• 대한바이러스학회지
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• 제27권2호
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• pp.151-160
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• 1997

In view of the potential of replicase protein as a diagnostic reagent for human caliciviruses (HuCVs), we have cloned and over-expressed this gene from the Snow Mountain-like Korean strains in Escherichia coli as a fusion protein with glutathione S-transferase (GST), and described the preliminary antigenic characterization of the recombinant products. Each 470bp fragment corresponding to highly conserved region of RNA-dependent RNA polymerase was generated by RT-PCR from stools of two diarrheal children, cloned in pMOSBlue T-vector, and subcloned between the EcoRI and SalI restriction sites of pGEX-4T-3, a GST gene fusion vector, yielding $pGCV_{pol}$. This construct expressed a Snow Mountain-like HuCV replicase under the control of the IPTG-inducible tac promoter. An extract prepared by sonication of the E. coli cell inclusion bodies bearing $pGCV_{pol}$ products was purified and analyzed by SDS-PAGE. After Coomassie blue staining, it was shown that the recombinant replicase migrated on the gels with an approximate molecular mass of 46.5 kDa, that was subsequently cleaved into a 26 kDa GST fragment and a 20.5 kDa replicase protein upon digestion with thrombin protease. The replicase was recognized on immunoblotting with the sera from symptomatic children with the HuCV-associated diarrhea but not by asymptomatic sera from adults. The results presented the first biological activity of individually expressed HuCV replicase subunit and provided important reagents for diagnosis of HuCV infection.

• Biomolecules & Therapeutics
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• 제4권1호
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• pp.1-6
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• 1996

In the present study, the antigenic potential of G009, a polysaccharide isolated from Ganoderma lucidum IY009, was determined in BALB/C mice and Hartley guinea pigs. Antigenicity tests, including passive cutaneous anaphylaxis (PCA), active systemic anaphylaxis (ASA) and indirect hemagglutination test (IHA) were performed according to the established guidelines of National Institute of Safety Research. The results were as follows: 1. Mice showed no production of antibodies against G009 sensitized with an adjuvant, aluminum hydroxide gel (alum), when judged by the heterologous PCA test in rats. Meanwhile, antibodies against ovalbumin (OVA) sensitized with alum were clearly detected. 2. In the studies with guinea pigs, both the sensitization of G009 alone and of G009 with complete Freund's adjutant (CFA) did not produce positive reactions in homologous PCA. In the case of ASA, however, G009 alone and G009 with CFA produced positive reactions. 3. No G009 specific reaction was observed in an IHA assay using sera isolated from G009 sensitized mice. These findings suggest that G009 have no antigenicity potential in mice but may have weak antigenicity in guinea pjgs.

• 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.

• 대한수의학회지
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• 제42권2호
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• pp.253-260
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• 2002

Fowl red mite, Dermanyssus gallinae, is the most important ectoparasite affecting egg layers worldwide. More than 35 compounds have been used for fowl red mite control. Although some of them are efficient, several compounds are unsuitable in terms of food safety and environmental problems. Some compounds are efficient in theory but inadequate in practice. It is also expensive in material and labor to control effectively. Effective doses are very close to toxic doses and repeated treatment is required. Repeated, long term treatment of compounds on fowl red mite populations, may cause heritable resistance against the mites. In this study, antigenicity of fowl red mite and house dust mite, Dermatophagoides pteronyssinus, were identified by SDS-PAGE, silver staining, Western blotting and ELISA to investigate immune effects against fowl red mite using somatic antigens of fowl red mite and house dust mite. By SDS-PAGE, silver staining and Western blotting, several common antigens (110, 60, 56, 49, 46 kDa) of both fowl red mite and house dust mite were recognized. To identify immune effect of somatic antigens of fowl red mite and house dust mite, sixty white leghorn broilers(1 week old) were used. Among sixty white leghorn broilers, twenty were immunized with fowl red mite somatic antigens(Group I), twenty immunized with house dust mite antigens(Group II), and twenty were control group without antigen(Group III), respectively. After immunization, it was identified that antibody titers were increased both in group and II. Then all groups were challenged with fowl red mites. After 2 months, measurements of body weights, packed cell volume(PCV), ELISA OD values and numbers of mites were significant(p<0.05). These results suggest that fowl red mite and house dust mite, which are easy to collect and maintain, can be good vaccine candidates against fowl red mite in chicken.

• 대한수의학회지
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• 제42권2호
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• pp.209-217
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• 2002

This study was performed to produce monoclonal antibodies (mAb) specifically reacting with chicken leukocyte surface antigens. Popliteal lymph node cells of BALB/c mice previously immunized through foot-pad with peripheral blood mononuclear cells (PBMC) of chickens separated by Ficoll-Histopaque method. They were fused with P3X63Ag14 mouse myeloma cells. A total of 34 hybridomas secreted antibodies specifically binding to the PBMC. According to the reactivity patterns with PBMC, the mAbs were divided into 4 groups. Group 1 mAbs (IIB3, IIB10, IIE10) specifically reacted with non-adherent lymphocytes but not with adherent cells which were mainly composed of thrombocytes and monocytes in PBMC culture. These mAbs were reactive with 25-59% of thymus cells and 42-64% of spleen cells of chickens. They did not show any significant reactivity with cells in the bursa of Fabricius, T-cell (MDCC-MSB1) and B-cell (LSCC-1104B1) lines. These results indicate that Group I mAbs specifically reacted with T-lymphocyte subpopulation. Monoclonal antibodies in Group II (IC6, IG2-2 and IID9) showed specific reactivity with monocytes but not with thrombocytes or non-adherent cells in PBMC culture. These mAbs, though not reacted with the chicken macrophage cell line, HD11, also bound to macrophages of the spleen and lung in immunohistochemical staining. Five mAbs in Group III showed characteristics of binding to lymphocytes and monocytes, but not to thrombocytes. Twenty-three mAbs in Group IV showed specific reactivity to lymphocytes, monocytes, and thrombocytes. Two mAbs (IC3 and IE9) in Group IV reacted with most of PBMC.

• 대한수의학회지
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• 제44권1호
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• pp.65-71
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• 2004

This study was carried out to produce IgY against B. bronchiseptica, parvovirus and distemper virus that are major pathogens in alimentary and/or respiratory diseases of dogs. In the comparison of adjuvants, ISA70 was the best in the rapid induction and maintence of antibody titers. Agglutination antibody titers against B. bronchiseptica were 1:1,280 ~ 1:10,240 in sera and 1:160 ~ 1:1,280 in egg yolk. Hemagglutination inhibition(HI) titers against parvovirus in sera and egg yolk were 1:80 ~ 1:320 and 1:64 ~ 1:256, respectively. Virus neutralization titers against canine distemper was 1:8 ~ 1:64 in sera and egg yolk. These results suggested that egg yolk antibody titers could be variable according to a sort of adjuvant and antigens of the pathogens.

• 대한수의학회지
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• 제44권1호
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• pp.57-64
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• 2004

The common pathogen Salmonella enteirtidis (S. enteritidis) is the major cause of foodborne disease. Protein identification by peptide mass fingerprinting using the matrix-assisted laser desorption ionization time of fight (MALDI-TOF) mass spectrometry (MS) can analysis unambiguously identity the spots from 2-dimensional electrophoresis (2-DE) gel. In this report, we examined protein components from patterns of S. enteritidis proteins. In addition, antigens that are recognized by sera can be identified by immunoblotting. This study that 2-DE analysis of S. enteritidis yields useful information concerning S. enteritidis proteome, the results that have been obtained led to a more detailed understanding of Salmonella pathology and open further interesting fields for future work.

• 대한수의학회지
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• 제55권2호
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• pp.89-95
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• 2015

Johne's disease, caused by Mycobacterium avium subsp. paratuberculosis (MAP), is one of the most widespread and economically important diseases in cattle. Current diagnostic methods are based on the detection of anti-MAP antibodies in serum or isolation of the causative agent. However, these techniques are often not applicable for cases of subclinical infection due to relatively low sensitivity. Therefore, finding new antigen candidates that strongly react with the host immune system had been attempted. To effectively detect infection during the subclinical stage, several antigen candidates were selected based on previous researches. Characteristics of the selected antigen candidates were analyzed using bioinformatics-based prediction tools. A total of nine antigens were selected (MAP0862, MAP3817c, MAP2077c, MAP0860c, MAP3954, MAP3155c, MAP1204, MAP1087, and MAP2963c) that have MAP-specific and/or high immune responses to infected animals. Using a transmembrane prediction tool, five of the nine antigen candidates were predicted to be membrane protein (MAP3817c, MAP3954, MAP3155c, MAP1087, and MAP1204). Some of the predicted protein structures identified using the I-TASSER server shared similarities with known proteins found in the Protein Data Bank database (MAP0862, MAP1204, and MAP2077c). In future studies, the characteristics and diagnostic efficiency of the selected antigen candidates will be evaluated.

• Journal of Chest Surgery
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• 제34권1호
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• pp.64-71
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• 2001

배경: 최근 치료법의 진보에도 불구하고 진행성 식도암의 예후는 5년 생존율이 10% 이하로매우 불량하기 때문에 식도암에 대한 새로운 치료방법의 하나로 암면역 치료가 대두되고 있다. 암면역 치료를 위해서 MAGE 등 종양 특이항원이 연구의 대상이 되고 있으나 국내에서는 아직 이에 대한 연구가 없다. 대상 및 방법: 1995년 1월부터 1998년 12월까지 고신대학교 복음병원 흉부외과에서 수술 치험한 125례의 식도암중 병리조직 보관상태가 양호한 편평세포암 79례를 병기에 따라(1병기 19례, IIa병기 19례, IIb병기 10례, III병기 21례, IV병기 10례) 무작위로 추출하고 대조군으로 평활근종 20례와 정상 식도점막 20례를 대조군으로 하여 DO7 단클론 항체와 항 MAGE-3 단클론 항체 57B를 이용하여 면역조직화학검사를 시행하여 변형 p53 단백과 MAGE-3 유전자 산물의 발현율을 조사하고 식도암 조직에서 질병의 진행도를 반영하는 병기에 따른 발현율 및 변형 p53 단백과 MAGE-3 유전자 산물의 발현율간의 상관관계를 조사하였다. 결과: 식도암조직에서 변형 p53 단백과 MAGE-3 유전자 산물의 발현율은 각각 51.9%, 60.8%의 발현율을 보였으나 식도평활근종과 정상 식도점막에서는 한례도 발현되지 않아 변형 p53 단백과 MAGE-3 유전자 산물은 대조군에 비해 식도암 조직에서 의미있게 발현되었다(p<0.001). 변형 p53 단백과 MAGE-3 유전자 산물의 발현은 I병기에서 68.4%, 52.6%, IIa병기에서 57.9%, 47.6%, IIb병기에서 60%, 70%, III병기에서 33.3%, 71.4%, IV병기에서 40%, 70% 각각 발현되어 병기에 따른 발현율의 차이는 없었다(p=0.193, p=0.452). 식도암 조직내에서 변형 p53 단백과 MAGE-3 유전자 산물의 발현간에는 상관관계가 없는 것으로 나타났다(p=0.679). 결론: 이상의 결과로 변형 p53 단백과 MAGE-3 유전자 산물의 발현은 식도암에서 예후인자로서의 역할은 할수 없으나 식도 편평세포 암조직에서만 특이하게 높은 빈도로 발현됨으로써 식도암도 면역치료의 대상이 될 수 있음을 확인하였다.

• 대한미생물학회지
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• 제16권1호
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• pp.39-48
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• 1981

보체결합반응은 감염초기의 항체검출에 대한 민감성이 높기 때문에 사람과 동물의 만성질환, 특히 매독, 브루셀라병. 결핵병, �냑鵝뵉퓌鑿�, 클래미디아감염증, 마이코플라스마감염증, 콕시오이드병, 히스토플라스마병 등의 면역학적진단수기와 예방관리를 위한 전염병학적연구목적에 흔히 사용된다. 그러나 검사혈청이 항보체성 또는 친보체성을 갖는 경우는 특이항체를 검출할 수 없는 단점을 갖고 있다. 그러므로 이 연구에서는 친보체성분을 갖는 돼지혈청을 항보체성 사람, 토끼 및 소혈청에 보강하여 특이 항체검출을 위한 민감성과 특이성에 관한 기초실험을 실시하였다. 결핵에 감염된 사람, 토끼 및 소혈청중에 높은 항보체작용이 있는 혈청에 친보체성 돼지혈청을 보강할 때 결핵균(다당체 및 단백질항원)에 대한 특이항체가 검출되었다.