• 한국동물위생학회지
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• 제25권1호
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• pp.87-96
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• 2002

In this study, we wanted to determine if the respirotropic JMK strain of infectious bronchitis virus(IBV), which has a spike glycoprotein gene that is 99% similar to the nephropathogenic Gray strain of IBV, could adapt and cause lesions in the kidney following intracloacal passage in chickens. Two day old specific pathogen free(SPF) cchickens were infected with Gray and JMK strains by the intraocular and cloacal route. Several tissue samples were collected at various times. Viruses were recovered from more tissues and earlier in the infection from chickens infected cloacally than chickens infected intraocularly. Virus was isolated from the kidney of chickens infected with Gray by the intraocular route and JMK by the intracloacal route, but not from chicken given JMK the intraocular route. Histopathologically, interstitial nephritis was observed in Gray infected chickens. However, viral RNA or antigen were not detected in the kidney by in situ hybridization and immunohistochemistry. We further passaged the JMK strain ten times in two day old SPF chickens using cloacal inoculation. We examined the virus titer and histopathological change in the kidney at each passage level. The amount of virus recovered from the kidney was stable throughout this serial passage and the passaged virus did not caused renal damage. Further, virus could not be isolated from the kidney when chickens were infected with the passaged virus by the intraocular route. We conclude that the JMK strain has a strict upper respiratory tract tropism since cloacal passage did not produce nephrotropism or nephropathogenicity.

• 한국동물위생학회지
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• 제31권4호
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• pp.533-546
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• 2008

The efficacy of bovine immune sera to correct the calves with failure of passive transfer(FPT) was evaluated. Immune sera were produced from 14 one-year-old Holstein cattle which were inoculated commercial combined viral vaccine, administered by the challenge of some main enteric or respiratory viruses, aseptically filtered and stored at $4^{\circ}C$ before used. After the treatment of bovine immune sera, Mean transfer factor($mg/d{\ell}$, of IgG administered/kg of body weight) was $5.46{\pm}2.74,\;11.17{\pm}1.27,\;1.40{\pm}0.21$ in K-IP, H-IP and K-IV group, respectively. The corrective effect of bovine immune sera to FPT calf without any clinical signs showed that intravenous route was more effective than intraperitoneal administration(P<0.01). FPT calves with severe signs were not effective response to the immunotherapy used and consequently died within 10 days after the treatment. Ten percentage of controls appeared the clinical signs including diarrhea. On the contrary, there were not any clinical signs in K-IP and H-IV group. There was significant increase of the neutralizing titer against bovine viral diarrhea virus and bovine coronavirus as well as of cell population including CD2, CD4, and monocyte in K-IP and H-IV group after the immunotherapy(P<0.05). Also, K-IP and H-IV group showed the successful correction to FPT within one week after the immunotherapy, but controls had kept the FPT two-four weeks even after the same treatment. Consequently, the results were suggesting that the bovine immune sera could be used the corrective tool to young calves with FPT.

• 대한의생명과학회지
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• 제16권4호
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• pp.229-238
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• 2010

Chicken Mx protein (cMx) induced interferon (IFN) is an antiviral protein to inhibit replication of RNA virus, particularly negative stranded RNA virus, through blockage of transfortation of viral RNA and proteins. In order to determine antiviral effects of cMx from different MHC haplotype chicken, we characterized cMx gene by studying on nucleotide sequencing, antiviral effects to Newcastle disease virus, VSV and MDV, and transcription activities. Three types of eMx genes (2,118 bp) were detected from the different MHC haplotype chickens [B19 (N), B15(F) and B21 (GSP)] chickens, which have showed different susceptible to Marek's disease (MD). Several amino acid substitutions were showed in the cMx. The amino acid 548 and 631 in the cMxs from N and F, chickens susceptible to MD, was Val and Asn which was important on antiviral effects, and showed in resistant cMx. Those in the cMx from GSP, chicken resistant to MD, were same that showed in susceptible cMx. Though every cMx transactivated the expression of the reporter gene, the transcription activation by resistant cMx from N and F was lower compared to that by susceptible cMx from GSP. The decease of the cell growth in the resistant cMx cloned cells was seen in comparison with another cMx clone cells. Replication of NDV and VSV was suppressed in the clones with resistant cMx from N and F. NMx258-transducted cells lack of antiviral effects, and NMx437 or NMx646-transducted cells was showed 60% of antiviral effects compared to NMx705. Mean death time (MDT) and hemaggutination (HA) titer to NDV was long and low in the eggs of N and F lines, but short and high in the egg of GSP line. Interestingly, strong suppression to NDV was observed in the clone with N-Mx and in the eggs of N line. However, the effects of Mx for replication of vvMDV1 have not been. Thus, resistant types of cMx, N- and F-Mx, have showed the anti-viral effects to only RNA virus including NDV and VSV, but not to DNA virus. Antiviral effects of cMx were required whole length of amino acid including Val and Asn in amino acid 548 and 631.

• 한국가금학회지
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• 제32권2호
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• pp.113-123
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• 2005

본 연구는 약병원성 조류인플루엔자 사독백신 개발을 위하여 백신 후보주(ADL0401)로서의 생물학적 특성 및 개발 백신에 대한 면역원성 및 안전성 평가를 실시하였다. 백신 후보주인 ADL0401의 병원성을 조사한 결과 폐사는 없었으며, 임상증상 및 바이러스 재분리율 양상이 국내 표준 야외주인 MS96과 상당히 유사한 생물학적 특성을 나타내어 약병원성 조류인플루엔자의 특징을 갖고 있음을 알 수 있었다. 표준야외주인 MS96과 백신후보주인 ADL0401간의 이종항원의 중화능 실험을 한 결과 r값 = 0.71로 동종항원(r값 = 1)에 비하여 다소 낮은 중화능을 나타내었지만, 두 바이러스간의 항원성엔 큰 차이가 없음을 알 수 있었다. 불활화 능력실험으로 $0.1\%$ Formalin을 $37^{\circ}C$ 조건에서 처리하였을 때 가장 효과적인 결과를 가져왔으며, MS96은 2시간, ADL0401은 1시간만에 다소의 역가 저하는 있었으나 빠르게 불활화가 이루어짐을 확인하였다. 개발 백신의 면역원성 및 안전성 실험 결과 ISA 70 adjuvant 백신 접종시 백신 접종 전후로 폐사율 및 임상증상은 관찰되지 않았으며, 높은 수준의 항체 역가를 형성함으로써 가장 면역원성이 우수한 것으로 확인되었다.

• Asian Pacific Journal of Cancer Prevention
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• 제16권14호
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• pp.5635-5641
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• 2015

Background: Cirrhosis is regarded as a possible end stage of many liver diseases, including viral infection. It occurs when healthy liver tissue becomes damaged and is replaced by scar tissue and finally may lead to hepatocellular carcinoma. Interferons (IFNs)are two general categories, type I and II. Type I includes one beta interferon and over 20 different alpha interferons. Alpha interferons are very similar in how they work, interacting with other proteins on cells like receptors. The main objective of this study was to compare Mx gene productivity post different cell line treatment with imported and Egyptian biosimilar locally produced IFNs, as well as the efficacy of those tested IFNs. Also, an assessment was made of sensitivity of different cell lines as alternatives to that recommended for evaluation of antiviral activity. Materials and Methods: Different cell lines (Vero, MDBK and Wish) were employed to evaluate cytotoxicity using the MTT assay. Antiviral activity was evaluated compared with standard IFN against VSV, Indiana strain -156, on tested rh-IFNs (imported; innovated and Egyptian biosimilar locally produced IFNs) in the pre-treated cell lines previously mentioned. The virus was propagated in the Wish cell line as recommended. Finally we estimated up-regulation of the Mx gene as a biomarker. Results: Data recorded revealed that test IFNs were safe in test cell lines. Viability was around 100%. Locally tested interferon did not realize the international potency limits, while the imported one was accepted compared with the standard IFN. These results were the same either using infectivity titer reduction assay or crystal violet staining of residual non- infected cells. Mx protein production was cell type related and confirmed by the detected Mx gene expressed in imported and locally produced IFN pre-treated cell lines. The expression of the gene was arranged in the order of Vero> wish > MDBK for the imported IFN, while for the Egyptian biosimillar locally produced one it was MDBK> Vero> wish. With regard to the antiviral activity there was a significant difference of imported IFN potency compared with the locally produced IFN (P<0.05), the IFN potential (antiviral activity) was not cell line related and showed non-significant difference for each separate product. Conclusions: Vero cells can be used as an alternative cell line for evaluation of IFN potency in case of unavailable USP recommended cell lines. Alternative potency evaluation assay could be used and proved significant difference in IFN potency in case of local and imported agents. Evaluation of antiviral activity could be used in parallel to viral infectivity reduction assay for better accuracy. Mx gene can be used as a marker for IFN potential.

• 대한수의학회지
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• 제40권4호
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• pp.719-727
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• 2000

A study was conducted to determine the susceptibility of swine to Korean foot-and-mouth disease virus (FMDV; subtype O, isolated from Chungju province) in April, 2ooo. One holstein cow was inoculated intradermolingually with suspension of homogenized tissue from a Korean native cow naturally infected with Korean FMDY. Infected cow was housed with one susceptible cow and one susceptible pig (contact sentinels). Four additional susceptible pigs were housed in the same room but caged separately (non-contacted sentinels). The contacted pig and cow as well as non-contact pigs developed typical clinical signs after 2, 3, and 7 days post exposure, respectively. We compared neutralizing antibody from the animals to FMDV $O_1$ Lombardy, O Taiwan, $O_1$ Campos, and $O_1$ Manisa after 0, 4, 7, 10, 14, 21, 28 days post challenge and post-exposure. The highest viral neutralization titer could be interpreted that serotype O Korea (Chungju isolate) is antigenically more related to $O_1$ Manisa. In addition, immunohistochemistry was used to further characterize Korean FMDV from tissues of infected pigs. Korean FMDV antigen was observed in the tongue, hoof, esophagus, and tonsil tissues of sentinel pigs. These findings suggest that Korean FMD virus isolated from cattle can be rapidly transmitted to pigs both directly and indirectly contrast field observation in which only cattle were clinically ill.

• Biomolecules & Therapeutics
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• 제26권6호
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• pp.576-583
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• 2018

Human rhinoviruses (HRV) are one of the major causes of common cold in humans and are also associated with acute asthma and bronchial illness. Heat-shock protein 90 (Hsp90), a molecular chaperone, is an important host factor for the replication of single-strand RNA viruses. In the current study, we examined the effect of the Hsp90 inhibitor pochonin D, in vitro and in vivo, using a murine model of human rhinovirus type 1B (HRV1B) infection. Our data suggested that Hsp90 inhibition significantly reduced the inflammatory cytokine production and lung damage caused by HRV1B infection. The viral titer was significantly lowered in HRV1B-infected lungs and in Hela cells upon treatment with pochonin D. Infiltration of innate immune cells including granulocytes and monocytes was also reduced in the bronchoalveolar lavage (BAL) by pochonin D treatment after HRV1B infection. Histological analysis of the lung and respiratory tract showed that pochonin D protected the mice from HRV1B infection. Collectively, our results suggest that the Hsp90 inhibitor, pochonin D, could be an attractive antiviral therapeutic for treating HRV infection.

• 대한수의학회지
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• 제52권4호
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• pp.223-230
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• 2012

The worldwide distribution and continuing genetic mutation of avian influenza virus (AIV) has been posed a great threat to human and animal health. A comparison of 3 isolates of AIV H9N2, A/chicken/Korea/KBNP-0028/00 (H9N2) (KBNP-0028), A/chicken/Korea/SNU8011/08 (H9N2) (SNU 8011) and an inactivated oil vaccine strain A/chicken/Korea/01310/01 (H9N2) (01310), was performed. The former 2 AIVs were isolated from field cases before and after the application of an inactivated H9N2 vaccine in 2007, respectively. The antigenic relationship, viral shedding, tissue tropism and genetic analysis were examined. The comparison of virus shedding from the cloaca and the oropharynx revealed that both isolates were more frequently isolated from the upper respiratory tract (90~100%) 1 day post inoculation (DPI) compared with isolation 5 DPI from gastrointestinal tracts (10~60%). Moreover, the isolate KBNP-0028 were recovered from all organs including bone marrow, brain and kidneys, indicating higher ability for broad tissue dissemination than that of SNU 8011. KBNP-0028 replicated earlier than other strains and with a higher titer than SNU 8011. In full-length nucleotide sequences of the NA gene and a partial sequence of the HA gene of SNU 8011, we found that there might be significant changes in tissue tropism, virus replication and genetic mutation in AIV H9N2 isolates.

• KSBB Journal
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• 제16권6호
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• pp.579-591
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• 2001

The cell growth inhibitor effect of cervical cancer cells was investigated by liposome mediated transfection (pRcCMVp53/lipofectin) and by transfection using adenovirus (AdCMVp57). The papilloma virus cancer cell lines we used in this study were HPV16 positive, having inhibiter gene, wild p53 gene, CaSki, SiHa, HPV18 positive HeLa, HeLaS3 and HPV negative C33A, HT3. LacZ gene of E.coli was used as the marker gene for the transfection efficiency. The effect on the inhibition of tumor cell growth was measured by cell count and cell viability though ELISA analysis and MTT assay. The inhibition of tumor cell growth was confirmed by measuring each assay for six days, comparing with the normal control cell growth. The cell growth of cervical cancer calls by transfection was significantly reduced and showed tittle differences among the cell lines. To eliminate the potential problem of Ad(adenovirus) contamination during rAAV production, rAAV can be produced by a triple transfection of vector plasmic, packaging plasmid, and adenovirus helper plasmid. To examine the helper functions of Ad plasmids on the production of rAAV vector, we carried out cotransfection of three plasmids, AAV vector, packaging construct, and Ad helper plasmids. The optimized transfection condition for calcium phosphate method is 25ug of total DNA per 10-cm-diameter plate of 293 cell. We found that rAAV yields peaked at 48hr after Ad infection. The titer of rAAV was measured by the dot blot analysis to measure the number of particles/ml based on the quantification of viral DNA. Recent1y, Kombucha(fungi) was identified as a very potent antileukefic agent. In the present study, effect of natural toxin(plankton) and Kombucha is PSP(GTXI-3, neoSTX), on various MTT assay cervical cancer cell line. Toxin(GTX 1-3, neoSTX) also inhibited the proliferation in primary cervical cancer calls in a dose-dependent toxin concentration. These results showed that toxin was very potent in inhibiting the proliferation of cervical cancer calls in vitro. Toxins and Kombuoha exhibited a dose dependent inhibition of cellular proliferation in cancer cell line.

• 한국임상수의학회지
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• 제22권4호
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• pp.365-370
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• 2005

Enzootic pneumonia caused by Mycoplasma hyopneumoniae is responsible for major economic losses in pig herds of world wide. Mycoplasma hyopneumoniae can also act as a primary pathogen of porcine respiratory disease complex followed by bacterial or viral infection. This study was carried out to investigate the prevalence of mycoplasmal pneumonia of slaughtered pigs in Jeju for two years. The lungs and sera of 214 cases were examined for gross and microscopic lesions of the lungs, immunohistichemistry test for Mycoplasma hyopneumoniae antigen and enzyme-linked immunohistichemistry assay (ELISA) for serum antibody titer. Pulmonary consolidation was observed in the lungs of 163 pigs $(76.1\%)$ with average gross lesion score of $6.0\%$., Bronchointerstitial pneumonia was most frequently observed $(78.5\%)$. The incidence of pulmonary consolidation was decreased in vaccinated pigs compared to that of non-vaccinated pigs. The rate of consolidation in the lungs was significantly decreased in the vaccinated pigs (P<0.05). Antigen of Mycoplasma hyopneumoniae was identified by immunohistichemistry test in the lungs of 174 pigs $(81.3\%)$. ELISh antibodies to Mycoplasma hyopneumoniae were detected in 154 pigs $(72.0\%)$. These results showed the prevalence of swine pneumonia and the incidence of Mycoplasma hyopneumoniae in slaughtered pigs of Jeiu province. We expect that these results would be helpful for the control of swine mycoplasmal pneumonia and porcine respiratory disease complex in Jeju.