• 대한수의학회지
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• 제41권3호
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• pp.343-350
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• 2001

The neutralization epitopes of the outer capsid protein VP4 of a porcine rotavirus, Gottfried strain, were studied using neutralizing monocolonal antibodies(N-MAbs). Eight N-MAbs which are specific for the VP4 of Gottfried strain were used for analyzing the antigenic sites of VP4. Three different approaches were used for this analysis; i)testing the serological reactivity of each N-MAb against different G and P types of human and animal rotavirusese ii) analyzing N-MAb-resistant viral escape mutants and iii) performing nucleotide sequence analysis of the VP4 gene of each N-MAb-resistant viral escape mutant. From experimental results, at least four antigenic sites(I, II, III, and IV) were identified. Antigenic site I recognized by N-MAbs 24B9, 23G10, and 26A2 was separated from antigenic site II recognized by N-MAbs 30H5, 32B3, and 29B3. However, these antigenic sites were overlapped with antigenic site III recognized by N-MAb 21A1. The other antigenic site IV recognized by N-MAb 16D2 was separated from antigenic sites I, II, and III.

• 대한수의학회지
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• 제38권2호
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• pp.319-327
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• 1998

The nucleotide sequences of spike (S) glycoprotein containing antigenic sites A and D of TGEV isolated in Korea were determined and compared with published sequences for TGEVs. The TGEV 133 and DAE5 strains had 97.40% nucleotide sequence similarity. The overall nucleotide sequence similarity of the 133 and DAE5 strains compared with other TGEV strains was between 96.86% and 99.15%. The similarity of the predicted amino acid sequence of the 133 and DAE5 strains was 94.93%. The TGEV 133 and DAE5 strains had 94.93-98.61% amino acid similarity with published sequences of other TGEV strains. The sequences of amino acid codons in the antigenic sites A and D were identical among all the viruses although there were several nucleotide changes in region containing antigenic sites A and D of Korean TGEV isolates. By phylogenetic analysis of the sequences, two Korean isolates 133 and DAE5 seemed to be derived from different lineages. These studies showed that a distinct difference in genome exists among TGEV field strains isolated in Korea.

• Parasites, Hosts and Diseases
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• 제32권2호
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• pp.111-116
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• 1994

우리나라 소에 유행하고 있는 Theileria sergenti의 예방을 위한 연구의 일환으로 긴sergeki merozoite 수용성 항원중에서 면역원성 물질로 보고한 바 있는 34. 29 및 18 kDa와 항원성 물질인 45 kDa에 대하여 Peptdie 구성 특성을 밝히기 위하여. 이들 물질에 대한 아미노산 서열을 결정하였다. 이를 생합성하여 Chou-Fasmanprediction법에 의해 항원 결정기를 확인하였던 바. 45 kDa. 34 kDa. 29 kDa 그리고 18 kDa의 Polypeptide는 6 4 2 그리고 0 개의 항원 결정기를 갖고 있었다. 그런데 45 kDa 물질은 항 T. serpenti혈청에 대하여 면역원성이 인정되않았던 물질이었다.

• 대한수의학회지
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• 제38권1호
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• pp.53-63
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• 1998

Total of 1085 swine sera (1996-1997) from nation-wide were tested for the presence of antibodies to influenza A virus. Fifty nine percent of the tested sera showed seropositive by HI test. Positive sera consisted of 24--- of H3, 15--- of H1, and 20--- of the sample had both antibodies, respectively. Sera collected from various region represented 7~27--- seropositivity to H1N1, 15~25--- to H3N2, respectively. Swine influenza field isolate from nasal swab was characterized antigenically and genetically to elucidate its relatedness with other known strains of influenza A virus. The study was focused on the HA gene which is related to pathogenecity and antigenic variability of the influenza virus. By RT-PCR using influenza A/H1N1 specific primers, influenza virus H1N1 specific DNA fragment was amplified from A/Swine/Iowa/15/30(H1N1), US field isolate but not in H3N2 strain. PCR products were sequenced by dideoxy chain termination method to determine nucleotide homology with other strains of influenza A virus. The US field isolate and A/Swine/Indiana/1726/88 strain had 97--- of nucleotide homology and 98--- of amino acid homology. Based on the results obtained from this experiment, the field isolate was genetically related to A/Swine/Indiana/1726/88 and had higher homology with A/Swine/Indiana/1726/88 than with classical swine influenza virus, A/Swine/Iowa/15/30. The field isolate had no amino acid changes at the antigenic site compare to that of the A/Swine/Indiana/1726/88. The proteolytic enzyme cleavage site between HA1 and HA2 had no alteration and the amino acid arginine was intact. There is no evidence has been found that the field isolate has genetic shift or genetic drift which might altered antigenic determinant.

• Journal of Veterinary Science
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• 제23권4호
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• pp.55.1-55.12
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• 2022

Background: ASF was first reported in Kenya in 1910 in 1921. In China, ASF spread to 31 provinces including Henan and Jiangsu within six months after it was first reported on August 3, 2018. The epidemic almost affected the whole China, causing direct economic losses of tens of billions of yuan. Cause great loss to our pig industry. As ELISA is cheap and easy to operate, OIE regards it as the preferred serological method for ASF detection. P54 protein has good antigenicity and is an ideal antigen for detection. Objective: To identify a conservative site in the African swine fever virus (ASFV) p54 protein and perform a Cloth-enzyme-linked immunosorbent assay (ELISA) for detecting the ASFV antibody in order to reduce risks posed by using the live virus in diagnostic assays. Method: We used bioinformatics methods to predict the antigen epitope of the ASFV p54 protein in combination with the antigenic index and artificially synthesized the predicted antigen epitope peptides. Using ASFV-positive serum and specific monoclonal antibodies (mAbs), we performed indirect ELISA and blocking ELISA to verify the immunological properties of the predicted epitope polypeptide. Results: The results of our prediction revealed that the possible antigen epitope regions were A23-29, A36-45, A72-94, A114-120, A124-130, and A137-150. The indirect ELISA showed that the peptides A23-29, A36-45, A72-94, A114-120, and A137-150 have good antigenicity. Moreover, the A36-45 polypeptide can react specifically with the mAb secreted by hybridoma cells, and its binding site contains a minimum number of essential amino acids in the sequence 37DIQFINPY44. Conclusions: Our study confirmed a conservative antigenic site in the ASFV p54 protein and its amino acid sequence. A competitive ELISA method for detecting ASFV antibodies was established based on recombinant p54 and matching mAb. Moreover, testing the protein sequence alignment verified that the method can theoretically detect antibodies produced by pigs affected by nearly all ASFVs worldwide.

• 대한바이러스학회지
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• 제28권2호
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• pp.139-146
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• 1998

Recombinant polioviruses have been developed by many research groups for use as vaccine vector because poliovirus induces mucosal immunity as well as humoral immunity through oral uptake. We assessed the potential use of poliovirus as a T-cell epitope carrier. Recombinant poliovirus V129 5L was constructed to have a substituted T-helper epitope from the core protein of Hepatitis B virus at neutralization antigenic site 1 on its VP1 capsid protein. The recombinant virus replicated less efficiently than type 1 poliovirus Mahoney strain. The V129 5L formed a little smaller plaques than the Mahoney strain and showed some 1.25 log unit lower titer at the peak in the one-step growth kinetics though it had similar growth profile to that of the Mahoney strain. Since V129 5L recombinant virus was genetically stable even after 24 successive passages in HeLa cells, the antigenic site 1 on VP1 capsid protein was confirmed for its ability of carrying T cell epitope. The genetic stability of V129 5L also indicated that recombinant poliovirus can be successfully utilized for the development of the multivalent vaccines.

• 한국동물학회지
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• 제22권3호
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• pp.115-124
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• 1979

경골어류 드렁허리와 가물치에서 젖산수소이탈효소의 세가지 동질사량체를 순수 정제하였다. 이들에 대한 항혈청을 얻어 젖산수소이탈효소와 면역화학적 반응을 실시하였다. 하부단위체의 활성자리는 항원결정군에 속해 있지 않으며, 항혈청내의 항체 또는 미지의 물질이 하부단위체의 하전을 변화시키며, 두 하부단위체는 공통의 항원결정군을 갖고 있음이 확인되었다.

• 한국연초학회지
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• 제6권2호
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• pp.141-146
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• 1984

Two PVY strains (PVY-VB and PVY-VN) isolated from tobacco in Korea were compared for their serological relationship with other 8 strains which were obtained from tobacco or potato in different countries. One of these strains, PVY-Argentina showed the spur reaction to PVY-VB and PVY-VN antisera in SDS-agar gel double diffusion plates. The two Korean PVY strains were closely related to other strains except for one, PVY-Argentina when antigen-antibody reciprocal absorption tests were conducted. It is suggested that the strain, PVY-Argentina, is a new serotype containing a specific antigenic site different from other 9 strains tested.

• 한국미생물·생명공학회지
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• 제20권3호
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• pp.301-310
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• 1992

요로계 감염에 대처함에 있어 hemolysin과 Gal-Gal pili의 multicomponent 백신 개발 가능성을 조사하기 위하여 본 연구를 수행하였다. 요로계 감염환자로부터 분리되어 그의 hemolysin gene 의 염기배열과 아미노산 배열이 밝혀진 대장균 J96 균중의 hemolysin에 대하여 10곳에 해당하는 20 mer-oligonucleotide를 합성하였다. 이들 probe를 사용하여 요로계감염 환자들로부터 분리한 wild type 대장균들의 DNA에 대하여 hemolysin gene 의 상동성을 조사하였는 바 8개의 probe는 거의 모든 hemolysin signal을 보여주었고 HA484는 28.3, HA661은 71.7의 positive signal을 나타내었다.이는 요로계 감염 대장균들의 Hemolysin gene의 상동성이 매우 높은 것을 의미한다. 또한 J96 hemolysin에 대한 12개의 MAB은 모두 wild type 대장균들이 분비하는 90% 이상의 균주에 대하여 양성 immunoblotting을 나타내었다. 특히 J96 hemolysin의 functional site를 가장 강력하게 block할 수 있는 monoclonal antibody MAＢ132가 모든 wild type으로부터 분리된 hemolysin의 function을 중화시킬수 있는 결과는 J96의 cloned hemolysin product를 vaccine으로 사용할 수 있는 가능성을 증대시키는 결과라고 할 수 있다.

• Parasites, Hosts and Diseases
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• 제35권1호
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• pp.31-38
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• 1997

BALB/c 마우스에서 감염부위와 감염기간에 따른 숙주 체액 면역반응의 변화를 알아보고자 하였다. 배양한 큰리슈만편모충의 전편모형(promastigote)을 BALB/c 마우스의 코. 등 발바닥 부위로 나누어 각각 $3{\;}{\times}{\;}10^6$마리씩 피하 감염 후 10-100일 동안에 궤양의 형성과정을 관찰하고 채혈하여 SDS-PAGE와 면역이적법을 시행하여 각 부위별로 나타나는 항체 반응을 관찰하였다. 외관상으로는 감염 15일부터 코에 감염시킨 마우스에서 먼저 궤양이 형성되기 시작하였고. 코에 궤양이 나타난 후 2-3일 뒤에 발에서 궤양이 형성되었으며 등에서는 감염시킨 후 90일이 되어서야 궤양이 관찰되었다. 감염후 20일에 실시한 면역이적법에 의하면 코 감염군에서는 202, 139, 98, 83, 81, 67, 65, 62, 59, 54, 52, 42, 26, 23 kDa의 항원성 분획이 관찰되었고 발 감염군에서의 항원 분획양상도 코 감염군과 같았으나 등감염군에서는 202, 83, 81, 65 kDa의 희미한 항원성 분획이 관찰되었다. 그러나 감염 후 90일이 경과한 등 감염군에서는 202, 83, 81, 74, 67, 65, 62, 59, 54, 52, 20, 17 kDa의 항원 분획이 관찰되었다 이상의 결과로부터 감염부위와 감염기간에 따라 큰리슈만편모충에 대한 혈청반응이 항원 분획에 따라 다르게 나타남을 관찰하였다. 이 차이는 세 감염부위의 온도차에 의한 결과일 가능성도 있으나 다른 부위에 감염될 경우 한 숙주 내에서도 다른 면역반응이 유발되어 나타날 수도 있다고 추측하였다. 특히 궤양 형성 시기와 혈청 내 67-52 kDa 분획에 대한 항체 출현 시기가 일치하는 것으로 보아 궤양 형성에 이 항체가 관여할 가능성이 있음을 시사한다.