• IMMUNE NETWORK
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• 제9권6호
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• pp.274-284
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• 2009

Background: Swiprosin-1 was identified in human CD8+ lymphocytes, mature B cells and non-lymphonoid tissue. We have recently reported that swiprosin-1 is expressed in mast cells and up-regulated in both in vitro and in vivo. Methods: The expression of cytokines and swiprosin-1 were determined by by real time PCR and conventional PCR. Pharmacological inhibitors were treated to investigate potential mechanism of swiprosin-1 in mast cell activation. Actin content was evaluated by confocal microscopy and flow cytometry. Results: The swiprosin-1 augmented PMA/A23187-induced expression of cytokines and release of histamine. However, knock-down of swiprosin-1 showed only a modest effect on PMA/A23187-induced cytokine expression, suggesting that swiprosin-1 has gain-of-function characteristics. Swiprosin-1 was found in microvilli-like membrane protrusions and highly co-localized with F-actin. Importantly, either disruption of actin by cytochalasin B or inhibition of PI3 kinase, an enzyme involved in actin remodeling, by wortmannin blocked cytokine expression only in swiprosin-1-overexpressing cells. Conclusion: These results suggest that swiprosin-1 modulates mast cell activation potentially through actin regulation.

• The Korean Journal of Physiology and Pharmacology
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• 제9권1호
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• pp.55-62
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• 2005

Very little research has been carried out on safflower seed for the prevention and treatment of the bone deficiency diseases, including osteoporosis, which are supported by scientific evidences. In the present study, $3{\mu}l$ of 0.1% dried crude extract or $2{\mu}l$ of 0.1% dried aqueous fraction were shown to significantly accelerate the rate of differentiation of osteoblast. Also, the crude extract and aqueous fraction increased the $[Ca^{2+}]_i$ of the cultured osteoblast cells: $3{\mu}l$ of 0.1% dried crude extract and $2{\mu}l$ of 0.1% dried aqueous fraction significantly increased the $[Ca^{2+}]_i$ of the cultured osteoblast cells ($8{\times}10^{-4}$) to the extent that it deserves a considerable attention. Furthermore, the crude extract and aqueous fraction increased the $[Ca^{2+}]_i$ of the cultured osteoblast cells, and $300{\mu}M$ $Cd^{2+}$, specific calcium channel blocker, completely blocked the increase. Therefore, the increased $[Ca^{2+}]_i$ of the cultured osteoblast cells by safflower seed component continued to activate calcium channel.

• IMMUNE NETWORK
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• 제6권2호
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• pp.102-110
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• 2006

Background: Dendritic cells (DC) are professional antigen-presenting cells in the immune system and can induce T cell response against virus infections, microbial pathogens, and tumors. Therefore, immunization using DC loaded with tumor-associated antigens (TAAs) is a powerful method of inducing anti-tumor immunity. For induction of effective anti-tumor immunity, antigens should be efficiently introduced into DC and presented on MHC class I molecules at high levels to activate antigen-specific $CD8^+$ T cells. We have been exploring methods for loading exogenous antigens into APC with high efficiency of Ag presentation. In this study, we tested the effect of the cationic liposome (Lipofectin) for transferring and loading exogenous model antigen (OVA protein) into BM-DC. Methods: Bone marrow-derived DC (EM-DC) were incubated with OVA-Lipofectin complexes and then co-cultured with B3Z cells. B3Z activation, which is expressed as the amount of ${\beta}$-galactosidase induced by TCR stimulation, was determined by an enzymatic assay using ${\beta}$-gal assay system. C57BL/6 mice were immunized with OVA-pulsed DC to monitor the in vivo vaccination effect. After vaccination, mice were inoculated with EG7-OVA tumor cells. Results: BM-DC pulsed with OVA-Lipofectin complexes showed more efficient presentation of OVA-peptide on MHC class I molecules than soluble OVA-pulsed DC. OVA-Lipofectin complexes-pulsed DC pretreated with an inhibitor of MHC class I-mediated antigen presentation, brefeldin A, showed reduced ability in presenting OVA peptide on their surface MHC class I molecules. Finally, immunization of OVA-Lipofectin complexes-pulsed DC protected mice against subsequent tumor challenge. Conclusion: Our data provide evidence that antigen-loading into DC using Lipofectin can promote MHC class I- restricted antigen presentation. Therefore, antigen-loading into DC using Lipofectin can be one of several useful tools for achieving efficient induction of antigen-specific immunity in DC-based immunotherapy.

• 대한예방한의학회지
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• 제5권1호
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• pp.144-150
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• 2001

The cytotoxic activity of Cratalariae sessiliflorae on cultured NIH 3T3 fibroblasts and human oral epithelioid carcinoma cells (KB) were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide (MTT) colorimetric method These fractions of methanol extract of Cratalariae sessiliflorae showed inhibitory effect in vitro in the milligram range against KB cell lines. In general, the antitumor activities of these fractions were does-dependent over the milligram range. The comparison of IC50 values of these fractions in tumor cell lines showed that their susceptibility to these fractions decrease in the following order: Fr. 4> Fr. 6> Fr. 10> Fr. 2> Fr. 11> Fr. 3> Fr. 8> Fr. 7> Fr. 9> Fr. 1> Fr. 5 by the MTT assay. These fractions were tested for their cytotoxic effects on NIH 3T3 fibroblasts using MTT assay. They exhibited potent cytotoxic activities in vitro in the milligram range against NIH 3T3 fibroblasts. In general, the cytotoxic activities of these fractions were does-dependent over the milligram range. The comparison of CD50 values of these fractions in NIH 313 fibroblasts shows that their susceptibility to these fractions in decrease the following order: Fr. 10> Fr. 9> Fr. 2 = Fr. 4> Fr. 8> Fr. 11> Fr. 1 = Fr. 7> Fr. 3> Fr. 5 = Fr. 6 by the MTT assay. These results suggests that fraction 5 has the most growth - inhibitory activity against KB cell lines.

• Tuberculosis and Respiratory Diseases
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• 제52권2호
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• pp.117-127
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• 2002

배 경 : MUG1은 고분자량의 당화 당단백으로 정상적으로 선상피의 선단부에서 발현되나, 종양세포에서는 비정상적인 발현 양상을 보인다. CD44는 세포-세포간 부착, 세포-기질간 부착에 관여하는 당단백이다. 일부 종양에서 MUC1 과 CD44는 종양의 침습과 전이에 관계한다고 알려져 있다. 본 연구에서는 비소세포폐암에서의 MUC1과 CD44의 standard form (CD44s)의 발현과 조직형, TNM-병기와의 상관성을 조사하였다. 대상 및 방법 : 수술적 방법으로 얻은 비소세포폐암 80예의 파라핀포매 조직절편을 대상으로 하여 MUC1과 CD44s에 대한 면역조직화학염색을 시행하였다. 결 과 : MUC1 은 편평상피암종의 12/43예 (27.9%), 선암종의 12/37예 (32.4%)에서 양성으로 조직형에 따른 차이는 보이지 않았다(p=0.660). CD44s는 편평상 피암종의 36/43예(83.7%), 선암종의 14/37예 (37.8%)에서 양성으로 선암종에 비해 편평상피암종에서 발현이 유의하게 높았다(p<0.001). 편평상피암종에서 TNM-병기에 따른 MUC1의 양성율은 통계적으로 유의한 차이를 보이지 않았다. 선암종에서 MUC1 양성은 N0에서 4/22예(18.2%), N1-2에서 8/15예 (53.3%)로 림프절 침범과 관련되었다(p=0.036). 편평상피암종에서 CD44s의 발현은 T-병기에 따른 의미있는 차이는 없었으나, N0기에서 16/21예 (72.7%), N1-2기에서 9/22예 (38.1%)로 CD44s의 발현의 감소는 림프절 침범과 관련되었다(p=0.031). 편평상피암종의 TNM-병기 I기에서 15/18예(83.3%), II-III기에서 10/25예 (40.0%)에서 CD44s 양성으로 CD44s 발현의 감소는 TNM-병기의 진행과 관련되었다(p=0.006). 결 론 : 비소세포폐암에서 MUC1과 CD44s는 종양의 조직형에 따라 다른 발현 양상을 보이고, 각각은 종양의 침습과 전이에 관여하는 것으로 보인다.

• IMMUNE NETWORK
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• 제11권6호
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• pp.368-375
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• 2011

Background: Recent clinical observation reported that there was a significant correlation between change in circulating vascular endothelial growth factor (VEGF) levels and the occurrence of severe acute graft-versus-host disease (GVHD) following allogeneic hematopoietic stem cell transplantation (allo-HSCT), but the action mechanisms of VEGF in GVHD have not been demonstrated. Methods: This study investigated whether or not blockade of VEGF has an effect on acute GVHD in a lethally irradiated murine allo-HSCT model of $B6\;(H-2^b)\;{\rightarrow}B6D2F1\;(H-2^{b/d})$. Syngeneic or allogeneic recipient mice were injected subcutaneously with anti-VEGF peptides, dRK6 ($50{\mu}g/dose$) or control diluent every other day for 2 weeks (total 7 doses). Results: Administration of the dRK6 peptide after allo-HSCT significantly reduced survival with greaterclinical GVHD scores and body weight loss. Allogeneic recipients injected with the dRK6 peptide exhibited significantly increased circulating levels of VEGF and expansion of donor $CD3^+$ T cells on day +7 compared to control treated animals. The donor $CD4^+$ and $CD8^+$ T-cell subsets have differential expansion caused by the dRK6 injection. The circulating VEGF levels were reduced on day +14 regardless of blockade of VEGF. Conclusion: Together these findings demonstrate that the allo-reactive responses after allo-HSCT are exaggerated by the blockade of VEGF. VEGF seems to be consumed during the progression of acute GVHD in this murine allo-HSCT model.

• 대한기관식도과학회지
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• 제3권1호
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• pp.70-78
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• 1997

The development of preneoplastic and neoplastic squamous cell proliferations of body sites such as the skin, female lower genital tract, and larynx is strongly associated with specific types of human papillomaviruses (HPV). Antitumor $CD^{8+}$ cells recognize peptide antigens presented on the surface of tumor cells by major histocompatibility complex (MHC) class I molecules. The MHC class I molecule is a heterodimer composed of an integral membrane glycoprotein designated the alpha chain and a noncovalently associated, soluble protein called beta-2-microglobulin( $\beta$ -2-m). Loss of $\beta$-2-m generally eliminates antigen recognition by antitumor $CD^{8+}$ T cells. We evaluated the expression of $\beta$-2-m as a potential means of tumor escape from immune recognition and the presence of HPV DNA as a cause of laryngeal squamous cell carcinomas (SCCs). Laryngeal SCCs (n=39) were analyzed for MHC class I expression by immunohistochemistry and for presence of HPV by in situ hybridization technique. The results were as follows : 1) HPV DNA was detected in 10 (25.64%) out of 39 cases in laryngeal squamous cell carcinomas. 2) MHC class I down-regulation (heterogenous and negative expression) in HPV positive lesions was higher than HPV negative lesions. 3) The expression of MHC class I was related to cellular differentiation regardless of T-stage and nodal involvement. In conclusion, HPV was thought to be the etiological factor of SCC of larynx, and we found that the down-regulation of MHC class I was a common phenomenon In laryngeal SCC and may provide a way for tumor cells to escape from immune surveillance.

• 대한본초학회지
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• 제32권5호
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• pp.27-38
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• 2017

Objective : In the present study, we examined whether Canavalia gladiata D.C. (CG) and Arctium lappa L., Redix (AL) mixture (CGAL), their components, lupeol and chicoric acid, regulate immune system and suppress the tumor in vitro and in vivo. Methods : LPS-induced reactive oxygen species (ROS) and nitric oxide (NO) were measured after treatment with CG extract (CGE), CGAL, lupeol, chicoric acid and lupeol and chicoric acid mixture (lupeol+CA) in Raw264.7 cell. To determine the effect of CGE on immune responses, immune cell population and IgG production were assessed in mice. To investigate the effect of CGAL and their component on anti-tumor activity, tumor volume and weight were measured, cell cycles and immune cell population were analyzed in MC38 injected tumor bearing mice. Also, NK cell activity was determined in splenocyte isolated from tumor bearing mice. Results : CGE, CGAL, lupeol, chicoric acid and lupeol+CA decreased the LPS-induced ROS and NO production without cell toxicity in RAW264.7 cells. CGE increased the immune cell populations of $CD4^+T$, $CD8^+T$ and macrophages in various immune organ of mice. In tumor bearing mice, CGAL, lupeol, chicoric acid and lupeol+CA suppressed tumor volume and weight. In cell cycle analysis, they decreased the percentages of S phase. In addition, CGAL, lupeol, chicoric acid and lupeol+CA immune cell populations of $CD4^+T$, $CD8^+Tcell$, NK cell and macrophage in tumor as well as NK cell activity. Conclusion : CGAL and its compounds may enhance immune responses and suppress tumor growth, and may be capable of developing health functional foods.

• BMB Reports
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• 제47권9호
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• pp.512-517
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• 2014

In this study, we showed that Mycobacterium abscessus MAB2560 induces the maturation of dendritic cells (DCs), which are representative antigen-presenting cells (APCs). M. abscessus MAB2560 stimulate the production of pro-inflammatory cytokines [interleukin (IL)-6, tumor necrosis factor (TNF)-${\alpha}$, IL-$1{\beta}$, and IL-12p70] and reduce the endocytic capacity and maturation of DCs. Using $TLR4^{-/-}$ DCs, we found that MAB2560 mediated DC maturation via Toll-like receptor 4 (TLR4). MAB2560 also activated the MAPK signaling pathway, which was essential for DC maturation. Furthermore, MAB2560-treated DCs induced the transformation of $na\ddot{i}ve$ T cells to polarized $CD4^+$ and $CD8^+$ T cells, which would be crucial for Th1 polarization of the immune response. Taken together, our results indicate that MAB2560 could potentially regulate the host immune response to M. abscessus and may have critical implications for the manipulation of DC functions for developing DC-based immunotherapy.

• Journal of Ginseng Research
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• 제46권5호
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• pp.690-699
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• 2022

Background: Few studies reported the therapeutic effect of Korean Red Ginseng (KRG) in lung inflammatory diseases. However, the anti-inflammatory role and underlying molecular in cadmium-induced lung injury have been poorly understood, directly linked to chronic lung diseases (CLDs): chronic obstructive pulmonary disease (COPD), cancer etc. Therefore, in this study we aim to investigate the therapeutic activities of water extract of KRG (KRG-WE) in mouse cadmium-induced lung injury model. Method: The anti-inflammatory roles and underlying mechanisms of KRG-WE were evaluated in vitro under cadmium-stimulated lung epithelial cells (A549) and HEK293T cell line and in vivo in cadmium-induced lung injury mouse model using semi-quantitative polymerase chain reaction (RT-PCR), quantitative real-time PCR (qPCR), luciferase assay, immunoblotting, and FACS. Results: KRG-WE strongly ameliorated the symptoms of CdSO4-induced lung injury in mice according to total cell number in bronchoalveolar lavage fluid (BALF) and severity scores as well as cytokine levels. KRG-WE significantly suppressed the upregulation of inflammatory signaling comprising mitogen-activated protein kinases (MAPK) and their upstream enzymes. In in vitro study, KRG-WE suppressed expression of interleukin (IL)-6, matrix metalloproteinase (MMP)-2, and IL-8 while promoting recovery in CdSO₄-treated A549 cells. Similarly, KRG-WE reduced phosphorylation of MAPK and c-Jun/c-Fos in cadmium-exposed A549 cells. Conclusion: KRG-WE was found to attenuate symptoms of cadmium-induced lung injury and reduce the expression of inflammatory genes by suppression of MAPK/AP-1-mediated pathway.