• Title/Summary/Keyword: Murine lymphocytes

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Isolation and Characterization of Lectin from Viscum coloratum (겨우사리 중의 렉틴성분 분리 및 특성)

  • Park, Won-Bong;Kim, Hee-Sook
    • YAKHAK HOEJI
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    • v.38 no.4
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    • pp.418-424
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    • 1994
  • Lectin from mistletoe (Viscum coloratum) were obtained by salt fractionation, gel filtration and anion exchange chromatography. A molecular weight of about 60,000D has been determined by SDS-PAGE and two basic subunits which have molecular weights of 33,000D and 28,000D are linked by a disulfide bond. The partially purified mistletoe lectins agglutinated human B erythrocytes. Agglutinating activity was relatively stable at varied pH $(3.77{\sim}8.71)$ and at temperature range of $0{\sim}40^{\circ}C$ and not affected by 9 metal ions. Galactose, lactose and N-acetyl-D- galactosamine inhibited agglutinating activity of lectin. Lectin from mistletoe was more mitogenic to murine lymphocytes than concanavalin A.

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Studies on Lectins from Marine Animal Chlorostoma argyrostoma turbinatum (해양동물 구멍밤고둥의 렉틴 성분 연구)

  • Chung, See-Ryun;Choi, Il-Shik;Jeune, Kyung-Hee
    • Korean Journal of Pharmacognosy
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    • v.25 no.2
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    • pp.121-131
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    • 1994
  • Two kinds of new lectins, CATL-I and CATL-II, were partially purified from the intestine of Chlorostoma argyrostoma turbunatum by physical saline extraction, salt fractionation, ion exchange chromatography and gel filtration. CATL-I and CATL-II were purified 39.4 and 15.8 fold with a yield of 8.8 and 7.4%, respectively. On polyacrylamide gel electrophoresis, CATL-I demonstrated one major and one minor bands. This lectin agglutinated human and other animal erythrocytes nonspecifically and also agglutinated murine splenic lymphocytes. Carbohydrate specificity of the lectins was determined by inhibition of the agglutinability by methyl-${\alpha}-_D$-galactopyranoside and $_L-rhamnose$ at a final concentration of 6 mM. The lectins contained relatively high amounts of acidic amino acids, but the contents of sulfur containing amino acids were very low or was not estimated. Immunochemical studies were carried out to identify some properties of marine animal lectins.

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Immunosuppressive Activity of Cepacidine A, a Novel Antifungal Antibiotic Produced by Pseudomonas cepacia

  • LEE, CHUL-HOON;JUNG-WOO SUH;YOUL-HEE CHO
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.672-674
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    • 1999
  • Cepacidine A was first identified as a novel antifungal antibiotic which was isolated from the culture broth of Pseudomonas cepacia AF200l. It showed a potent in vitro antifungal activity against various pathogenic fungi, but did not show any activity against bacteria. Recently, the immunosuppressive action of cepacidine A was discovered using an in vitro screening system involving inhibition of the proliferation of murine lymphocytes stimulated by 2 mitogens, and also by in vivo mouse models involving inhibition of delayed type hypersensitivity and SRBC hemagglutination. Cepacidine A showed a significant activity of cellular immunosuppression (ED/sub 50/) at concentration levels of 1-3 ㎎/㎏, i.p.. Unfortunately, the delayed toxicity at a dose of above 3 ㎎/㎏ i.p. was apparent.

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Effect of Polysaccharide Extracted from Panax ginseng on Murine Hematopoiesis (인삼 다당체가 생쥐의 조혈과정에 미치는 영향)

  • 송지영;이세윤;정인성;윤연숙
    • Journal of Ginseng Research
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    • v.25 no.2
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    • pp.63-67
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    • 2001
  • We previously reported that acidic polysaccharide from Panax ginseng induced the proliferation lymphocytes and the generation of activated killer cells. Here we found that polysaccharide (PG-75) precipitated with 75% EtOH from water extract of Panax ginseng also has both in vitron and in vivo hematopoietic activities. In vitro studied with bone marrow cells from BALB/c mouse revealed that PG-75 had direct effect on hematopoietic colony-forming cell(CFC) growth, increased granulocyte macrophage-colony forming cell numbers by 1.59 fold over than non-treated. the ability of PG-75 to modulate hematopoiesis in vivo was evaluated the bone marrow and spleen celluarity, granulocyte-macrophage progenitor cells. BALB/c female mice were administered G-75 intraperitoneally, PG-75 was found to significantly increase the number of BM cells, spleen cells, GM-CFU on 3 hours after injection. PG-75 was also able to induce significant augmentation of GM-CSF and IFN-${\gamma}$, production in sera. These studies illustrate than PG-75 has hematopoietic activities and that this agent may be useful in the prevention and/or treatment of radio- or chemotherapy-associated myelosuppression.

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Enhancement of Immune Responses by a Water Soluble Proteoglycan, Lepidan from Lentinus lepideus (잣버섯 균사체로부터 분리한 수용성 단백다당체 Lepidan의 면역 증가 작용)

  • 진미림;정규선
    • YAKHAK HOEJI
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    • v.43 no.5
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    • pp.635-641
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    • 1999
  • In this study, we investigated the immunopotent activities of lepidan, a water soluble proteoglycan from Lentinus lepideus. To examine whether lepidan may affect nonspecific immune responses, we determined the effect on the production of nitric oxide (NO). Lepidan effectively increased the NO production in IFN-${\gamma}$ and LPS triggered RAW 264.7 cells. To further demonstrate the evidence that lepidan activates various immune cells, we determined the alkaline phosphatase activity, plaque-forming cell number and secretion of interleukine-4 (IL-4) and granulocyte/macrophage-colony stimulating factor (GM-CSF) after lepidan treatment in murine splenocytes. The results showed that lepidan increased alkaline phosphatase activity and the number of plaque-forming cells, which indicates that lepidan can lead B lymphocytes to late stage of differentiation. Also, when the splenocytes were cultured with lepidan for 48 hr, the level of IL-4 and GM-CSF in the supernatant dramatically increased. Taken together, these data suggest that lepidan is a biological response modulator that is able to activate immune responses.

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Ameliorative effects of Glechoma hederacea var. longituba extract on oxazolone-induced atopic dermatitis-like skin lesions (긴병꽃풀 추출물의 oxazolone-유도 아토피 피부염 개선 효과)

  • Dae Yong Kim
    • Journal of Convergence Korean Medicine
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    • v.5 no.2
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    • pp.17-22
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    • 2023
  • Objectives: This study aimed to investigate the anti-atopic properties of Glechoma hederacea var. longituba extracts (GHE) in murine atopic dermatitis model. Methods: BALB/c mouse ear stimulated with oxazolone (OX) for 4 weeks, then 1% GHE was topically applied every two days for 3 weeks to mouse. Ear thickness was measured by a digital thickness gauge. The ears tissues were collected and subjected to hematoxylin-eosin (H&E) and toluidine blue (TB) staining. Results: Treatment with GHE successfully alleviated the symptoms of atopic dermatitis, such as erythema, horny substance, and swelling. The infiltration of lymphocytes and mast cells were significantly reduced following GHE treatment. Conclusion: Taken together, our results showed that GHE possessed the potential to be a novel immunomodulatory drug against atopic dermatitis.

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The Effects of Alkaloid Fraction of Korean Ginseng on the Radiation-Induced DNA Strand Breaks (방사선 조사에 의한 DNA Double Strand Breaks의 생성 및 회복에 미치는 인삼 알칼로이드 분획의 효과)

  • Cho Chul Koo;Kim Tae Hwan;Yoo Seong Yul;Koh Kyoung Hwan;Kim Mi Sook;Kim Jeong Hee;Kim Seong Ho;Yoon Hyung Keun;Ji Young Hoon
    • Radiation Oncology Journal
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    • v.13 no.2
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    • pp.113-120
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    • 1995
  • Purpose : To investigate the effect of alkaloid fraction from Korean ginseng on radiation-induced DNA double strand breaks (dsb) formation and repair in murine lymphocytes Materials and Methods : We used the neutral filter elution technique to assay $^{60}Co\;{\gamma}$ ray-induced DNA double strand breaks formation and repair in C57BL/6 mouse spleen lymphocytes for evaluating the dose-response relationship in the presence of alkaloid fraction as a radioprotective agent. The lymphocytes were stimulated with Phytohemagglutinin (PHA, 2 u g/ml) to label $^3[H]-thymidine.$ Isotope-labelled lymphocytes in suspension were exposed to 100 Gy at $0^{\cdot}C$ in the alkaloid fraction-treated group and elution procedure was performed at PH 9.6. The extents of formation of radiation-induced DNA double strand breaks and repair were compared respectively via strand scission factor (SSF) and relative strand scission factor (RSSF). Results: Alkaloid fraction reduced the formation of double strand breaks with dose modification factor of 2 15, compared to control group Rejoining of DNA dsb appeared to take place via two components. The first fast component was completed within 20.4 minutes, but the second slow component was not completed until 220.2 minutes after irradiation. About $30\%$ of dsb formed by irradiation was ultimately unrejoined despite the administration of alkaloid fraction. The administration of alkaloid fraction had a great effect on the second slow component of repair; the half-time of fast component repair was not changed, but that of slow component was 621.8 minutes. Conclusion: Neutral filter elution assay Proved to be a very effective method to quantitate the extents of DNA dsb formation and its repair. By using this technique, we were able to evaluate the efficiency of alkaloid fraction from Korean ginseng as a valuable radioprotector. Alkaloid fraction can be used prophylactically to prevent or ameliorate the severe radiation damages in workers and neighbors around the atomic power plants. For more refined study, however, more advanced purification of alkaloid fraction wil be needed in the near future.

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Effective Antitumor Activity of a Recombinant Vaccinia Virus Expressing Murine Interleukin 4 (인터루킨-4를 발현하는 재조합 백시니아 바이러스에 의한 암성장의 억제)

  • Yoon, Kee-Jung;Jin, Ning-Yi;Kim, Sun-Young
    • The Journal of Korean Society of Virology
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    • v.28 no.1
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    • pp.71-78
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    • 1998
  • Vaccinia virus is the prototype orthopoxvirus that has been used as a vaccine strain for small pox. This virus has been used to express a variety of cellular and viral genes in mammalian cells at high levels. Interleukin-4 (IL-4) has been found to stimulate the proliferation of T cells and enhance the cytolytic activity of cytotoxic T lymphocytes. To test the immunotherapeutic potential of IL-4 delivered in vivo by poxvirus, a recombinant vaccinia virus expressing the murine IL-4 gene (RVVmIL-4) was constructed. A high level of IL-4 production was confirmed by infecting HeLa cells and measuring IL-4 in cell culture supernatant by ELISA. As a tumor model, two cell lines were used; the murine T leukemic line P388 and the murine breast cancer line TS/A. CDF1 mice were intraperitoneally inoculated with $1\;{\times}\;10^5$ cells of P388. Mice were injected at the same site with $5\;{\times}\;10^5\;PFU$ of recombinant vaccinia virus; first, 3 days after the injection of tumor cells and thereafter once every week for 3 weeks. Intraperitoneal injections of RVVmIL-4 significantly prolonged the survival time of mice inoculated with tumor cells. All mice injected with RVVmIL-4 remained alive for 30 days after the postinoculation of tumor cells, while 100% and 70% of the animals injected with saline or wild type vaccinia virus died, respectively. In another tumor model using TS/A, tumor was established by subcutaneously inoculating $2{\times}10^5$ tumor cells to BALB/c mice. After tumor formation was confirmed on day 4 in all mice, $5\;{\times}\;10^6\;PFU$ of RVVmIL-4 was inoculated subcutaneously three times, once every week for 3 weeks. The TS/A tumor was eradicated in two of the nine mice. Seven of the nine mice treated with RVVmIL-4 developed a tumor, but tumor growth was significantly delayed compared to those treated with saline or wild type vaccinia virus. These results indicate that recombinant vaccinia viruses may be used as a convenient tool for delivering immunomodulator genes to a variety of tumors.

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Hemopoietic and Radioprotective Effects of Several Herbs on Murine Splenocytes and Bone Marrow Cells (수(數) 종(種) 한약재(韓藥材)가 생쥐의 골수(骨髓) 및 비장세포(脾臟細胞)의 조혈촉진(造血促進)과 방사선(放射線) 방어(防禦)에 미치는 영향)

  • Lee, Neung-Gi;Choe, Seung-Hun;An, Gyu-Seok
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.3 no.1
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    • pp.29-48
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    • 1997
  • Astragalus membranaceus Bunge (黃?), Atractylodes macrocephala Loidz (白朮), Angelica sinensis(Oliv) Kiels (當歸), Lycium chinense Mill (枸杞子), Ligustrum lucidum Ail (女貞子) are Herbs that are frequently used in a lot of prescriptions to reduce the side effects of anti-cancer therapies, especially like chemotherapy and radiotherapy. The radioprotective and hemopoietic effects of these Herbs on BALB/c splenocytes and bone marrow cells are measured. In order to evaluate the Hemopoietic effects, Thymidine uptakes and secretion of colony stimulating factors(CSFs) of splenocytes and myelocytes treated with herbs were measured. Radioprotective effects were accessed by the method of immunocompetence of murine lymphocytes cultured with herbs before irradiation and with LPS, ConA after irradiation. The results are as follows. 1. The orders of Proliferative effects of herbs on splenocytes were Atractylodes macrocephala Loidz (白朮), the higest of all, Astragalus membranaceus Bunge (黃?), Angelica sinensis(Oliv) Kiels (當歸). Lycium chinense Mill (枸杞子) and Ligustrum lucidum Ait (女貞子), the lowest of all. At Optimal concentration, the proliferation ratios of herb-treated splenocytes compared to non-treated ones were like these. Atractylodes macrocephala Loidz (白朮) 44.3, Astragalus membranaceus Bunge (黃?) 17.7, Angelica sinensis(Oliv) Kiels (當歸) 10, Lycium chinense Mill (枸杞子) 6.4, Ligustrum lucidum Ait (女貞子) 2.0.(p<0.05) 2. When splenocytes were cultured during different periods, Atractylodes. macrocephala Loidz (白朮) and Astragalus membranaceus Bunge (黃?) showed the higest proliferation on 3th day, Angelica sinensis(Oliv) Kiels (當歸), Lycium chinense Mill (枸杞子) on 4th day,(p<0.05) and Ligustrum lucidum Ait (女貞子) until 5th day but with no significant increase. 3. The orders of Proliferative effects of herbs on Bone Marrow(BM) cells were Atractylodes macrocephala Loidz (白朮), the higest of all, Astragalus memhranaceus Bunge (黃?), Angelica sinensis(Oliv) Kiels (當歸), Lycium chinense Mill (枸杞子) and Ligustrum lucidum Ait (女貞子), the lowest of all too. At Optimal concentration, the proliferation ratios of herb-treated BM cells compared to non-treated ones were like these. Atractylodes macrocephala Loidz (白朮) 21.7, Astragalus membranaceus Bunge (黃?) 9.9, Angelica sinensis(Oliv) Kiels (當歸) 4.9, Lycium chinense Mill (枸杞子) 2.3, Ligustrum lucidum Ait (女貞子) 1.4(p<0.05). 4. The secretion ratio of colony stimulating factors(CSFs) of each herb-treated group, compared to control, was Atractylodes macrocephala Loidz (白朮) 9.4, Astragalus membranaceus Bunge (黃?) 9.0, Angelica sinensis(Oliv) Kiels (當歸) 4.4, Lycium chinense Mill (枸杞子) 3.8 (p<0.05) but no significant increase in Ligustrum lucidum Ait (女貞子). 5. The mitogen(ConA, LPS) stimulated-lymphocytes cultured with each herb before irradiation of 1-3 Gy showed more enhanced proliferation than control(p<0.05). When compared to each non-irradiated group of all groups, the orders of percentage increase of irradiated group were Atractylodes macrocephala Loidz (白朮), the higest of all, Astragalus membranaceus Bunge (黃?), Angelica sinensis(Oliv) Kiels (當歸), Ligustrum lucidum Ait (女貞子), Lycium chinense Mill (枸杞子). Each percentage showed significant enhancement compared to control group(p<0.05). According to the results, Atractylcdes macrocephala Loidz (白朮), Astragalus membranaceus Bunge (黃?) are suggested to be the most effective hemopoietic and radioprotective herbs, and Angelica sinensis(Oliv) Kiels (當歸), Lycium chinense Mill (枸杞子) the next, but Ligustrum lucidum Ait (女貞子) showed lower effects than expected.

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