• 한국가금학회지
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• 제15권3호
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• pp.207-210
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• 1988

뉴켓슬병바이러스(NDV)인 LaSota 주를 SPF 발육난의 요막강내에 증식시켜 순수 정제한 것을 BALB/c 흰쥐에 면역시킨 후 추출한 비강세포와 흰쥐 골수암세포와의 융합방법에 의하여 NDV에 특이하게 작용하는 단크론성항체(MCA)를 생산하는 3주의 Hybridoma틀 작성하였다. 이 3주의 MCA는 모두 IgG형에 속하였으며 흰쥐 복강 내에 접종하여 생산된 복수항체의 항체가는 간접형광항체법으로 $10^3$-$10^6$에 달하였고 약독 및 강독 NDV에 모두 동일한 수준으로 작용하였다. 중화능은 인정되지 않았고 3주중 1주만이 별구응집 억제능을 약하게 나타냈다. 이 MCA를 이용하여 간접형광항체법으로 인공 감염시킨 닭에서 NDV항원 검출을 시도한 결과 기관점막을 비롯한 각종 장기의 도말표본에서 접종 3일 후부터 뚜렷한 검출이 가능하였다.

• Journal of Microbiology and Biotechnology
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• 제24권12호
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• pp.1719-1727
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• 2014

In the present study, whispovirus immediate early 1 promoter (ie-1) was used to initiate surface expression of the hemagglutinin (HA) protein of Egyptian H5N1 avian influenza virus (AIV) by using the baculovirus expression vector system. The HA gene and whispovirus ie-1 promoter sequence were synthesized as a fused expression cassette (ie1-HA) and successfully cloned into the pFastBac-1 transfer vector. The recombinant vector was transformed into DH10Bac competent cells, and the recombinant bacmid was generated via site-specific transposition. The recombinant bacmid was used for transfection of Spodoptera frugiperda (Sf-9) insect cells to construct the recombinant baculovirus and to induce expression of the HA protein of H5N1 AIV. The recombinant glycoprotein expressed in Sf-9 cells showed hemadsorption activity. Hemagglutination activity was also detected in both extra- and intracellular recombinant HAs. Both the HA and hemadsorption activities were inhibited by reference polyclonal anti-H5 sera. Significant expression of the recombinant protein was observed on the surface of infected insect cells by using immunofluorescence. SDS-PAGE analysis of the expressed protein revealed the presence of a visually distinguishable band of ~63 kDa in size, which was absent in the non-infected cell control. Western blot analysis confirmed that the distinct 63 kDa band corresponded to the recombinant HA glycoprotein of H5N1 AIV. This study reports the successful expression of the HA protein of H5N1 AIV. The expressed protein was displayed on the plasma membrane of infected insect cells under the control of whispovirus ie-1 promoter by using the baculovirus expression vector system.

• 생명과학회지
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• 제14권1호
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• pp.148-153
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• 2004

가자 추출물을 취하여 사람, 돼지, 쥐 및 개 등의 3% 적혈구용액으로 응집력시험을 행한 결과 7가지 적혈구 모두에 렉틴 활성이 나타났으며 $LD_{50}$는 390 mg/kg으로 측정되었다. 생리 활성에 대한 영향을 알아보고자 생체 내외인성 요인에 의한 친전자성 물질로 생체내에서 독작용, 노화, 발암 및 면역 억제작용을 유발하는 원인 물질인 free radical생성에 관여하는 효소인 XO 및 AO의 활성을 측정 한 결과, XO 에서는 일주일간 매일 300 mg/kg의 용량으로 가자 추출물을 투여한 군이 정상군보다 5배 증가되는 결과를 나타내었고 AO에서는 정상군보다 시료 투여군이 2배 증가되는 결과를 나타내었다 glutathione은 단백질이나 DNA합성, amino acid의 이동 반응 및 thiol기의 저장 등과 같은 생물학적으로 중요한 여러 가지 반응에 직접 관여하는 물질이다[16〕. 이에 간장 조직의 glutathione농도를 측정한 결과 간장 독성의 유발로 인한 효소 활성은 정상군에 비하여 1주간 매일 300 mg/kg의 용량으로 시료 투여한 군의 효소 활성이 79% 감소됨을 볼 수 있었다. 체내의 여러 가지 해독반응과정에 관여하는 GST효소활성을 측정 한 결과 정상군에 비해 1주간 매 일 300 mg/kg의 용량으로 시료 투여한 군이 66%정도 감소된다는 결과를 볼 수 있었다. glutathione 합성에 관여하는 $\gamma$-GCS의 활성과 산화형 glutathione을 환원형 glutathione으로 환원시키는 GR의 활성을 관찰한 결과 가자 추출물 투여군이 정상군보다 GR의 활성은 80% 감소되었고, 합성에 관여하는 $\gamma$-GCS의 활성은 정상군과 비교할 때 약간의 감소만을 나타내 glutathione함량 변동에 크게 영향을 미치지 않는 것으로 생각된다.

• 약학회지
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• 제40권3호
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• pp.326-334
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• 1996

Effects of methanol extract of Astragali Radix (AR) on the immune responses were studied using ICR mice. Mice were divided into 4 groups (10mice/group), and methanol extracts of AR at doses of 0.05, 0.25 and 1.25g/kg were orally administered to ICR mice once a day for 2 weeks. Mice were immunized and challenged with sheep red blood cells (SRBC). The results of this study were summarized as follows; (1) Methanol extract of AR at 0.05, 0.25 and 1.25g/kg didn't affect the weight ratios of thymus to body, as compared with those in controls, but significantly increased spleen weight ratio. (2)Methanol extract of AR at 0.05 and 0.25g/kg significantly increased hemagglutination titer and splenic plaque forming cells corresponding to humoral immunity, as compared with those in controls, but their enhancements were somewhat lowered at a high dose (1.25g/kg). (3) Methanol extract of AR at 0.05 and 0.25g/kg siginificantly increased delayed-type hypersensitivity reaction resulted from cell-mediated immunity, as compared with those in controls, but not so significant increases were observed at a high dose (1.25g/kg). (4) Methanol extract of AR at 0.05 and 0.25g/kg significantly increased phagocytic activity and the number of circulating leukocyte compared with those in controls, but their enhancements were lowered at a high dose (1.25g/kg). These results suggest that methanol extract of Astragali Radix increased humoral and cell-mediated immune responses, phagocytic activity and the number of circulating leukocyte, dependent upon dose, but inhibited their enhancement effects were decreased at a high dose (1.25g/kg).

• KSBB Journal
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• 제24권2호
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• pp.170-176
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• 2009

한국산 장류콩 종자로부터 최종적으로 Sephadex G-100상에서 분리한 lectin의 분자량, 적혈구 응집력, 온도, 열 안정성, 및 pH의 생화학적 특성을 조사하였다. 분리된 lectin을 SDS-PAGE한 결과, 분자량이 32 kDa와 22 kDa인 2개의 band가 나타났으며, Native PAGE에 의해서 108 kDa의 tetramer임을 알 수 있었다. Lectin은 trypsin을 처리한 사람의 A, B, AB, O 혈액과 쥐 혈액에서 혈구응집 반응이 일어나지 않았으나 토끼에서는 응집 반응이 일어났으며, trypsin을 처리하지 않은 모든 혈액에서는 응집반응이 일어나지 않았다. 이 lectin은 최적온도가 20-$50^{\circ}C$이며, 10-$60^{\circ}C$ 에서 열 안정성을 보였다. 최적 pH는 7.2로서 6.2 이하와 8.0 이상에서는 활성이 상설되었다.

• 약학회지
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• 제36권4호
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• pp.303-314
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• 1992

Experiments were performed on mice to investigate the effect of methionine diets (MET) on the immunotoxicity of ethanol. ICR female mice were divided into 5 groups, Met (Basal (B)+0.19% methionine(M), B+1.71% M and B+5.13%W) and ethanol(4%) were administered ad libitum for 21 days. The mice were evaluated for changes in immune status as measured by antibody titer, Arthus reaction, delayed type hypersensitivity (DTH), rosette forming cell(RFC) and plaque forming cell (PFC) to sheep red blood cells (S-RBC). To investigate the change of the non-specific immune response, the number of leukocytes in peripheral blood and phagocyte activity were measured. The results were summarized as follows: (1) The weight ratios of spleen and thymus to body weight were significantly increased by the B+0.19% M, B+0.57% M and B+1.71% M groups in comparison with control group(B), but B+5.13% M group was significantly decreased. (2) Humoral immune responses were significantly increased by the B+0.19% M and B+0.57% M groups in comparison with control group, but B+5.13% M group was significantly decreased. (3) Cellular immune responses were significantly decreased by the B+1.71% M and B+5.13% M groups in comparison with control group. (4) Phagocyte activities were significantly increased by the B+0.19% M, B+0.57% M and B+1.71% M groups in comparison with control groups, but B+5.13% M group was significantly decreased. (5) The number of circulating leukocyte was significantly increased in the B+0.19% M and B+0.57% M groups in comparison with control group, but B+5.13% M group was significantly decreased.

• The Korean Journal of Physiology and Pharmacology
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• 제17권6호
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• pp.547-551
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• 2013

We recently reported a Philyra pisum lectin (PPL) that exerts mitogenic effects on human lymphocytes, and its molecular characterization. The present study provides a more detailed characterization of PPL based on the results from a monosaccharide analysis indicating that PPL is a glycoprotein, and circular dichroism spectra revealing its estimated ${\alpha}$-helix, ${\beta}$-sheet, ${\beta}$-turn, and random coil contents to be 14.0%, 39.6%, 15.8%, and 30.6%, respectively. These contents are quite similar to those of deglycosylated PPL, indicating that glycans do not affect its intact structure. The binding properties to different pathogen-associated molecular patterns were investigated with hemagglutination inhibition assays using lipoteichoic acid from Gram-positive bacteria, lipopolysaccharide from Gram-negative bacteria, and both mannan and ${\beta}$-1,3-glucan from fungi. PPL binds to lipoteichoic acids and mannan, but not to lipopolysaccharides or ${\beta}$-1,3-glucan. PPL exerted no significant antiproliferative effects against human breast or bladder cancer cells. These results indicate that PPL is a glycoprotein with a lipoteichoic acid or mannan-binding specificity and which contains low and high proportions of ${\alpha}$-helix and ${\beta}$-structures, respectively. These properties are inherent to the innate immune system of P. pisum and indicate that PPL could be involved in signal transmission into Gram-positive bacteria or fungi.

• 동아시아식생활학회지
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• 제23권1호
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• pp.23-30
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• 2013

The purpose of this study was to evaluate the effects of several types of bibimbab (a Korean traditional meal of mixed rice with assorted vegetables), on various immune activities. Compared to control animals in a mouse model (given hamburgers), the oral administration of a portion of bibimbab containing wild plants significantly increased splenic B/T, thymic Th lymphocyte subpopulations, serum IFN-${\gamma}$ production, and enhanced hemagglutination titers up to 300%. Also, a consumption of mushroom-bulgogi bibimbab and Jeonju-style bibimbab markedly decreased compound 48/80-induced systemic anaphylaxis (immediate hypersensitivity), while bibimbab with wild plants inhibited SRBC-induced delayed type hypersensitivity. These results suggest that bibimbab with wild plants both up-regulate on immune activities and have anti-allergenic properties.

• Journal of Microbiology and Biotechnology
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• 제24권5호
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• pp.704-713
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• 2014

The highly pathogenic avian influenza A (HPAI) viruses of the H5N1 subtype infect poultry and have also been spreading to humans. Although new antiviral drugs and vaccinations can be effective, rapid detection would be more efficient to control the outbreak of infections. In this study, a phage-display library was applied to select antibody fragments for HPAI strain A/Hubei/1/2010. As a result, three clones were selected and sequenced. A hemagglutinin inhibition assay of the three scFvs revealed that none exhibited hemagglutination inhibition activity towards the H5N1 virus, yet they showed a higher binding affinity for several HPAI H5N1 strains compared with other influenza viruses. An ELISA confirmed that the HA protein was the target of the scFvs, and the results of a protein structure simulation showed that all the selected scFvs bound to the HA2 subunit of the HA protein. In conclusion, the three selected scFVs could be useful for developing a specific detection tool for the surveillance of HPAI epidemic strains.

• 대한수의학회지
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• 제61권2호
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• pp.13.1-13.8
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• 2021

Mammalian reovirus (MRV) causes respiratory and intestinal disease in mammals. Although MRV isolates have been reported to circulate in several animals, there are no reports on Korean MRV isolates from wildlife. We investigated the biological and molecular characteristics of Korean MRV isolates based on the nucleotide sequence of the segment 1 gene. In total, 144 swabs from wild animals were prepared for virus isolation. Based on virus isolation with specific cytopathic effects, indirect fluorescence assays, electron microscopy, and reverse transcription-polymerase chain reaction, only one isolate was confirmed to be MRV from a Korean roe deer (Capreolus pygargus). The isolate exhibited a hemagglutination activity level of 16 units with pig erythrocytes and had a maximum viral titer of 10^5.7 50% tissue culture infectious dose (TCID₅₀)/mL in Vero cells at 5 days after inoculation. The nucleotide and amino-acid sequences of the partial segment S1 of the MReo2045 isolate were determined and compared with those of other MRV strains. The MReo2045 isolate had nucleotide sequences similar to MRV-3 and was most similar (96.1%) to the T3/Bat/Germany/342/08 strain, which was isolated in Germany in 2008. The MReo2045 isolate will be useful as an antigen for sero-epidemiological studies and developing diagnostic tools.