• The Korean Journal of Physiology and Pharmacology
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• v.10 no.1
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• pp.19-24
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• 2006

The intermediate conductance $Ca^{2+}-activated$ $K^+$ channels (SK4, IKCa1) are present in lymphocytes, and their membrane expression is upregulated by various immunological stimuli. In this study, the activity of SK4 was compared between Bal-17 and WEHI-231 cell lines which represent mature and immature stages of murine B lymphocytes, respectively. The whole-cell patch clamp with high-$Ca^{2+}$ ($0.8{\mu}M$) KCl pipette solution revealed a voltage-independent $K^+$ current that was blocked by clotrimazole (1 mM), an SK4 blocker. The expression of mRNAs for SK4 was confirmed in both Bal-17 and WEHI-231 cells. The density of clotrimazole-sensitive SK4 current was significantly larger in Bal-17 than WEHI-231 cells ($-11.4{\pm}3.1$ Vs. $-5.7{\pm}1.15$ pA/pF). Also, the chronic stimulation of B cell receptors (BCR) by BCR-ligation (anti-IgM Ab, $3{\mu}g$/ml, 8∼12 h) significantly upregulated the amplitude of clotrimazolesensitive current from $-11.4{\pm}3.1$ to $-53.1{\pm}8.6$ pA/pF in Bal-17 cells. In WEHI-231 cells, the effect of BCR-ligation was significantly small ($-5.7{\pm}1.15$ to $-9.0{\pm}1.00$ pA/pF). The differential expression and regulation by BCR-ligation might reflect functional changes in the maturation of B lymphocytes.

• YAKHAK HOEJI
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• v.37 no.3
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• pp.254-261
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• 1993

Developing new substance for immunosuppressor or immunomodulator from natural products is extremely important in the present biomedicine. In this paper, we focused our efforts on the mitogenicity and immunochemical properties of the two lectins (LCC-I, LCC-II) obtained from marine animal Lunella coronata coreensis. Immunochemical techniques were employed to elucidate the structural and/or functional similarities between the LCC lectins. Molecular weight of the LCC lectins, LCC-I and LCC-II were estimated to be around 60 KD and 66-70 KD, respectively. LCC lectins were mitogens for murine splenic lymphocytes, and the optimum mitogenic doses were 31.25 $\mu\textrm{g}$/ml and 3.91 $\mu\textrm{g}$/ml, respectively. LCC-II lectin was a good mitogen toward human peripheral lymphocytes at a concentration about 31.25 $\mu\textrm{g}$/ml.

• YAKHAK HOEJI
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• v.39 no.3
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• pp.252-259
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• 1995

Isolation, purification and characterization of biophysicochemical properties of the three new lectins, MLA-I, MLA-II, MLA-III from the hemolymph of Meretrix lusoria have been reported previously. A series of immunochemical studies were investigated in this work. The three lectins were revealed as having partial identity each other by immunodiffusim and immunoelectrophoresis. These results suggest that MLA lectins are isolectins having similar biophysicochemical properties. Particularly, MLA-I proved to be a potent mitogen for murine splenic as well as human peripheral lymphocytes, and the optimum mitogenic dose were 62.5 and 1.95 $\mu\textrm{g}$/ml, respectively.

• Journal of Applied Biological Chemistry
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• v.48 no.4
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• pp.170-177
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• 2005

Oriental medicinal herb extracts (OHE) showing anticancer activities were investigated for effectiveness as antiviral drugs. Infection of MuLV to cell line resulted in formation of giant syncytia. Number of giant syncytia in culture treated with OHE decreased by 40% compared to that of non-OHE-treated cell culture. To determine OHE effects on progeny release, RT-PCR was performed. In vivo animal studies demonstrated effectiveness of OHE as antiviral drug when administered orally. After OHE administration, viral cytopathic effects decreased. Infected mice showed splenomegaly and over-proliferation of lymphocytes with decreased CD4+ cell counts. These symptoms decreased in OHE-treated mice, indicating OHE maybe useful therapeutics against MuLV/MAIDS as Human Immunodeficiency Virus (HIV)/AIDS animal model. Results show XC plaque assay and in vivo MAIDS model using MuLV are suitable tools for screening anti-retroviral drug candidates.

• YAKHAK HOEJI
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• v.42 no.4
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• pp.467-471
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• 1998

Numerous efforts have been made to isolate immunologically active component from Mori Cortex Radicis, since it has been used in the treatment of bronchial asthma, and immune dis order in human. Recently, we reported the purification of an anti-allergic component of the Mori Cortex Radicis. Among the fractions we prepared in the previous study, a fraction was active in the proliferation of murine lymphocytes. The active component (HHM 3-1) was elucidated as a polysaccharade with a small amount of lignin. When it was subjected to MALDI-MS by using 3-hydroxypicolinic acid as a matrix, the molecular weight of the component was estimated as 792688.2dalton. Total hexose and protein content of the component were estimated as 62.6% and 0.51%, respectively and it was composed mainly of glucose, galactose and mannose. The remaining part of the component was estimated as ligin because of the characteristic functional groups in IR and UV spectra. Concomitant treatment of HHM 3-1 with known mitogens synergistically increased the proliferation of B-cells and T-cells.

• Archives of Pharmacal Research
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• v.20 no.2
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• pp.128-137
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• 1997

The effects of cylindan, a polysaccharide isolated from the basidiocarps of Agrocybe cylindracea, on murine sarcoma 180 tumor and murine immune cells were examined after intraperitoneal administration. Cylindan exhibited a marked life extension effect in mice against ascite forms of sarcoma 180 and Lewis lung carcinoma at a dose of 50 mg/kg/day, although it did not show any direct cytotoxicity against sarcoma 180, X5563, and MM46 murine tumor cells. Cylindan increased numbers of bone marrow stem cells as well as peritoneal exudate cells in flow cytometry using monoclonal antibodies. The tumor bearing mice group apparently showed the increase of macrophages and cytotoxic T lymphocytes in mouse spleen cells during the early stage of tumor growth. But during the later stage, the control group decreased immune cells and cylindan restored the decreased immune cells in the tumor bearing mice to the normal level. In non-specific immune response, cylindan stimulated the bacterial phagocytosis and acid phosphatase production in macrophages. It also activated components of the alternative complement pathway and natural killer activity against YAC-1 lymphoma. In number of plasma cells as token of stimulation of the differentiation of B lymphocytes. In cellular immunity, cylindan restored the depressed response of delayed type hypersensitivity in the tumor bearing mice to 60% of the normal level and increased the interleukin-2 (IL-2) responsiveness in the IL-2 dependent CTLL-2 cells. These results suggest that cylindan did not show direct cytotoxic effects on tumor cells but restored the decreased immune response of the tumor bearing mice.

• Archives of Pharmacal Research
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• v.23 no.3
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• pp.240-242
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• 2000

The immunomodulating activity of a polysaccharide isolated from Morus alba (PMA) root bark was examined in murine splenic lymphocytes. PMA enhanced proliferation of splenic lymphocytes in a synergistic manner in the presence of mitogens. However, PMA suppressed pri-may IgM antibody production from B cells, which was activated with lipopolysaccharide, a polyclonal activator, or immunized with a T-cell dependent antigen sheep red blood cells. Our observations showed that the immunomodulating activity of PMA increased lymphocyte proliferation and that PMA decreased antibody production from B cells, which was distinct from those of other plant-originated polysaccharides.

• 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.

• BMB Reports
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• v.28 no.6
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• pp.556-561
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• 1995

The purpose of this study was to establish an in vitro culture method of tumor-specific T cells, and determine the efficacy of the cultured tumor-specific cytotoxic T-lymphocytes (CTL) as an agent of anti-tumor immunotherapy against a murine lymphoma, TIMI.4. Tumor-specific T-lymphocytes derived from C57BL/6 mice (thy-1.2) immune to TIMI.4 were activated by in vitro stimulation with the irradiated TIMI.4 cells, and expanded by restimulation with TIMI.4 in the presence of the concanavalin A-stimulated rat spleen culture supernatant, and splenic antigen-presenting cells. In vitro restimulation enhanced markedly the proportion of $CD8^+$, a predominant surface marker of CTL and the cytotoxic activity in the cultured immune T cell population. The resulting TIMI.4-specific T cells were adoptively transferred into nude mice. The tumor cells residing in the host after 7 days of adoptive transfer to B6.PL (thy-1.1) mice were quantified by use of an antibody directed to the thy-1.2 allele. The TIMI.4 cells in the recipient nude mice were decreased in a dose-dependent manner. Anti-tumor activity of the TIMI.4-specific T cells was also demonstrated by a survival test, where the tumor-bearing nu/nu mice which received the activated T-cells survived about 30% longer than the control mice which received the tumor cells alone. These suggest that adoptive transfer of TIMI.4-specific T cells could be a candidate for effective therapy of the murine lymphoma.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.137.2-137.2
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• 2003

Dendritic cells (DCs) are known to professional antigen presenting cell (APC). Due to the role as an effective activator of cytotoxic T Lymphocytes by expressing MHC, adhesion and co-stimulatory molecules, DCs are now widely recognized to play an important role in the immune responses to tumors.We investigated the effect of cultured DCs in murine melanoma pulmonary metastasis model. To follow the metastasis protocol, syngenic melanoma cells were inoculated intra-venously into the mouse (B16F10 into the C57BL/6)8 days prior to the first DC injection (1$\times$106 DCs/ mouse, i.p.) and the autologous tumor cell lysate pulsed-DCs were injected as a therapeutic module twice in two weeks. (omitted)