• Korean Journal of Food Science and Technology
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• v.36 no.6
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• pp.986-990
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• 2004

The immunomodulatory effect of Codonopsis lanceolata based on the production of cytokines and the activation of macrophage was studied. The mRNA expression of nitric oxide synthase (iNOS) was gradually induced after 24 hr treatment of Codonopsis lanceolata, and NO production was a maximum after 24 hr treatment with 1 mg/mL. RAW 264.7 cell on in vitro treatment with Codonopsis lanceolata induced mRNA of cytokines such as interleukin-1(IL-1)${\beta}$, interleukin-6(IL-6), tumor necrosis $factor(TNF)-{\alpha}\;and\;interferon(IFN)-{\gamma}$; $IL-1{\beta}$ and IL-6 mRNA were gradually induced up to 24 hr, $TNF-{\alpha}\;mRNA$ was regularly induced up to 24 hr, and $IFN-{\gamma}\;mRNA$ level was a maximum within 1 hr. These results suggest that Codonopsis lanceolata exerts as an effective immunomodulator and enhances antitumor activity of macrophages.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.5
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• pp.664-672
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• 2015

In order to develop new immuno-stimulating ingredients from mature leaves of green tea, crude polysaccharides were isolated from pectinase digests of tea leaves (green tea enzyme digestion, GTE-0), after which their immuno-stimulating activities and chemical properties were examined. GTE-0 mainly contained neutral sugars (54.9%) such as glucose (14.2%), arabinose (12.2%), rhamnose (11.1%), and galacturonic acid (45.1%), which are characteristic of pectic polysaccharides. The anti-complementary activity of GTE-0 was similar to that of polysaccharide K (used as positive control). Number of morphologically activated macrophages was significantly increased in the GTE-0-treated group. GTE-0 significantly augmented $H_2O_2$ and reactive oxygen species production by murine peritoneal macrophage cells in a dose-dependent manner, whereas production of nitric oxide showed the highest activity at a dose of $100{\mu}g/mL$ among all tested concentrations. Murine peritoneal macrophages stimulated with GTE-0 showed enhanced production of various cytokines such as interleukin (IL)-6, IL-12, and tumor necrosis factors-${\alpha}$ in a dose-dependent manner. Further, GTE-0 induced higher phagocytic activity in a dose-dependent manner. In ex vivo assay for cytolytic activity of murine peritoneal macrophages, GTE-0-treated group showed significantly higher activity compared to the untreated group at an effector-to-target cell ratio of 20. The above results lead us to conclude that polysaccharides from leaves of green tea have a potent immuno-stimulating effect on murine peritoneal macrophage cells.

• Proceedings of the Korean Society of Life Science Conference
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• 2002.05a
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• pp.17-22
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• 2002

지구상에는 수천종의 버섯류가 자생하고 있어 유전자원으로서의 중요성이 지대할 뿐만 아니라 기능성 식품소재 및 각종 약리 활성을 나타내는 신약개발 소재로도 크게 주목을 받고 있다. 이들 버섯은 균사체의 영양대사로 얻어지는 대사산물이 축적된 자실체의 형태로 나타나는데, 최근에 와서 자실체 및 균사체의 추출물이나 균사체 배양물이 체질개선이나 각종 병의 예방과 치료에 효과가 있는 것으로 밝혀져 건강식품이나 의약품으로서의 용도가 크게 증가하고 있는 실정이다. 특히, 담자균이 생산하는 특정 구조를 갖는 다당류는 오래전부터 종래의 화학요법제와는 달리 숙주내의 면역 기능을 부활하여 소위 면역요법제로서의 항암효과를 나타냄이 알려져왔었다. 현재까지 제약 및 의학적인 방법이 질병의 주된 치료방법으로 이용되어 왔지만 최근에 특정식품의 섭취가 만성질환의 발생을 억제 또는 지연시킨다는 연구 보고가 나오면서부터 만성질환의 치료방법으로서 식이요법을 중요하게 생각하게 되었다. 따라서 새로운 식품소재 및 가공식품의 개발을 통한 성인병 등의 각종 질병예방이 국민보건문제 해결에 필수적이다. 현재 일본 등에서는 표고버섯, 구름버섯 및 치마버섯 유래의 다당체 또는 단백다당체인 lentinan, krestin 또는 PS-K, schizophyllan 및 PSP 등이 실용화되어 높은 가격에 판매되고 있다. 국내에서도 야생 구름버섯 자실체로부터 추출한 단백 다당체인 Copolang(광동제약)이 개발되어 PS-K와 유사하게 암의 치료에 병행 사용되고 있고, 또 강력한 항암활성이 보고된 상황버섯의 균사체 추출물인 단백 다당체가 Mesima-Ex FK(한국신약)라는 상품명으로 암의 치료에 병행 사용되고 있는 것으로 알려지고 있다. 담자균류와 아울러 미생물 유래 다당체는 그 구조와 특성에 있어서 매우 다양함을 지니고 있다. 이러한 미생물 유래 다당류의 공업적 생산과 이용에 대한 연구로서는 Leuconostoc mesenteroides가 생산하는 dextran이 혈장증량제로 개발된 이래 Xanthomonas campestris가 생산하는 pullulan, Zoogloea rgmigera가 생산하는zooglan둥이 대표적인 예로 보고되고 있다. 한편, 미생물 유래 다당류는 구성당, 분자량, 화학적 구조 등과 같은 특성의 차이에 의해 많은 종류가 존재하고 있으며, 다양한 물성 및 유화제, 응고제, gel 형성제, 필름 형성제, 흡착제, 안정제, 접착제 등과 같은 용도로 광범위하게 이용되고 있다. 또한 근래에 들어서는 미생물 유래 다당체가 지니는 항암활성이 확인되어 새로운 의약품으로서의 개발 가능성이 기대된다. 그 밖에도 기존에 알려져 있는 식물 및 해조류 유래의 다당체와는 달리, 발효조를 이용한 연속배양에 의해 공업적 대량 생산이 가능하며, 더욱이 생산된 다당체의 분리 및 회수가 용이하다는 이점을 지니고 있다. 최근에 들어서는 유전공학적 기법을이용한 고생산성 변이균주 및 새로운 기능을 지닌 다당체의 개발에 관한 연구가 보고되고 있는 등 고부가가치를 지닌 새로운 바이오 소재로서의 기능 및 용도 개발에 관한 연구가 활발히 진행되고 있다. 이러한 항암 활성을 나타내는 여러 가지 담자균류중 Agaricus blazei로부터 생산되는 다당체는 고형암 이외에 S형 결장암, 난소암, 유방암, 폐암, 간암 등에 효과가 입증되었고, 천연물질에 의한 암 면역요법으로 각광을 받고 있으며, 항암 및 항virus의 완치율과 저지율에서 현재 여러가지 약효가 있는 버섯중에서도 탁원한 효과가 있는 것으로 증명되고 있다. 이들 다당체는 사이토카인을 생산시켜서 T임파구와 B임파구의 항원 특이적인 면역반응을 활성화시키고, 세포장해성 T세포와 활성화 대식세포의 세포장해 기능을 충진시켜서 암세포를 파괴시킨다. 또한 콜로니 자극인자인 사이토카인을 생산시켜서 면역담당세포의 신생을 촉진시키기도 하며, 암의 화학요법과 방사선 요법으로 저하된 백혈구를 회복시키는 역할을 한다. 따라서 최근의 연구동향은 생산된 다당체의 항암활성을 향상시키고자 하여 배양기간중에 interleukin을 의도적으로 첨가하는 경향이 있다. 이러한 항암 활성을 나타내는 담자균체 유래 다당체는 버섯의 기원에 따라 그 형태에 약간의 차이를 나타내기는 하나 그 기본 형태는 ${\beta}-(1,6)-glucosyl$ 분지를 가진 ${\beta}-(1,3)-glucan$이며, 평균 분자량은 50 ${\sim}$ 200만 정도이다. Agaricus blazei의 원산지인 브라질의 피에다데(Piedade) 지방의 환경조건(산지의 습도는 80%, 낮 기온 $35^{\circ)C$, 밤 기온 $20{\sim}25^{\circ}C$로 대단히 높으며, 정기적으로 열대지방 특유의 소나기가 내리는 지역)에서 볼 수 있듯이 Agaricus blazei의 성장 환경은 매우 까다로운 편이며, 날것으로는 보관이 잘 안되기 때문에 그 재배에 큰 어려움이 있다. 또한, 고체배양에 의해 생산된 버섯 자실체로부터 유기용매 및 열수추출 방법으로 다당체를 생산하는 방법은 균일한 형태의 버섯 자실체를 공급받기가 어렵기 때문에 다당체의 생산 수율이 낮고, 많은 노동력이 요구되는 어려움이 있다. 그러나 액체배양에 의한 다당체 생산의 경우는 고체배양에 의한 다당체 생산에 비해 일정한 조건하에서 배양이 가능하다는 장점이 있으며, 항상 균일한 균사체 및 배양액을 얻을 수 있다. 따라서 원하는 유용물질을 쉽게 획득할 수 있는 장점이 있다.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.12
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• pp.1930-1939
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• 2013

It has been reported that Chungkukjang, one of Korean traditional fermented soybean products, may improve hypertension, diabetes, and hyperlipidemia. In this study, we sought to investigate the immunoenhancing effects of Chungkukjang in ovariectomized mice. For the first period, female SLC ddy mice were either sham-operated (Sham; n=27) or ovariectomized (OVX; n=27). As a basal diet, ovariectomized mice were fed low-calcium diet for faster induction of osteoporosis for six weeks, and those in the Sham group were fed AIN-76 diet. For the second period, half of the OVX group (n=9) and the Sham group (n=9) were fed a Chungkukjang-based diet (CKJ); whereas the other half (OVX; n=9/ Sham; n=9) were fed a casein-based diet (CSI) for 8 weeks. After a second period, we collected the blood via heart puncture and measured the splenocytes proliferation, T lymphocyte subsets by flowcytometry, and levels of serum cytokines (IL-2, IL-4, IL-6, IL-10, IFN-${\gamma}$ and TNF-${\alpha}$) by ELISA assay. The OVX+CKJ group showed higher splenocytes proliferation, higher ratio of CD4/CD8, and lower levels of IL-6 and TNF-${\alpha}$ cytokines compared to the OVX+CSI group. The Sham+CKJ group showed cytokine productions, such as higher levels of IL-10 and IFN-${\gamma}$, and lower levels of IL-6 and TNF-${\alpha}$ compared to the Sham+CSI group. The result of this study suggests that Chungkukjang may lower the proinflammatory cytokine levels in both the OVX and Sham groups. In addition, Chungkukjang could make a balance of T cell subset proliferations and enhance the splenocyte proliferations in the OVX group.

• Journal of Animal Science and Technology
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• v.49 no.5
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• pp.599-610
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• 2007

Effect of dietary krill meal levels on the cellular immunity was studied in broiler chicks activated immune response. One day old male broiler chicks(Ross) were fed the experimental krill meal 0.0(basal), 0.5, 1.0 and 2.0% diets for 3wks. Blood TNF-α activity, ovotransferrin level and Con A induced proliferation of PBMC and splenocytes after 24 hr(21 d age) of the croton oil 10㎕ injection intra- musculary at the age of 20 days compared to the control olive oil. Krill meal diets did not affect growth performance of broiler chicks and plasma ovotransferrin levels but decreased significantly(p<0.0001) TNF-α like activity and proliferation of PBMC relative to krill meal 0.0% diet. And the proliferation of splenocytes were significantly(p<0.05) increased in birds fed krill meal 1.0% diet relative to krill meal 0.5 and 2.0% diets. The croton oil injection induced a significant(p<0.0001) increases in the TNF-α activity or the PBMC proliferation and enhanced circulating ovotransferrin levels relative to the olive oil. In birds injected with the croton oil the proliferation of PBMC was reduced linearly with the increase of dietary krill meal levels, and the proliferation of splenocytes was decreased in the krill meal 1.0 and 2.0% diets relative to olive oil. These results indicated that dietary krill meal changed the innate and cellular immunity in broiler chicks activated by the injection of croton oil.

• The Journal of the Korean bone and joint tumor society
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• v.5 no.3
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• pp.162-168
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• 1999

The products of c-fos and c-jun proto-oncogenes form the heterodimeric complex activator protein 1 (AP-1), which plays an important part in the control of bone cell proliferation and differentiation, as well as in the development of bone tumors. The expression of c-fos protein was examined in 35 cases of human osteosarcomas as formalin-fixed paraffin-embedded tissue sections using a monoclonal antibody. The expression of c-fos was restricted to bone-forming lesions, while low grade cartilaginous tumors were devoid of immunoreactivity. The highest levels of c-fos expression were detected in osteoblastic osteosarcoma (13 of 17 cases with grade one on two) while two chondroblastic osteosarcomas, one fibroblastic osteosarcoma, and two parosteal osteosarcomas were negative. Two cases of telangiectatic osteosarcomas were positive for c-fos protein. However, since there is a tendency of high c-fos protein expression at the higher histological grade, significant differences were not present in the expression of c-fos protein. Thus c-fos expression may be implicated in the development of osteosarcomas, but they appear to have little or no relevance in the development of low grade cartilaginous neoplasms.

• KSBB Journal
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• v.14 no.1
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• pp.82-90
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• 1999

In order to eventually fabricate an analytical system for infectious microorganisms, we synthesized major immunochemical components, utilized them for the construction of model system, and investigated an assay concept for bacterial whole cells. For the preparation of system components, a polyclonal antibody, against Salmonella thompson as model analyte, purified by immuno-affinity chromatography was used to chemically link to streptavidin or an enzyme, horseradish peroxidase(HRP). The antibody and streptavidin was modified with sulfosuccinimidyl 4-[N-maleimidomethyl]cyclohexane-1-carboxylate and N-succinimidyl-3-[2-pyridyldithio]propionate(subsequently activated by dithiotheritol), respectively. The modified components were reacted to synthesize antibody-streptavidin conjugates which were then purified on a two-layer chromatography column of diaminobiotin gel and Sephadex G-100. For antibody-HRP conjugates, HRP molecules were activated by $NalO_4$ oxidation and then coupled to immunoglobulin. After stabilizing with ($NaCNBH_3$, the conjugates were purified by size exclusion chromatography on Biogel A5M column. To devise a model system, such produced components were combined with a dot-blotter in which a nitrocellulose membrane($12{\mu}m$ pre size) with immobilized biotin was already located. The analyte (S. thompson cells) was reacted with the both antibody conjugates in a liquid phase, and the complexes formed were captured on the membrane surfaces by applying vacuum in the bottom compartment of the blotter to invoke biotin-streptavidin reaction. Under optimal conditions, the system enabled to identify the analytical concept for bacterial whole cells, and the lower limit of detection was approximately $1{\mu}g/m{\ell}$($10^5-10^6$ cells/m$m{\ell}$). The controlling factors were the concentrations of each antibody conjugate that caused agglutination in the presence of analyte as they increased.

• Journal of Life Science
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• v.22 no.6
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• pp.828-836
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• 2012

The objective of the current study was to determine the effects of the extracts isolated from the fruit body of Flammulina velutipes cultivated with oriental herbal plants on mouse splenocytes, B cells, and macrophages in vitro. The ethanol extracts B (EEB) directly induced the proliferation of spleen cells in a dose-dependent manner and increased IL-6, TNF-${\alpha}$, and IFN-${\gamma}$ synthesis. The EEB also increased the proliferation of B cells in a dose-dependent manner. The production of immunoglobulin M, G1, G2a, G2b, and IgG3 in the presence of the EEB increased progressively in the culture supernatant. When the EEB were used in macrophage cell line (RAW264.7) stimulation, there was a marked induction of NO synthesis in a dose-dependent manner and an increased IL-6, TNF-${\alpha}$, and GM-CSF synthesis. Intraperitoneal injection with EBB showed life prolongation effect of 16.1% in mice previously inoculated with sarcoma-180, respectively. These results suggest that the capacity of the EEB isolated from the fruit body of Flammulina velutipes cultivated with oriental herbal plants seems to act as a potent immunomodulator causing augmentation of immune cell activity, and with the absence of notable side-effects, Flammulina velutipes EEB could be used as a biological response modifier having possible therapeutic effects against immunological disorders. This study also showed that functional components of Flammulina velutipes were possibly improved by incorporating oriental herbal plants in a growth medium.

• Korean Journal of Food Science and Technology
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• v.39 no.3
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• pp.315-322
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• 2007

This study was carried out to investigate the functional activities of microorganisms isolated from kochujang, such as fibrinolytic, immunostimulating, and cytotoxical activities, and to apply these microorganisms to kochujang products. Ninety-one microbial strains with proteolytic activity were selected from 294 strains isolated from traditional and commercial kochujang. Three strains (TPP 0014, TPP 6013, and TPP 6015) with high fibrinolytic activity were tested for their immunostimulating and cytotoxical activities. For the assessment of macrophage activation, cytokines such as tumor necrosis factor, $interleukin-1{\alpha}$ and nitrogen oxide were measured with the murine macrophage cell line RAW 264.7. In addition, the cytotoxical activities of the three strains were examined by MTT assay on the colon cancer cell line SNU-C4 and normal cell line CHO-K1. Using an API identifying kit, two of the microbial strains (TPP 0014 and TPP 6015) were identified as Bacillus stearothermophilus and the other strain (TPP 6013) was identified as B. amyloliquefacience.

• Restorative Dentistry and Endodontics
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• v.32 no.6
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• pp.514-521
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• 2007

The purinoreceptor, $P2X_3$ is a ligand-gated cation channel activated by extracellular ATP. It has been reported that ATP can be released during inflammation and tissue damage, which in turn may activate $P2X_3$ receptors to initiate nociceptive signals. However, little is known about the contribution of $P2X_3$ to the dental pain during pulpal inflammation. Therefore, the purpose of this study was to investigate the expression of $P2X_3$ and its colocalization with TRPV1 to understand the mechanism of pain transmission through $P2X_3$ in the human dental pulp with double labeling immunofluorescence method. In the human dental pulp, intense $P2X_3$ immunoreactiyity was observed throughout the coronal and radicular pulp. Of all $P2X_3$-positive fibers examined, 79.4% coexpressed TRPV1. This result suggests that $P2X_3$ along with TRPV1 may be involved in the transmission of pain and potentiation of noxious stimuli during pulpal inflammation.