and decreased risk when the nearest-neighbour poultry farm was >10 km distant (odds ratio (OR)=0.30). In the chicken-meat sector, increased risk of seroprevalence was associated with location in the Sydney basin (OR=13.67), eastern Victoria (OR=26.10) or western Victoria (OR=5.43), and decreased risk when the nearest-neighbour poultry farm was greater than 0.5 km distant (OR=0.34). In the breeder sector, increased risk of seroprevalence was associated with increasing age of the chickens, the presence of wild birds on the farm (OR=5.28) and location in eastern Victoria (OR=16.19). A conditional logistic-regression for 112 pairs of farms matched for age, survey region and production sector identified a distance of >1.0 km to the nearest-neighbour poultry farm (OR=0.24) and ownership by owner 2 (OR=0.02), owner 5 (OR=0.11) or owner 9 (OR=0.25) as significant in reducing the risk of NDV seroprevalence. Our survey found that high levels of biosecurity and hygiene practices had been adopted by most farms.

Age Factors↗
PubMed1986

Identification of the sequence content of four polycistronic transcripts synthesized in Newcastle disease virus infected cells.

During infection, the Newcastle disease virus (NDV) genome is transcribed to produce 5 to 7 species of polycistronic messenger RNA (Wilde and Morrison, J. Virol. 51, 71-76) in addition to the well characterized monocistronic messenger RNA. To identify the specific sequences present in each of the polycistronic RNA species, cDNA clones generated by reverse transcription of NDV mRNAs were characterized and used as probes on Northern blots of total NDV cytoplasmic RNA. By this method, it was shown that four of these large RNA species are polycistronic transcripts containing sequences from two genes: one species contains nucleocapsid protein (NP) and phosphoprotein (P) gene sequences; another, P and membrane protein (M) gene sequences; another, M and fusion protein (F0) gene sequences; and another, F0 and hemagglutinin-neuraminidase protein (HN) gene sequences. The existence of these transcripts yields a transcription map order of NP, P, M, F0, HN. The remaining RNA bands may be composed of at least three different polycistronic transcripts, each of which represents transcription through three adjacent genes.

Animals↗
PubMed1981

[Comparison of the thermostability of Newcastle disease virus strains isolated in Slovakia].

Reference strains of Newcastle disease virus of different virulence (two lentogenic, two mesogenic and three velogenic), isolated in Czechoslovakia and other countries, were compared with 11 field strains isolated in Slovakia in 1973, 1977, 1979 and 1980 as to the stability of their infectivity for cell cultures and as to their hemagglutination activity at a temperature of 56 degrees C for 120 minutes. The inactivation curves indicate that ten strains belong to the group of velogenic viruses and one of them is lentogenic. Although the data on hemagglutinin thermostability and infectivity do not suffice to characterize the virus strain, it is possible, by comparing the inactivation curves determined by the described method, to differentiate the field strains of Newcastle disease virus as lentogenic, mesogenic and velogenic. Some instability of the relationship between the thermostability and virulence of the virus is ascribed to the heterogeneity of the virus population.

Animals↗
PubMed1979

[Serological response and the viral carrier state in chickens infected with the pathogenic Newcastle disease virus].

Three groups of susceptible chickens were treated with Newcastle Disease vaccine, as follows: group I--orally, strain La Sota; group II--aerosole treatment, strain La Sota and group III--intramusculary, Komarov's vaccine. Varying HI antibody levels were observed following vaccine application. The average antibody titer of group I and II experimental chickens (40 in number for each group) was 25 days post vaccination respectively log2 = 3.47 and log2 = 6.2 while of group III (50 chickens) it was 14 days post vaccination log2 = 8.4. Aerosole challenge with velogenic viscerothropic strain of Newcastle Disease virus caused a sharp change in serum antibody titer. The low antibody titer in group I rose quickly post challenge while in groups II and III, on the contrary, it fell sharply. This characteristic dynamics is proposed for use as an indicator showing the presence of pathogenic virus in vaccinated birds. It was proven that pathogenic Newcastle Disease virus persisted in birds from group I for 40 days and 10% of them were in a state of latent infection (clinically healthy). The virus was isolated after method of organ cultures of tracheal explantates. It was established that the HI antibody level in the blood serum of virus carriers was higher as compared with that of the remaining birds in the group. The use of this fact as an indicator for beginning virusological investigations aiming to reveal latent Newcastle Disease infection is proposed.

Animals↗
PubMed1987

Protective effect of antibodies to two viral envelope glycoproteins on lethal infection with Newcastle disease virus.

The protective effect of humoral immunity against lethal infection of chickens with Newcastle disease virus was studied. Chickens hatched from eggs laid by hens vaccinated with live attenuated Newcastle disease virus vaccine possessed antibody to various components of the virus, and were resistant to a challenge with a virulent strain of Newcastle disease virus which was 100 per cent fatal for the offspring of nonvaccinated hens. Passive administration of antiserum raised against whole virions provided susceptible chickens protection comparable to that seen in the birds with maternal antibody. When administered passively, both anti-HN serum with virus neutralizing activity, and anti-F serum with only marginal virus neutralizing activity significantly prolonged the survival of infected birds but failed to achieve the level of protection as afforded by the anti-whole NDV serum. The protection provided by the simultaneous presence of anti-HN and anti-F serum was significantly greater than that afforded by either alone and comparable to that of anti-whole NDV serum, indicating the complementary effect of anti-HN and anti-F antibodies not only in cell cultures as reported previously (19), but also in a natural host.

Animals↗
PubMed1982

Fatty acid modification of Newcastle disease virus glycoproteins.

The fatty acid acylation of Newcastle disease virus hemagglutininin-neuraminidase and fusion glycoproteins was assayed. [3H]palmitate label was associated with cytoplasmic fusion proteins (F0 and F1) and virion-associated F1. In contrast, there was no detectable [3H]palmitate label associated with the hemagglutin-neuraminidase protein in Newcastle disease virus-infected Chinese hamster ovary cells or chicken embryo cells or in virions released from these cells. Thus, fatty acid modification may not be important for the maturation of some glycoproteins.

Acylation↗
PubMed1994

Vaccination of chickens with a recombinant fowlpox virus containing the hemagglutinin-neuraminidase gene of Newcastle disease virus under the control of the fowlpox virus thymidine kinase promoter.

When chickens were vaccinated with a recombinant fowlpox virus (FPV) containing the Newcastle disease virus (NDV) hemagglutinin-neuraminidase (HN) cDNA under the control of the thymidine kinase (TK) promoter and inserted into the FPV TK gene, the FPV antibody response to the recombinant virus was similar to the response to vaccination with standard FPV, and the recombinant virus protected chickens against challenge with virulent FPV. While the presence of the NDV HN cDNA was demonstrated in the recombinant virus, which was stable on serial passage, expression of HN was not detected by hemagglutination, Western blot analysis or immunoprecipitation of infected cell lysate. Chickens vaccinated with the recombinant virus failed to mount an NDV hemagglutination-inhibition antibody response, and they did not resist challenge with velogenic NDV. It was concluded that the TK promoter was too weak to drive the HN gene, but that the insertion into the FPV TK gene did not reduce the immunogenicity of the virus.

Animals↗
PubMed2006

[Construction and characterization of a recombinant fowlpox virus co-expressing F, HN genes of Newcastle disease virus and gB gene of infectious laryngnotracheitis virus].

The Fusion (F) and Haemagglutinin-Neuraminidase (HN) genes of Newcastle disease virus (NDV) and the glycoprotein B (gB) gene of infectious laryngothracheitis virus (ILTV) as well as a LacZ reporter gene were all inserted into a nonessential gene of fowlpox virus (FPV) 017 strain by homologous recombination. The NDV and ILTV genes were each under the control of a fowlpox virus immediate early/late promoter (LP2EP2) while the LacZ reporter gene expression cassette was regulated by a P11 late promoter. A recombinant FPV harboring the F, HN and gB genes as well as the LacZ gene, designated as rFPV-F/HN/gB/LacZ, was obtained after ten cycles of blue plaque purification. The presence of the NDV and ILTV genes was confirmed by PCR. The expression of the recombinant proteins in rFPV-F/HN/gB/LacZ were characterized by Western blot (F and gB proteins) and indirect immunofluorescence test (F, HN and gB proteins). The results demonstrated that all four foreign proteins, which were encoded within a 10 kb gene fragment, could be expressed authentically and efficiently. Compared to the parental virus, rFPV-F/HN/gB/LacZ showed no obvious difference with respect to virus replication and cytopathogenic effects in chicken embryo fibroblasts (CEF) cell culture. Overall, our work suggests that FPV can be a useful live virus vector for the expression of multi- foreign genes against multiple avian pathogens.

Animals↗
PubMed1985

Intracellular processing of the Newcastle disease virus fusion glycoprotein.

The fusion glycoprotein (Fo) of Newcastle disease virus is cleaved at an intracellular site (Nagai et al., Virology 69:523-538, 1976) into F1 and F2. This result was confirmed by comparing the transit time of the fusion protein to the cell surface with the time course of cleavage of Fo. The time required for cleavage of half of the pulse-labeled Fo protein is ca. 40 min faster than the half time of the transit of the fusion protein to the cell surface. To determine the cell compartment in which cleavage occurs, use was made of inhibitors which block glycoprotein migration at specific points and posttranslational modifications known to occur in specific cell membranes. Cleavage of Fo is inhibited by carbonyl cyanide m-chlorophenylhydrazone; thus, cleavage does not occur in the rough endoplasmic reticulum. Monensin blocks the incorporation of Newcastle disease virus glycoproteins into virions and blocks the cleavage of the fusion glycoprotein. However, Fo cannot be radioactively labeled with [3H] fucose, whereas F1 is readily labeled. These results argue that cleavage occurs in the trans Golgi membranes or in a cell compartment occupied by glycoproteins quite soon after their transit through the trans Golgi membranes. The implications of the results presented for the transit times of the fusion protein between subcellular organelles are discussed.

Animals↗
PubMed1987

Reducing agent-sensitive dimerization of the hemagglutinin-neuraminidase glycoprotein of Newcastle disease virus correlates with the presence of cysteine at residue 123.

Viruses within the Newcastle disease virus (NDV) serotype induce a wide array of disease manifestations ranging from an almost apathogenic pattern to the high mortality caused by avirulent or virulent isolates, respectively. A disulfide-linked dimer form of the NDV hemagglutinin-neuraminidase (HN) glycoprotein can be demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under nonreducing conditions for only some of these isolates. For others, indeed the majority of those we have studied, no such reducing agent-sensitive dimeric form of HN is demonstrable. Apparently, there is no causal relationship between disulfide-linked dimeric HN and virulence. Using the deduced amino acid sequence of the dimeric HN of isolate AV as a basis for selection of oligonucleotide primers, we sequenced three additional reducing agent-sensitive dimeric HN glycoproteins and eight for which a disulfide-linked dimer has not been identified, using primer extension and dideoxy sequencing. The deduced amino acid sequences reveal a strict correlation between the presence of cysteine at residue 123 and reducing agent-sensitive dimerization of HN.

Amino Acid Sequence↗
PubMedDate not supplied

Identification of field isolates of infectious bronchitis virus by interference with the La Sota of strain of Newcastle disease virus.

The interference phenomenon of infectious bronchitis virus (IBV) with growth of Newcastle disease virus (NDV) in embryonating chicken eggs (ECE) was used as a diagnostic method. Fifteen field isolates obtained from presumptively infectious-bronchitis-affected chickens were analyzed by the IBV-NDV interference test. Eight isolates were capable of interfering with the growth of the La Sota strain of NDV, as measured by hemagglutination (HA) activity when IBV was inoculated 10 hr before NDV into ECE. The interference was considered specific for IBV, because it could be eliminated by adding homologous anti-IBV serum. The sensibility of this method could be demonstrated, because in some cases low-passage levels of IBV isolates showing HA interference ability were not capable of producing lesions in ECE. Furthermore, serologically negative IBV samples did not interfere with NDV growth. From these results, the IBV-NDV interference test appears to be a potential diagnostic alternative for identifying IBV field isolates.

Animals↗
PubMed2005

Red blood cell elution time of strains of Newcastle disease virus.

Elution time of velogenic, mesogenic and lentogenic strains of Newcastle disease virus was determined. The differences in their elution time were also calculated. Four samples, each of a velogenic strain (VGF2), a mesogenic strain (Komarov) and a lentogenic strain (LaSota) were used for hemagglutination test with 0.6% chicken red blood cells. The time it took for wells of the end hemagglutination points (highest dilution that gave agglutination) to elute was recorded as elution time for each sample. The mean elution time of the three strains of Newcastle disease virus differed significantly (p < 0.05). The velogenic strain gave the highest mean elution time of 118 min, followed by the mesogenic strain with 59 min and the lentogenic strain with 25 min. Based on this result it appears that elution time could form a basis for rough characterization of isolates of Newcastle disease virus into the three major strains.

Animals↗