• Title/Summary/Keyword: Hemolytic plaque assay

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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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    • v.5 no.2
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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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    • v.16 no.4
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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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Studies on Constituents of Higher Fungi of Korea (LXXIII) Antitumor Components of the Cultured Mycelia of Paxillus atrotomentosus (한국산 고등 균류의 성분 연구(제73보) -좀우단버섯 배양 균사체의 항암 성분-)

  • Kwag, Sang-Deok;Bok, Jin-Woo;Hyun, Jin-Won;Choi, Eung-Chil;Kim, Byong-Kak
    • The Korean Journal of Mycology
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    • v.20 no.3
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    • pp.240-251
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    • 1992
  • To find antitumor components from higher fungi, the cultured mycelia of Paxillus atrotomentosus were extracted with hot water. The water soluble fraction was purified and separated by DEAE-cellulose ion exchange chromatography and Sepharose CL-4B gel filtration method. The separated fractions(Fr.) were designated CR A, B, C and D. Fr. A showed the highest inhibition ratio of 68.51% among the five tractions at a dose of 20 mg/kg/day. When Fr. A was examined for immunopotentiation activity, it increased the amount of the superoxide anion from activated macrophages to 1.1 fold and the number of plaques in hemolytic plaque assay to 2.3 fold, respectively. Otherwise, it did not show direct cytotoxity in sarcoma 180. Delayed type hypersensitiyity reaction showed that the decreased footpad swelling of tumor-hearing was restored to the normal. These results indicate that antitumor activity was exerted through immunopotentiation. Its chemical analysis showed 86.36% polysaccharide, 1.52% protein and 1.64% hexosamine. The polysaccharide consisted of fucose, galactose, glucose, mannose and xylose. This component was named paxillan.

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

  • Hyun, Jin-Won;Choi, Eung-Chil;Kim, Byong-Kak
    • The Korean Journal of Mycology
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    • v.18 no.2
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    • pp.58-69
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    • 1990
  • ABSTRACT: In order to find physiologically active components from higher fungi, hot-water soluble components were extracted from the basidiocarps of Ganoderma lucidum. The extract was purified and separated by DEAE cellulose ion exchange chromatography and Sepharose CL-4B gel filtration method. The separated fractions were designated CR, IN, IA, GL and GH. Fraction GL showed the highest antitumor activity among the fractions and its molecular weight was found to be 47 KD. The tumor inhibition ratio of Fr. GL was 81 % at the dose of peritoneal administration of 20 mg/kg/day for 10 days in mice. Chemical analysis of this fraction showed 82% polysaccharide, 8% protein and 0.9% hexosamine. The polysaccharide moiety consisted of 63% glucose, 27% galactose, 7% mannose and 3% fucose. Fraction IN was found to increase the amount of superoxide anion in activated macrophages to 1.6-fold and the number of plaques in hemolytic plaque assay to 6-fold, respectively. These results indicate that the antitumor activity was exerted through immunopotentiation, but not through direct cytotoxicity against the tumor.

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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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    • v.8 no.4
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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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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 (솔잎, 돌나물, 톳, 메밀, 깻잎 등 5가지 혼합 열수 추출물의 면역 활성 효과)

  • Ryu, Hye-Sook;Kim, Hyun-Sook
    • The Korean Journal of Food And Nutrition
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    • v.21 no.3
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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.

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

  • Son, Eun-Wha;Park, Jae-Hyun;Kim, Kyung-Ran;Kim, Byung-O;Rhee, Dong-Kwon;Pyo, Suhk-Neung
    • Korean Journal of Pharmacognosy
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    • v.29 no.1
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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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    • v.14 no.3
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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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Antitumor Components of the Cultured Mycelia of Lepiota procera (갓버섯의 항균 분석)

  • 김병각;심미자;김옥남;김하원;최응칠
    • Journal of Food Hygiene and Safety
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    • v.4 no.2
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    • pp.109-118
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    • 1989
  • To find antitumor components in the cultured mycelia of Lepiota procera, the proteinpolysaccharide obtained from the mycelia was subjected to DEAE - Sephadex A-50 column chromatography and Sepharose-4B gel filtration. Of the fractions, the purified Fraction C1 was named lepiotan and examined for antitumor activity against the solid form of sarcoma 180 in ICR mice. The inhibition ratio of lepiotan was 64% at the dose of 10 mg/kg/day for 10days. The chemical analysis of lepiotan showed 60% polysaccharide and 21 % protein. The polysaccharide moiety was found to be a heteromannoglucan which consisted of 46.3% glucose, 40.2% mannose and 11.0% fucose. When the antitumor component, Fraction A, was examined for immunopotentiation activity, it was found to increase the number of plaques in hemolytic plaque assay and to restore the immunity in the tumor-bearing mice up to 89.7% of the normal level. Also the antitumor acitivity was suppressed by the treatment with carrageenan, an antimacrophage agent. These results indicate that the antitumor activity was exerted through immunopotentiation, but not through direct cytotoxicity against the tumor.

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

  • Park, Hae-Ran;Ham, Yeon-Ho;Yee, Sung-Tae;Paik, Sang-Gi;Jo, Sung-Kee
    • IMMUNE NETWORK
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    • v.1 no.2
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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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