• 제목/요약/키워드: Hemolytic plaque assay

검색결과 10건 처리시간 0.022초

An antitumor component of laetiporus sulphureus and its immunostimulating activity

  • Kang, Chang-Yuil;Lee, Chong-Ock;Chung, Kyeong-Soo;Choi, Eung-Chil;Kim, Byong-Kak
    • Archives of Pharmacal Research
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    • 제5권2호
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    • pp.39-43
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    • 1982
  • A protein-polysaccharide fraction was prepared from the carpophores of Laetiporus sulphureus. This fraction suppressed growth of sarcoma 180 in A-strain mice when administered i. p. To investigate the mechanism of antitumor action of this fraction, plaque assay was conducted by administrating i. p. to the mise at a dose level of 50mg/kg for five days. Ten days later, the mice were immunized with 1 * 10$^{7}$ sheep red blooc cells. The number of hemolytic plaque forming cells was significantly greater than that of the control mice. Three monosaccharides and fifteen amino acids were identified in the protein-polysaccharide fraction.

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Effect of Kp, an Antitumor Protein-Polysaccharide from Mycelial Culture of Phellinus Linteus on the Humoral lammune Response of Tumor-Bearing ICR Mice to Sheep Red Blood Cells

  • Chung, Kyeong-Soo;Kim, Shin-Sook;Kim, Hee-Soo;Kim, Kye-Yang;Han, Man-Woo;Kim, Kil-Hyun
    • Archives of Pharmacal Research
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    • 제16권4호
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    • pp.336-338
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    • 1993
  • The immunomodulating activity of Kp, an antitumor protein-polysacchanide preparation from the shake-cultured mycelia of Phellinus linteus, was investigated in ICR mice subcutaneously implanted wit $1\times10^6$ cells of sarcoma 180. The mice were intraperitoneally administered with Kp at a does of 100 mg/kg once daily for five consecutive days starting from 24 hrs after the tumor implantation. Ten days after the last injection, the mice were immunized with $1\times10^7$ or $4\times10^8$ sheep red blood cells (SRBC) and five days later, the antibody-forming immune response were assessed by direct hemolytic plaque assay. To an immunization does of $1\times10^7$ SRBC, the Kp-treated mice elicied a successful humoral immune response despite the turmor-burden and produced $259\times10^3$ plaque-forming cells (PFC)/spleen, while the corresponding tumor-bearing control mice showed virtually no reponse $(2.0\times10^3$ PFC/spleen) (the stimulation index=129.5). However, to an immunization dose of $4\times10^8$ SRBC, both of the control mice and Kp-treated mice showed almost the same level of strong humoral immune response. From these data it is clear that Kp effectively restores the humoral immune response of the turmor-bearing ICR mice.

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한국산 고등 균류의 성분 연구(제73보) -좀우단버섯 배양 균사체의 항암 성분- (Studies on Constituents of Higher Fungi of Korea (LXXIII) Antitumor Components of the Cultured Mycelia of Paxillus atrotomentosus)

  • 곽상덕;복진우;현진원;최응칠;김병각
    • 한국균학회지
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    • 제20권3호
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    • pp.240-251
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    • 1992
  • 담자균류인 좀우단버섯 Paxillus atrotomentosus의 항암성분을 구명하기 위하여 그 균사체를 열수 추출하여 단백 다당체를 분리하였고, 이를 DEAE-cellulose ion 교환수지와 Sepharose CL-4B gel filtration chromatography로 정제하였다. 분획들을 각각 20 또는 50 mg/kg/day의 용량으로 마우스의 복강내에 투여하였을 때, sarcoma 180 고형암에 대하여 Fr. A가 68.51%의 가장 높은 억제율을 나타내었다. 항암기전을 밝히기 위하여 마우스에 대한 면역학적 실험을 한 결과, Fr. A는 대조군에 비해 활성화된 macrophage에서 분비되는 superoxide anion 양을 1.1배, hemolytic plaque assay에서의 용혈반 형성 세포수를 2.3배 증가시켰고 감소된 DTH 반응을 정상수준으로 회복시켰다. 암세포에 대한 직접적인 영향을 조사하기 위하여 MTT assay를 한 결과 대조군에 비해 세포의 viability가 약 32% 감소됨을 나타내었다. 화학분석에 의해, 이 성분은 fucose, galactose, glucose, mannose 및 xylose로 구성된 86.36%의 다당체와 14종의 아미노산으로 구성된 1.52%의 단백질 및 1.64% hexosamine으로 구성된 protein-bound polysaccharide이었다. 이 성분을 paxillan으로 명명한다.

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한국산 고등 균류의 성분 연구(제67보) -영지버섯 자실체의 항암성분- (Studies on Constituents of Higher Fungi of Korea (LXVII) -Antitumor Components of the Basidiocarp of Ganoderma lucidum-)

  • 현진원;최응칠;김병각
    • 한국균학회지
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    • 18권2호
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    • pp.58-69
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    • 1990
  • 담자균류인 영지 Ganoderma lucidum의 자실체를 열수추출한 후 얻은 단백 다당체를 DEAE cellulose 이온교환수지와 Sepharose CL-4B 젤 여과법을 이용하여 분리 정제하였다. 정제한 분획들을 각각 CR, IN, IA, GL과 GH로 명명하였다. 각각의 성분을 20mg/kg/day 용량으로 투여하였을 때 sarcoma 180 고형암에 대하여 Fr. GL이 81%의 가장 높은 종양 억제을을 나타냈으며 그 분자량은 47,000 dalton이었다. 그 구성성분은 82% 다당류, 8% 단백질 및 0.9% hexosamine 이었다. 그 다당류를 구성하는 단당류를 분석한 결과 glucose, galactose, mannose, fucose순으로 함유되어 있었다. 항암작용의 기전을 밝히기 위하여 마우스의 면역에 미치는 항암 성분의 영향을 연구한 결과 활성화된 마크로파지에서 분비되는 superoxide anion 양을 1.6배로, 그리고 hemolytic plaque assay에서의 용혈반 형성 세포수를 6배로 각각 증가시켰다. 이 결과로 항암작용은 고형암에 직접적으로 세포독성을 나타내는 것이 아니라, 면역세포를 활성화 혹은 강화시키는 작용임을 알 수 있다.

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Effects of antitumor polysaccharides from albizza julibrissin on immune function

  • Moon, Chang-Kiu;Lee, Byeong-Gon;Lee, Soo-Hwan;Kang, Tak-Lim
    • Archives of Pharmacal Research
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    • 제8권4호
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    • pp.277-282
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    • 1985
  • Polysaccharide fractions were prepared from Albizzia julibrissin by different extraction schedules. The fractions obtained were designated as PS-I and PS-II, respectively. Further purification of PS-I by Sephadex G-200 column chromatography gave two subfractions, Alju A and alju B, Each fraction showed marked antitumor activity against sarcoma 180 solid form but not ascite form. PS-I and PS-II increased delayed hypersensitivity in normal and tumor bearing mice. PS-I treatment led to moderate restoration of the suppressed antibody production in tumor bearing mice.

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솔잎, 돌나물, 톳, 메밀, 깻잎 등 5가지 혼합 열수 추출물의 면역 활성 효과 (Studies on the Effects of Water Extract from Mixture of Pine Needles, Sedum sarmentosum Bunge, Hijkiaorme, Buckwheat and Perlla Leaves on the Immune Function Activation)

  • 류혜숙;김현숙
    • 한국식품영양학회지
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    • 제21권3호
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    • pp.269-274
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    • 2008
  • Plants have long been used as a food source in Korea. In this study, we investigated the combined immunomodulative effects of a water extract mixture of(pine needles, Sedum sarmentosum Bunge, hijkiaorme, buckwheat and Peril a leaves) on Balb/c mice $7{\sim}8$ weeks old. The mice were fed a chow diet ad libitum and the plant extract was orally administered every other day for four weeks at two different concentrations(50 and 500 mg/kg BW). After preparing the single-cell suspension, splenocyte proliferation was determined by the MTT(3-[4,5-di-methylthiazol-2-y]-2,5-diphenyl terazolium bromide) assay. After 48hrs of incubation with the mitogens(ConA or LPS) splenocyte from the mice groups administered 50 and 500 mg/kg BW of the plant extract showed a significant increased in proliferation compared to the control group. A hemolytic plague forming cell assay was used to indicate antibody production against sheep red blood cells(SRBC). The number of antibody-secreting cells T-dependent antigen. The result of this study suggest that supplementation with this plant extract may regulate immune function by increasing splenocyte proliferation and the number of plaque forming cells.

우황이 생쥐의 면역반응에 미치는 영향 (Effects of the Administration of Bezoar Bovis on Immune Responses of Mice)

  • 손은화;박재현;김경란;김병오;이동권;표석능
    • 생약학회지
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    • 제29권1호
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    • pp.48-55
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    • 1998
  • The effects of Bezoar Bovis on immune responses of ICR Mice were studied. In the present study, the Jerne hemolytic plaque assay (PFC) was used to evaluate the humoral immune response to sheep red blood cells, and macrophage regulatory function was measured by quantitating the production of molecules secreted by macrophages. Bezoar Bovis was given in single daily p.o doses for short term (1, 2 days) or long term (7, 14 days). PFC response for taken 3 days after short term exposure to Bezoar Bovis was increased. The production of nitirc oxide in macrophages was also stimulated. In contrast, long term exposure to Bezoar Bovis resulted in inhibitory effect of Bezoar Bovis on nitirc oxide, $TNF-{\alpha}$and IL-6 production in macrophages. These findings suggest that treatment with Bezoar Bovis may result in differential immunological effect depending on treatment schedule.

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Immunomodulating activities of water extract from xanthium strumarium 2

  • Moon, Eun-Yi;Park, Seung-Yong;Ahn, Mee-Ja;Ahn, Jong-Woong;Zee, Ok-Pyo;Park, Eun-kyue
    • Archives of Pharmacal Research
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    • 제14권3호
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    • pp.217-224
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    • 1991
  • One of water and/or methanol extracts from 14 herbal deugs which were screened using murine splenocytes showed immunosuppressive activities previously. After water extract from Xanthium strumarium was treated with chloroform. $100 \mu{g/ml}$ of water layer (XS-WCI) has very strong immunosimulating activities tested by $^3H$-thmidine incorporation (control as $100 \mu{g/ml}$, 26345 cpm was 69515 cpm). MLR also appears to be simulated strongly (control vs $100 \mu{g/ml}$, 4962 cpm vs 78688 cpm). When $100 \mu{g/ml}$ of XS-WCI and $0.8 \mu{g/ml}$ of concanavalin a (ConA) were added. more $^3H$-thymidine were incorporated significantly, compared with $0.8 \mu{g/ml}$ of ConA only. In contrast with ConA. results from $5 \mu{/ml}$ of lipopolysaccharide (LPS) and $100 \mu{g/ml}$ of XS-WCI were not different. compared with $5\mu{/ml}$of LPS only. These results indicated the responses of XS-WCI to B cell and T cell may be different. XS-WCI was injected intraperitoneally (10 mg/kg. 50mg/kg/ 100 mg/kg) for 4 days or 10 days and tested secretion of IgM or IgG by direct and indirect hemolytic plaque-forming cell assays, respectively. Numbers of hemolytic plaques for both IgM and IgG were increased significantly. Especially, secretion of IgGs was increased more than 10 times. After administration of XS-WCI for 7 days (50 mg/kg. 100 mg/kg) splenomegaly deu to graft vs host reaction was observed. Human lymhocytes separated from whole blood by Ficoll-Hypaque method were also proliferated after treatment of $10 \mu{g/ml}$ and $50 \mu{g/ml}$ of XS-WCI. As seen in murine lymphocytes, human lymphocyte proliferation was increased synergistically after treatment with both of XS-WCI and phytohemagglutinin (PHA). It appears that XS-WCI may have potential immunosimulating activities and that it remains to be purified further for isolation of active components.

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갓버섯의 항균 분석 (Antitumor Components of the Cultured Mycelia of Lepiota procera)

  • 김병각;심미자;김옥남;김하원;최응칠
    • 한국식품위생안전성학회지
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    • 제4권2호
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    • pp.109-118
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    • 1989
  • 한국산 담자균류인 갓버섯 Lepiota procera의 균사를 액내 배양하여 항암성분인 단백성 다당체를 분리하였다. 이 성분은 DEAE-Sephadex A-50 이온교환수지와 Sepharose-4B gel Filtration을 이용하여 정제하여 Fraction C1을 얻었으며 이 Fr, Cr은 단백질과 다당체로 구성되어 있으며 항암 효과는 10mg/kg/day 투여군에서 64%의 저지유을 나타내었다. 이러한 항암작용의 기전을 밝히기 위한 연구의 일환으로 면역에 미치는 영향을 실험함 결과 이 단백다당체는 용혈반형성 세포수를 증가시켰으며, 저하된 지연성 과민반응을 회복시켰을 뿐만 아니라, carrageenan 투여에 의해 억제된 면역능을 다시 증강시켰음을 알 수 있었다. 이러한 결과들은 이 버섯의 항암작용이 세포독성에 의한 것이 아니라 종양에 대한 면역능을 강화시켜 발휘됨을 제시하고 있다.

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백작약 조다당분획에 의한 B 세포 증식의 특성 (Characteristics of B cell proliferation by polysaccharide fraction of Paeonia japonica miyabe)

  • 박혜란;함연호;이성태;백상기;조성기
    • IMMUNE NETWORK
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    • 제1권2호
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    • pp.126-134
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    • 2001
  • Background : Paeonia japonica Miyabe is a medicinal plant which has been widely used as a component of blood-building decoctions (Chinese medicinal concept : Bu-Xie). The immunopharmacological characteristics of the extract of Paeonia japonica (PJ) were investigated. Methods : The effects of fractions of PJ extract on lymphocyte proliferation were measured by $H^3$-thymidine incorporation assay. The proliferated lymphocyte subsets were analyzed in flow cytometry. The subset cell populations of spleen cells were separated by magnetic cell separation system, and their proliferation by the extract were investigated. The effect of the extract on antibody production was determined in mice challenged with sheep red blood cells (SRBC) using hemolytic plaque forming cell assay. Results : Spleen cells were proliferated by water extract of PJ. Polysaccharide fraction (PJ-P) of the extract was most active in the proliferation. It was found in flow cytometry that the lymphocyte subset proliferated by PJ-P was B cell population. Among the separated subset cell populations, T cell-depleted cell population and macrophage-depleted cell population were most proliferated by PJ-P. However, positively selected populations of B cells and T cells were not proliferated by PJ-P. These results indicate that B cell proliferation by PJ-P may require the assistance of macrophages or T cells. These results suggest that firstly PJ-P may stimulate macrophages or T cells, and then B cells are activated. The number of antibody-secreting cells was increased by administration of PJ-P in mice immunized with SRBC as a T-dependent antigen. Conclusion : These results suggest that macrophages and accessory cells are directly activated by PJ-P and then helper T cells and B cells are indirectly activated. As the results, immune responses might be coordinately improved. In conclusion, PJ-P, a polysaccharide of P. japonica, may be a characteristic immunostimulator, which is analogous to polysaccharides such as lentinan, PSK and ginsan.

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