• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
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• pp.261-261
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• 1994

Bovine milk xanthine oxidase (E.C.1.1.3.22, XO) purchased from Sigma Chemical Co. had the three protein fragments below 150 kDa on 7.5％ SDS-PAGE, which did not show enzyme activity. To remove these fragments, the enzyme preparation was further purified through Sephadex G-200 column chromatography. Two peaks exhibiting enzymatic activity were separated very closely to the void volume, which were revealed as two different enzyme forms, dimeric and monomeric, confirmed by activity staining on native PAGE. Anti sera-against each of the two enzyme forms were raised by subcutaneous injection at multiple sites on the back of rabbits during 4 weeks. On the immunodiffusion test, it was found that both of the antisera of the two forms could react with each other, which implied that their epitopes were identical In the Western blot analysis of mouse liver cytosol fraction, it was found that rabbit anti-XO antibody bound well with the protein band of monomeric mouse liver XO of about 150kDa. Based on this result, mouse liver cDNA 1 ibrary was screened by in situ hybridizat ion wi th rabbi t anti -XO antibody as probe. Through the immunological screening, recombinant phages giving positive signal by the production of XO were selected and further purified. To validate these clones, purified phages were lysogenized in E. coli Y1089 and their lysates were analysed for enzyme activity and immunoreactivity, It was verified that lysates of the purified recombinant phage lysogens exhibited the enzymatic activity as well as bound wi th XO antibody, when induced by IPTG. The above results assert that selected recombinant phage carries mouse liver XO gene.

• Journal of Biomedical and Translational Research
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• 제19권4호
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• pp.92-102
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• 2018

The objectives of this study were to find out the plant to enhance immune activity among 42 kinds of foods frequently consumed by the Korean elderly consisting of 5 food groups and 5 wild plants. Each sample was assessed the immunoactive effect by measuring $NF-{\kappa}B/AP1$ gene expression, nitric oxide and cytokine production in $RAW-Blue^{TM}$ cell. Soybean sprouts of 47 plants showed the highest $NF-{\kappa}B/AP1$ gene expression at the level of $1.13{\pm}0.03$ (O.D. 650 nm) and Soritae, sweet potato, banana, apple, garlic, crown daisy, cabbage and Ailanthus altissima also had high activity of $NF-{\kappa}B/AP1$ gene in $RAW-Blue^{TM}$ cell stimulated by LPS. NO production of Ailanthus altissima was significantly higher than that of other plants and 16 plants of glutinous sorghum, black rice, Seoritae, Heuktae, sweet potato, banana, apple, garlic, mungbean sprouts, spinach, crown daisy, young pumpkin, cabbage, soybean sprouts, Actinidia arguta and Aster scaber were the next best activity. The above results selected 17 out of 47 plant samples. Moreover, soybean sprouts was significantly shown to increase $TNF-{\alpha}$ ($1,509.55{\pm}1.38pg/mL$) and $IL-1{\beta}$ ($54.56{\pm}1.08pg/mL$) cytokines in comparison with RAW-Blue cell stimulated by LPS. According to the results of in vitro evaluation, the ethanol extract of soybean sprout increased the production of immune-enhancing cytokines by proliferation of macrophages. In addition, $NF-{\kappa}B$ transcription factor activity and NO production ability were excellent, and it was selected as a material having excellent immunological activity.

• 한국패류학회지
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• 제31권3호
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• pp.187-194
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• 2015

본 연구에서는 수온 증가에 따른 말전복 (Haliotis gigantea) 과 둥근전복 (H. discus discus) 헤모림프의 생리 및 면역학적 변화를 관찰하기 위하여 위 두 전복을 20, 22, 24, 26 및 $28^{\circ}C$ 수온에 각각 4 일간 노출시켰다. 노출 결과, 헤모림프의 totalo-protein (TP), glucose, calcium (Ca) 은 둥근전복 (H. discus discus)이 말전복 (H. gigantea) 보다 높은 값을 보였으나, magnesium (Mg), alkaline phosphatase (ALP) 두 전복에서 유사한 값을 보였다. 그리고 면역인자인 PO에서는 둥근전복 (H. discus discus)이 더 높게 나타났다. 수온 증가에 따른 헤모림프의 TP, glucose, Mg, AST, ALT 및 ALP는 유의적인 변화가 나타나지 않았다. 하지만 헤모림프의 Ca 농도는 둥근전복 (H. discus discus)에서 $24^{\circ}C$ 이상, 말전복 (H. gigantea)에서는 $26^{\circ}C$ 이상의 수온에서 유의적 증가를 보였다. 그리고 면역인자인 phenoloxidase (PO) 는 둥근전복 (H. discus discus)은 $22^{\circ}C$ 이상 말전복 (H. gigantea) 에서는 $26^{\circ}C$ 이상의 수온에서 유의적 감소를 나타내었다. 결론적으로 말전복 (H. gigantea)은 $24^{\circ}C$ 이상에서 혈액학적 변화를 보였고, $26^{\circ}C$ 이상에서 면역학적 변화를 보였다. 둥근전복 (H. discus discus)은 $26^{\circ}C$ 이상에서 혈액학적 변화를 보였고, $22^{\circ}C$ 이상에서 면역학적 변화를 나타내었다.

• 한국식품과학회지
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• 제34권2호
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• pp.325-329
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• 2002

남해안에서 다량 생산되고 있는 김(Porphyra yezoensis)의 생리활성 물질은 porphyran을 분리 추출한 후, 흰쥐에서 고지혈증 및 고 콜레스테롤혈증을 유발시켜 간장의 항산화 효소활성 및 면역능을 측정하였다. 간의 효소활성 측정을 위해 Sprague-Dawley계 흰쥐를 9마리씩 5군으로 나누어 1군은 정상군, 2군은 대조군(고지혈증 유발군), 3, 4 및 5군은 porphyran 급이군으로서 대조군에 porphyran을 각각 1%, 5% 및 10%의 비율로 사료에 첨가하여 4주간 사육하였다. 면역능은 Balb/c female mouse를 이용하여 3마리씩 4군으로 대조군 및 porphyran 1%, 2% 및 5%로 조제된 시료를 실험동물에게 2일에 1회 1mL씩 복강 주사를 연속적으로 20일간 투여한 후 비교하였다. 혈청중의 alanine aminotransferase(ALT), aspartate aminotransferase(AST), alkaline phosphatase(ALP) 및 lactic dehydrogenase(LDH) 활성은 대조군에 비해 porphyran 급이군에서 낮았으며, porphyran의 급이 수준이 증가할수록 대조군에 비해 유의하게 낮은 경향을 보였다. 간장의 superoxide dismutase와 catalase활성은 정상군에 비하여 대조군이 유의하게 높았으나, 대조군에 비해 porphyran 급이군에서는 낮았으며, 특히 10% porphyran 급이군에서는 정상군 수준으로 유의하게 감소되었다. TBARS 역시 대조군에 비해 porphyran 급이군 모두 유의하게 감소하였으며, 정상군 수준이하로 감소되었다. Interleukin-1농도는 porphyran 2% 및 5%를 투여한 흰쥐의 혈청에서 정상군보다 각각 26.81%, 77.95%의 증가를 나타내었으나, Interleukin-2는 감소하였다. 본 연구 결과에서 porphyran 급이는 고지혈증을 유발한 흰쥐에서 과산화물 농도를 저하시켜 항산화 효소활성을 낮추었으며, 면역능에서 일정한 현상은 나타나지 않았다.

• 한국패류학회지
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• 제31권1호
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• pp.1-8
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• 2015

본 연구에서는 수온 증가에 따른 북방전복 (Haliotis discus hannai) 과 둥근전복 (H. discus discus) hemolymph의 생리 및 면역학적 변화를 관찰하기 위하여 위 두 종을 20, 22, 24, 26 및 $28^{\circ}C$ 수온에 각각 4 일간 노출시켰다. 노출 결과, hemolymph의 total-protein (TP), glucose, 칼슘 (Ca) 은 둥근전복이 북방전복보다 높은 값을 보였으나, 마그네슘 (Mg), alkaline phosphatase (ALP) 및 lysozyme은 두 종에서 유사한 값을 보였다. 수온 증가에 따른 hemolymph의 TP, glucose 및 마그네슘은 두 전복 모두에서 유의한 변화가 관찰되지 않았다. 그러나 칼슘과 면역관련 인자인 ALP 및 lysozyme은 둥근전복에서 유의한 차이가 없었으나, 북방전복에서는 $26^{\circ}C$ 또는 $28^{\circ}C$의 고수온에 노출되었을 때, 유의하게 높은 활성을 보였다. 한편 phenoloxidase (PO)는 북방전복에서 높은 값을 보였으며, 수온이 높을수록 증가하는 경향을 보였다. 결론적으로 북방전복은 둥근전복과 달리 여러 지표에서 유의한 변화가 관찰된 것으로 보아 북방전복이 둥근전복보다 고수온에 보다 민감한 것으로 사료된다.

• 한국균학회지
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• 제39권3호
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• pp.205-212
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• 2011

진균의 phenylalanine ammonia-lyase(PAL)와 식물의 PAL간 그리고 PAL과 histidine ammonia-lyase (HAL)간의 구조적 특성 관계에 대한 이해를 얻기 위해 옥수수병원균인 깜부기균(Ustilago maydis)의 PAL에 대한 면역학적 분석을 수행하였다. 진균의 PAL 항체와 식물의 PAL 항체를 이용하여 분석하였을 때 실험한 모든 종의 PAL 간에 교차반응이 나타났다. 알팔파 PAL 항체와 포플라 PAL 항체 모두 식물 PAL은 강하게 인식하였으나 진균의 PAL은 약하게 인식하였다. U. maydis PAL은 Rhodotorula glutinis 효모의 PAL만 약하게 인식하였다. U. maydis PAL은 식물의 PAL에 대해서 낮은 친화성을 보였으나 Pseudomonas 세균의 HAL에 대해서는 강한친화성을 보였다. 2종의 식물 PAL 항체들 또한 Pseudomonas 세균의 HAL에 대해 강한 친화성을 보였다. 진균과 식물의 PAL 항체는 PAL 효소의 활성 저해를 나타냈으며 세균의 HAL 효소의 활성에 대해서도 중도적 저해를 나타냈다. 세균의 HAL 항체는 식물과 효모와 Ustilago PAL 중에 Ustilago PAL 활성만 저해하였다. 식물의 PAL과 진균의 PAL 모두 효모의 PAL 활성은 저해하지 못했다. 본 연구는 PAL과 HAL간에 면역학적 관계가 있음을 처음으로 보고한다.

• The Korean Journal of Physiology and Pharmacology
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• 제14권2호
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• pp.71-75
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• 2010

To investigate the intestinal absorption of a fibrinolytic and proteolytic lumbrokinase extracted from Eisenia andrei, we used rat everted gut sacs and an in situ closed-loop recirculation method. We extracted lumbrokinase from Eisenia andrei, and then raised polyclonal antibody against lumbrokinase. Fibrinolytic activity and proteolytic activity in the serosal side of rat everted gut sacs incubated with lumbrokinase showed dose- and time-dependent patterns. Immunological results obtained by western blotting serosal side solution using rat everted gut sacs method showed that lumbrokinase proteins between 33.6 and 54.7 kDa are absorbed mostly by the intestinal epithelium. Furthermore, MALDI- TOF mass spectrometric analysis of plasma fractions obtained by in situ recirculation method confirmed that lumbrokinase F1 is absorbed into blood. These results support the notion that lumbrokinase can be absorbed from mucosal lumen into blood by oral administration.

• 대한수의학회지
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• 제54권4호
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• pp.203-208
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• 2014

Bone marrow is a hematological and immunological organ that provides multiple immune cells, including B lymphocytes, and thus plays a critical role in the efficacy of vaccine. We previously demonstrated that Bordetella (B.) bronchiseptica antigen has high immunogenicity in spleen cells, a peripheral immune organ. In this study, we investigated the immunogenicity of B. bronchiseptica antigen in bone marrow cells, a central immune organ. B. bronchiseptica antigen increased the cellular activity of bone marrow cells and significantly enhanced the production of nitric oxide, IL-6, and TNF-${\alpha}$. Bone marrow cells primed with B. bronchiseptica antigen in vivo were harvested and stimulated with the same antigen in vitro. The stimulation of B. bronchiseptica antigen significantly increased the cellular activity and proliferation rate of the primed cells. B. bronchiseptica antigen also greatly induced the production of antigen-specific antibody in the primed cells. Taken together, the present study demonstrated that B. bronchiseptica antigen can stimulate bone marrow cells, a central immune organ, and recall the immune response of the primed bone marrow cells.

• Asian-Australasian Journal of Animal Sciences
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• 제20권9호
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• pp.1438-1443
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• 2007

The experiment was conducted to evaluate the effects of Ligustrum lucidum (LL) and Schisandra chinensis (SC) on the growth, antioxidative metabolism and immunity of laying strain male chicks. The results showed that diets supplemented with 1% of either LL or SC had no effects on the growth performance of chicks compared with the control. Furthermore, both LL and SC significantly reduced malondialdehyde (MDA) concentration of serum and heart of chicks (p<0.05). In addition, superoxide dismutase (SOD) activity of serum of the birds was significantly elevated by supplementation with SC (p<0.05). Glutathione reductase (GR) activity of heart and serum of the birds was significantly elevated by supplementation with LL or SC (p<0.05). LL supplementation significantly elevated antibody values against Newcastle Disease virus (NDV)(p<0.05) and lymphoblastogenesis (p<0.05) of the birds. The results suggest that diets supplemented with 1% of either LL or SC may improve immune function and antioxidant status of chicks.

• BMB Reports
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• 제39권6호
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• pp.671-676
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• 2006

The holoenzyme of protein phosphatase (PP) from tulip petals was purified by using hydrophobic interaction, anion exchange and microcystin affinity chromatography to analyze activity towards p-nitrophenyl phosphate (p-NPP). The catalytic subunit of PP was released from its endogenous regulatory subunits by ethanol precipitation and further purified. Both preparations were characterized by immunological and biochemical approaches to be PP2A. On SDS-PAGE, the final purified holoenzyme preparation showed three protein bands estimated at 38, 65, and 75 kDa while the free catalytic subunit preparation showed only the 38 kDa protein. In both preparations, the 38 kDa protein was identified immunologically as the catalytic subunit of PP2A by using a monoclonal antibody against the PP2A catalytic subunit. The final 623- and 748-fold purified holoenzyme and the free catalytic preparations, respectively, exhibited high sensitivity to inhibition by 1 nM okadaic acid when activity was measured with p-NPP. The holoenzyme displayed higher stimulation in the presence of ammonium sulfate than the free catalytic subunit did by protamine, thereby suggesting different enzymatic behaviors.