• BMB Reports
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• v.40 no.5
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• pp.611-616
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• 2007

$E^{rns}$ is an envelope glycoprotein of classical swine fever virus (CSFV) and has an unusual feature of RNase activity. In the present study, we demonstrate that $E^{rns}$ counteracts Newcastle disease virus (NDV)-mediated induction of IFN-$\beta$. For this purpose, $E^{rns}$ fused to the enhanced green fluorescent protein (EGFP) was transiently expressed in porcine kidney 15 (PK15) cells. In luciferase activity assay, $E^{rns}$-EGFP was found to prevent IFN-$\beta$ promoter-driven luciferase expression and block the induction of IFN-$\beta$ promoter mediated by NDV in a dose-dependent manner. Through IFN-specific semi-quantitative RT-PCR detection, obvious decrease of IFN-$\beta$ mRNA in NDV-infected PK15 cells was observed in the presence of $E^{rns}$-EGFP. In contrast, EGFP alone showed none of this block capacity. In addition, $E^{rns}$-EGFP mutations with RNase inactivation were also found to block NDV-mediated induction of IFN-$\beta$. These evidences establish a novel function for CSFV $E^{rns}$ glycoprotein in counteraction of the IFN-$\beta$ induction pathway.

• The Journal of Korean Society of Virology
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• v.28 no.3
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• pp.233-245
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• 1998

The gene encoding F protein of CBP-1 strain, a heat-stable Newcastle disease virus (NDV) isolated from the diseased pheasants in Korea, was characterized by reverse transcription-polymerase chain reaction (RT-PCR), nucleotide and amino acid sequences. Virus RNA was prepared from the chorioallatoic fluid infected with NDV CBP-1 virus and cDNA was amplified by RT-PCR, cloned and sequenced to analyze. The PCR was sensitive as to detect the virus titer above $2^5$ hemagglutination unit. 1.7kb (1,707bp) size of the cDNA was amplified and cloned into BamHI site of pVL1393 Baculo transfer vector. The nucleotide sequences for F protein were determined by dye terminator cyclic sequencing using four pairs of primers, and 553 amino acid sequences were predicted. In comparison of the nucleotide sequence of F gene of CBP-1 with those of other NDV strains, the homology revealed 88.8%, 98.5% and 98.7% with Kyojungwon (KJW), Texas GB and Beaudette C strains, respectively. As the deduced 553 amino acid sequences of F protein of CBP-1 were compared with those of other NDV strains, the homology appeared 89.9%, 98.7% and 98.9% with KJW, Texas GB and Beaudette C strains, respectively. The putative protease cleavage site (112-116) was R-R-Q-K-R, indicating that CBP-1 strain is velogenic type. The amino acid sequences include 6 sites of N-asparagine-linked glycosylation and 13 cysteine residues. These data indicate that the genotype of CBP-1 strain is more closely associated with the strains of Texas GB and Beaudette C than KJW strain.

• Food Science and Preservation
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• v.21 no.5
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• pp.735-739
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• 2014

Ephedra sinica Stapf, known as a medicinal plant, inhibited not only syncytium formation, but also trafficking of viral glycoprotein, hemagglutinin-neuramidase (HN) to the cell-surface. Trafficking of viral glycoprotein to the surface of infected-cells results in syncytium formation in Newcastle disease virus (NDV)-infected baby hamster kidney (BHK) cells. Viral glycoprotein in the infected-cell is processed within the endoplasmic reticulum during routing into surface. The processing of viral glycoprotein like a N-linked oligosaccharide trimming by ${\alpha}$-glucosidase in cell is necessary for virus infection. Methanol extracts showed inhibitory activities ($IC_{50}$ $15{\mu}g/mL$) against ${\alpha}$-glucosidase. This suggested that E. sinica extracts inhibited the cell-surface expression of NDV-HN glycoprotein without significantly affecting HN glycoprotein synthesis in NDV-infected BHK cells.

• Korean Journal of Poultry Science
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• v.27 no.2
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• pp.109-114
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• 2000

The objective of these experiments was to develop an attenuated thermostable Newcastle disease virus(NDV), CBP-1 strain isolated from infected pheasants. Safety, pathogenicity and protective effects against velogenic NDV were also investigated to evaluate if the attenuated NDV, CBP-1 strain could be a candidate for a new NDV vaccine strain. CBP-1 strain was passaged up to the 173 times by nine days old embryonated eggs and chicken embryo fibroblast(CEF) cell cultures. Its sensitivitly to lipid solvents and low pH, thermostability, mean death time(MDT), intracerebral pathogenicity index(ICPI) of one day old chicks and intravenous pathogenicity index(IVPI) of four weeks old chicks were examined. Safety, boosting and protective effects were tested by chicks mortality. CBP-1 NDV strain had significant thermostability at 56$\^{C}$ for 30 minutes. by hemagglutinin activity and egg infectivity test, but was not resistant to lipid solvent. It showed possibility to use as a feed or water vaccine because of the resistance to low pH. MDT, ICPI and IVPI of CBP-1 were attenuated from 51.5, 1.96, 2.60 to 112.4, 1.12, 1.45. These results implied that the 173rd passages in embryonated egg and CEF cell cultures induced a substantial attenuation of the pathogenicity of the parent virus, changing the virulence from velogenic to intermediate between mesogenic and lentogenic. After vaccination with CBP-1 at one day old by drinking water mortality was 17.5%. However, spray vaccination with B1 at one day old, CBP-1 at two weeks ild and challenge with velogenic Kyojeongwon strain at four weeks old showed 93.5% survival rate. Mortality of chicks, vaccination with 173rd passaged CBP-1 strain at one day old, two weeks old and challenge with Kyokeongwon strain at four weeks old, was 20.0%. The results of these studies indicated that partial attenuated CBP-1 strain tended to be a low safety for ND of broiler chicks and would need to be more successive attenuation.

• Journal of Applied Biological Chemistry
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• v.61 no.4
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• pp.379-382
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• 2018

Trafficking process of viral glycoprotein to cell surface results in the syncytium formation when baby hamster kidney (BHK) cells was infected by Newcastle disease virus (NDV). Rhus chinensis gall, well-known as a medicinal plant, inhibited not only syncytium formation, but also trafficking of glycoprotein, hemagglutinin-neuramidase (HN) to the cell-surface. Modification of viral glycoprotein is processed within the endoplasmic reticulum and golgi body during trafficking into surface. R. chinensis gall extracts showed the strong inhibitory activities ($IC_{50}$ $12.5{\mu}g/mL$) against ${\alpha}-glucosidase$, when compared with the ${\beta}-glucosidase$. And this inhibitory activities is increased by the samples in a dose-depedent pattern. These data showed that the extracts of R. chinensis gall inhibited the cell-surface expression of NDV-hemagglutinin-neuramidase glycoprotein without significantly affecting HN glycoprotein synthesis in NDV-infected BHK cells.

• Korean Journal of Veterinary Service
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• v.42 no.3
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• pp.135-144
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• 2019

Low pathogenic avian influenza (LPAI; H9N2) and Newcastle disease (ND) are economically important poultry diseases in Korea. In this study, we investigated pathological features and virus distribution in the lymphoid tissues of chicks experimentally infected with H9N2 and/or ND virus. Six-weeks-old SPF chickens were divided into 4 groups, Control (C), H9N2 (E1), NDV (E2), and H9N2+NDV (E3). E1 group was challenged with 0.1 ml A/Kr/Ck/01310/01 (H9N2) $10^{5.6}$ $EID_{50}$ intranasally, E2 group was challenged with 0.5 ml KJW (NDV) $10^{5.0}{\sim}10^{6.0}$ $ELD_{50}$ intramuscularly, and E3 group was challenged with H9N2, followed 7 days later by NDV. In histopathological examination, E1 group showed depletion and necrosis in bursa of Fabricius, thymus, cecal tonsil, and spleen, whereas E2 and E3 groups were noted severe lymphocyte depletion and necrosis with destruction of lymphoid organs structures. In TUNEL assay, apoptotic bodies were detected in lymphoid organs of all experimental groups, which was most severe in E3 group. H9N2 and ND viruses were predominantly detected in cecal tonsil of E1, E2, and E3 groups by PCR and immunohistochemistry (ICH). In conclusion, co-infection of H9N2 with NDV caused severe pathologic lesions and apoptosis in lymphoid tissues compared to single infections.

• Journal of Veterinary Clinics
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• v.18 no.2
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• pp.152-155
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• 2001

A two-year-old male peacock (Pavo cristatus) showed acute watery green diarrhea, followed by neurological signs including torticollis and muscular tremor. By the hemagglutination inhibition test for detecting the antibody against the Newcastle disease virus (NDV), the peacock serum inhibited the agglutination of chicken red blood cells. Grossly distinctive hemorrhagic lesions were found in the mucosa of proventiculus and intestine and lung. The spleen revealed multiple variable sized necrotic foci. Histologically, the mucosa of gastrointestinal track had hemorrhagic lesions and some of them underwent ulceration. The spleen exhibited multiple variable sized necrotic foci in which fibrin exudation was marked. Central nervous system had mild non-suppulative menin-goencephalitis consisting of vasculitis, perivascular hemorrhage, gliosis and meningitis. The cells particularly in the cerebellum were degenerative to necrotic. Some of these nerve cells revealed characteristic peripheral chromatolysis. From the present serological and pathological findings, it is suggested that NDV causing death of peacock was velogenic viscerotropic strain. This is the first report of the occurrence of velogenic viscerotropic NDV in an adult peacock in Korea.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.12
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• pp.7489-7496
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• 2013

This study was conducted to investigate enhancement of anti-tumor effects of the lentogenic Newcastle disease virus Clone30 strain (NDV rClone30) expressing cytosine deaminase (CD) gene against tumor cells and in murine groin tumor-bearing models. Cytotoxic effects of the rClone30-CD/5-FC on the HepG2 cell line were examined by an MTT method. Anti-tumor activity of rClone30-CD/5-FC was examined in H22 tumor-bearing mice. Compared to the rClone30-CD virus treatment alone, NDV rClone30-CD/5-FC at 0.1 and 1 MOIs exerted significant cytotoxic effects (P<0.05) on HepG2 cells. For treatment of H22 tumor-bearing mice, recombinant NDV was injected together with 5-FC given by either intra-tumor injection or tail vein injection. When 5-FC was administered by intra-tumor injection, survival for the rClone30-CD/5-FC-treated mice was 4/6 for 80 days period vs 1/6, 0/6 and 0/6 for the mice treated with rClone30-CD, 5-FC and saline alone, respectively. When 5-FC was given by tail vein injection, survival for the rClone30-CD/5-FC-treated mice was 3/6 vs 2/6, 0/6 and 0/6 for the mice treated with rClone30-CD, 5-FC or saline alone, respectively. In this study, NDV was used for the first time to deliver the suicide gene for cancer therapy. Incorporation of the CD gene in the lentogenic NDV genome together with 5-FC significantly enhances cell death of HepG2 tumor cells in vitro, decreases tumor volume and increases survival of H22 tumor-bearing mice in vivo.

• Korean Journal of Poultry Science
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• v.37 no.2
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• pp.159-165
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• 2010

Field strains of Ornithobacterium rhinotracheale (OR) were tested on their virulence in specific pathogen free (SPF) embryonated chicken eggs and 3-week-old broilers. When infected with three different OR isolates (OR-161, OR-240 and OR-295) through yolk sac infection route, all strains appeared to be highly pathogenic with responsible mortality 66% and 100% within 12 days post infection (DPI). To test the virulence of OR in the commercial broilers, 3 week-old broilers were grouped depends on the inoculation route of OR isolate (OR-295) through five different infection routes; group 1 (IT: intratracheal), group 2 (IM: intramuscular), group 3 (IV: intravenous), group 4 (aerosol) and group 5 [Mixed: NDV (LaSota)+OR aerosol]. Within 5 to 7 days after inoculation, only broilers given NDV+OR were slightly depressed and coughing, and had mild facial redness. Grossly, foamy and yellow-white yogurt like exudate in the air sacs, predominantly in the abdominal air sacs was present. In histology, infiltration of the air sac epithelium and lamina propria by macrophage and polymorphonuclear granulocytes was seen with cell debris and inflammatory cells, correlated with the presence of OR antigen, as demonstrated by immunohistochemistry. Field strains of OR were able to induce high mortality in the embryonated chicken eggs, whereas broilers were less susceptible to OR infection. Interestingly, in the absence of NDV infection, the four groups of OR single infection only different route showed minimal and temporary microscopic air sac lesions. Thus, Newcastle disease virus (LaSota strain) showed triggering effects on the OR infection in chickens.

• Korean Journal of Veterinary Research
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• v.46 no.3
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• pp.215-224
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• 2006

In order to search the availability of the lectin extracted from Korean mistletoe(Viscum album coloratum) as an adjuvant for the avian vaccines, attempts were made to determine toxicity of the lectin in chicks and its immunostimulating activity on the inactivated vaccines against Newcastle disease virus(NDV). For the determination of toxicity, the lectin was injected into the thigh muscle of SPF chicks(Charles River) of 1-week-old and observed hematologically and pathologically. For the determination of immunostimulating effects, lectin-adjuvanted, inactivated NDV vaccines were injected into the thigh muscle of SPF chicks in the same age group. Sera of the chicks were examined for the hemagglutination-inhibition(HI) antibodies induced, their HI titers and reaction to the NDV antigens. The data were further compared with those from aluminum hydroxide [$Al(OH)_3$]-adjuvanted vaccines and vaccines without adjuvant, and the results are as follows. There were no significant changes observed in the values of RBC, WBC, Hb, PCV, MCV, MCH, MCHC, AST, ALT, BUN, creatinine and total proteins in the chicks administered with lectin of 1.1, 2.2 and $22.2{\mu}g/kg$ body weight, which means the lectin has no effects on blood values and functions of liver and kidney. In histopathologic observation, no lesions were observed in the brain, heart, liver, lung, spleen, kidney, thymus and bursa of Fabricius of the chicks administered with lectin of 1.1, 2.2 and $22.2{\mu}g/kg$ body weight. There were inflammatory lesions, such as congestion, hemorrhage, edema, infiltration of macrophages and coagulation necrosis observed in the thigh muscle of chicks administered with lectin of $22.2{\mu}g/kg$ body weight, whereas no changes were observed in 1.1 and $22.2{\mu}g/kg$ lectin administered chicks. In chicks immunized with lectin($4.4{\mu}g/kg$ of body weight)-adjuvanted B1, LaSota and Ulster 2C vaccines, HI titers in reciprocal values for $log_2$ were 1.8-2.2 at 1 week after vaccination, which was similar with those of 1.5-2.9 by $Al(OH)_3$-adjuvanted vaccines. The HI titers by the lectin-adjuvanted vaccines reached to 3.9-5.3 at 4 weeks, whereas those by the $Al(OH)_3$-adjuvanted vaccines were more high as 7.3-9.3. Meanwhile, the immunostimulating effects of the lectin were recognized while compared to the HI titers with 2.4-3.7 in chicks immunized with vaccines without adjuvants at 4 weeks after vaccination. The chicks immunized with lectin-adjuvanted vaccines were enough to resist challenges by Kyojeongwon strain, a very virulent NDV at 4 weeks after vaccination as well as chicks immunized with $Al(OH)_3$-adjuvanted vaccines. The HI titers by the lectin-adjuvanted vaccines reached to high level as 8.7-10.3 as those with 8.2-9.6 by the $Al(OH)_3$-adjuvanted vaccines at 6 weeks after vaccination, which may be the booster effects by the challenge virus. Antibodies specific to the HN and F antigens of NDV were observed in the sera of both chicks immunized with lectin-adjuvanted vaccines and $Al(OH)_3$-adjuvanted vaccines.