• Title/Summary/Keyword: Cell surface expression

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Anti-Oral Microbial Activity and Anti-Inflammatory Effects of Rosmarinic Acid in Lipopolysaccharide-Stimulated MC3T3-E1 Osteoblastic Cells on a Titanium Surface

  • Jeong, Moon-Jin;Lim, Do-Seon;Heo, Kyungwon;Jeong, Soon-Jeong
    • Journal of dental hygiene science
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    • v.20 no.4
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    • pp.221-229
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    • 2020
  • Background: The purpose of this study was to investigate the anti-oral microbial activity and anti-inflammatory effects of rosmarinic acid (RA) in lipopolysaccharide (LPS)-stimulated MC3T3-E1 osteoblastic cells on a titanium (Ti) surface during osseointegration, and to confirm the possibility of using RA as a safe natural substance for the control of peri-implantitis (PI) in Ti-based dental implants. Methods: A disk diffusion test was conducted to confirm the antimicrobial activity of RA against oral microorganisms. In order to confirm the anti-inflammatory effects of RA, inflammatory conditions were induced with 100 ng/ml of LPS in MC3T3-E1 osteoblastic cells on the Ti surface treated with or without 14 ㎍/ml of RA. The production of nitric oxide (NO) and prostaglandin E2 (PGE2) in LPS-stimulated MC3T3-E1 osteoblastic cells on the Ti surface was confirmed using an NO assay kit and PGE2 enzyme-linked immunosorbent assay kit. Reverse transcription polymerase chain reaction and western blot analysis were performed to confirm the expression of interleukin (IL)-1β and tumor necrosis factor (TNF)-α in total RNA and protein. Results: RA showed weak antimicrobial effects against Streptococcus mutans and Escherichia coli, but no antimicrobial activity against the bacteria Aggregatibacter actinomycetemcomitans and the fungus Candida albicans. RA reduced the production of pro-inflammatory mediators, NO and PGE2, and proinflammatory cytokines, TNF-α and IL-1β, in LPS-stimulated MC3T3-E1 osteoblastic cells on the Ti surface at the protein and mRNA levels. Conclusion: RA not only has anti-oral microbial activity, but also anti-inflammatory effects in LPS-stimulated MC3T3-E1 osteoblasts on the Ti surface, therefore, it can be used as a safe functional substance derived from plants for the prevention and control of PI for successful Ti-based implants.

Type I Collagen-induced Pro-MMP-2 Activation is Differentially Regulated by H-Ras and N-Ras in Human Breast Epithelial Cells

  • Kim, In-Young;Jeong, Seo-Jin;Kim, Eun-Sook;Kim, Seung-Hee;Moon, A-Ree
    • BMB Reports
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    • v.40 no.5
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    • pp.825-831
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    • 2007
  • Tumor cell invasion and metastasis are often associated with matrix metalloproteinases (MMPs), among which MMP-2 and MMP-9 are of central importance. We previously showed that H-Ras, but not N-Ras, induced invasion of MCF10A human breast epithelial cells in which the enhanced expression of MMP-2 was involved. MMP-2 is produced as a latent pro-MMP-2 (72 kDa) to be activated resulting the 62 kDa active MMP-2. The present study investigated if H-Ras and/or N-Ras induces pro-MMP-2 activation of MCF10A cells when cultured in two-dimensional gel of type I collagen. Type I collagen induced activation of pro-MMP-2 only in H-Ras MCF10A cells but not in N-Ras MCF10A cells. Induction of active MMP-2 by type I collagen was suppressed by blocking integrin ${\alpha}2$, indicating the involvement of integrin signaling in pro-MMP-2 activation. Membrane-type (MT)1-MMP and tissue inhibitor of metalloproteinase (TIMP)-2 were up-regulated by H-Ras but not by N-Ras in the type I collagen-coated gel, suggesting that H-Ras-specific up-regulation of MT1-MMP and TIMP-2 may lead to the activation of pro-MMP-2. Since acquisition of pro-MMP-2 activation can be associated with increased malignant progression, these results may help understanding the mechanisms for the cell surface matrix-degrading potential which will be crucial to the prognosis and therapy of breast cancer metastasis.

Changes of Immunoglobulins and Lymphocyte Subpopulations in Peripheral Blood from Holstein Calves Challenged with Escherichia coli Lipopolysaccharide

  • Kim, M.H.;Yun, C.H.;Kim, G.R.;Ko, J.Y.;Lee, Jung-Joo;Ha, Jong-K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.5
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    • pp.696-706
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    • 2011
  • The objective of this study was to characterize serum immunoglobulins and lymphocytes subpopulations in the peripheral blood mononuclear cells (PBMCs) of Holstein calves in response to lipopolysaccharide (LPS) challenge from Escherichia coli. Fourteen calves received subcutaneous injections of E. coli LPS at 10 weeks of age, and six calves were injected with saline as a control. The concentrations of total serum IgG and the relative amount of LPS-specific IgG in calves challenged with LPS were significantly higher (p<0.05) compared to control animals and LPS challenge significantly increased (p<0.05) the percentage of $CD5^+$ and $CD21^+$ T cells in PBMCs. Meanwhile, LPS challenge significantly increased (p<0.05, p<0.01) the percentage of $CD8^+$ and $CD25^+$ T cells in peripheral blood mononuclear cells (PBMC) at 7 and 14 Day-post LPS challenge (DPLC), respectively. The composition of $CD4^+CD25^+$ T cells and $CD8^+CD25^+$ T cells from calves challenged with LPS was also higher (p<0.05 and p = 0.562, respectively) than those of control calves at 14 DPLC. In conclusion, LPS challenge not only induces production of IgG with expression of B-cell immune response related cell surface molecules, but also stimulates activation of T-lymphocytes in PBMC. Our results suggest that LPS challenge in calves is a good model to elucidate cellular immune response against Gram-negative bacterial infections.

Significant Attenuation of Aden-associate Virus Gene Expression by Catechol-conjugated Heparin Surface Coating (카테콜기가 도입된 헤파린의 표면고정화에 의한 아데노연관바이러스의 발현 억제에 관한 연구)

  • Do, Minjae;Lee, Slgirim;Jang, Jae-Hyung;Lee, Haeshin
    • Journal of Adhesion and Interface
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    • v.17 no.4
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    • pp.149-154
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    • 2016
  • In this study, natural polymer-based virus neutralizing agent was developed in an attempt to replace the conventional sterilization method for mammalian cell culture. A catechol conjugated heparin was synthesized by using EDC chemistry, and it show unique binding ability to virus which has heparin affinity (adenovirus, adeno-associated virus). To evaluate neutralization ability of catechol conjugated heparin, adeno-associated virus was used for test model, instead of using a pathogenic virus. The catechol conjugated heparin exhibited resistance to high concentration of salt and complete inactivation of adeno-associated virus. The result suggests that the catechol conjugated heparin, which is biocompatible and efficiency, may replace conventional sterilization method for mammalian cell culture.

Cytotoxic T Lymphocytes Elicited by Dendritic Cell-Targeted Delivery of Human Papillomavirus Type-16 E6/E7 Fusion Gene Exert Lethal Effects on CaSki Cells

  • Wu, Xiang-Mei;Liu, Xing;Jiao, Qing-Fang;Fu, Shao-Yue;Bu, You-Quan;Song, Fang-Zhou;Yi, Fa-Ping
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.6
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    • pp.2447-2451
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    • 2014
  • Human papillomavirus (HPV) is the primary etiologic agent of cervical cancer. Consideration of safety and non human leukocyte antigen restriction, protein vaccine has become the most likely form of HPV therapeutic vaccine, although none have so far been reported as effective. Since tumor cells consistently express the two proteins E6 and E7, most therapeutic vaccines target one or both of them. In this study, we fabricated DC vaccines by transducing replication-defective recombinant adenoviruses expressing E6/E7 fusion gene of HPV-16, to investigate the lethal effects of specific cytotoxic T lymphocytes (CTL) against CaSki cells in vitro. Mouse immature dendritic cells (DC) were generated from bone marrow, and transfected with pAd-E6/E7 to prepare a DC vaccine and to induce specific CTL. The surface expression of CD40, CD68, MHC II and CD11c was assessed by flow cytometry (FCM), and the lethal effects of CTL against CaSki cells were determined by DAPI, FCM and CCK-8 methods. Immature mouse DC was successfully transfected by pAd-E6/E7 in vitro, and the transfecting efficiency was 40%-50%. A DC vaccine was successfully prepared and was used to induce specific CTL. Experimental results showed that the percentage of apoptosis and killing rate of CaSki cells were significantly increased by coculturing with the specific CTL (p <0.05). These results illustrated that a DC vaccine modified by HPV-16 E6/E7 gene can induce apoptosis of CaSki cells by inducing CTL, which may be used as a new strategy for biological treatment of cervical cancer.

Chemotactic Effect of Leukotactin-1/CCL15 on Human Neutrophils

  • Lee Ji-Sook;Yang Eun-Ju;Ryang Yong-Suk;Kim In-Sik
    • Biomedical Science Letters
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    • v.12 no.3
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    • pp.145-151
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    • 2006
  • Leukotactin-l (Lkn-l )/CCL15 has been known as a potent chemoattractant of leukocytes. However, the precise function of Lkn-l in human neutrophils has not been explained well. In the present study, we investigated the contribution of Lkn-1 in chemotactic activity of human neutrophils. Both CCR1 and CCR3 mRNA expressions are strongly expressed in human neutrophils but CCR2 protein expression was uniquely detected on the cell surface. Lkn-l binding to CCR1 and CCR3 induced chemotactic activity of neutrophils. Chemotactic index of Lkn-l was comparable to that of IL-8. $MIP-1{\alpha}/CCL3$ binding to CCR1 and CCR5 has no effect on neutrophil migration. Cell migration, in response to Lkn-l, was blocked by pertussis toxin (Ptx), a $G_o/G_i$ protein inhibitor, and U73122, a phospholipase C(PLC) inhibitor but not by protein kinase C inhibitor such as rottlerin, and Ro-31-8425. Taken together, our results demonstrate that Lkn-l transduces the chemotaxis signal through $G_o/G_i$ protein and PLC. This finding provides the molecular mechanism by which Lkn-l may contribute to neutrophil movement into the site of inflammation.

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Study on the Anti-inflammatory Effects of Soyangin-Hyeongbangpaedok-san (소양인 형방패독산의 함염증효과에 대한 실험적 연구)

  • Heo, Jeong-Won;Kang, Hee;Ahn, Kwang-Seok;Kim, Sung-Hoon;Choi, Seung-Hoon;Ahn, Kyoo-Seok;Shim, Bum-Sang
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.2
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    • pp.443-451
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    • 2009
  • Soyangin-Hyeongbangpaedok-san(SHBPDS) is used for treating upper respiratory infections, In an effort to investigate the anti-inflammatory effects of SHBPDS, we measured production of several cytokines and immunoglobulin in various immune cells. SHBPDS decreased the secretion of TNF-${\alpha}$, but not that of IL-6 in PMA/A23187 stimulated HMC-1 cells. As for mouse B cells, it induced proliferation and caused differential effects in expressions of surface IgE as determined by flow cytometry and secretions of IgE, IgG1, ILA and INF-${\gamma}$as measured by ELISA but showed little change in CD23 or CD69 expression. SHBPDS increased proliferation in anti-CD3/anti-CD28 stimulated CD4 Th cells. Under the Th1/Th2 polarization conditions, SHBPDS at 200 ${\mu}g/m{\ell}$ suppressed the secretion of INF-${\gamma}$ and IL-4. Based on the above results, we conclude that SHBPDS has antiinflammatory activities in mast cells and different immunomodulatroy effects in B cells and Th cells.

Effects of Auklandia Lappa on Dextran Sulfate Sodium-Induced Murine Colitis (당목향(唐木香)이 DSS(Dextran sulfate sodium)로 유발된 염증성 장질환 동물모델에 미치는 영향)

  • Kim, So-Yeon;Park, Jae-Woo;Ryu, Bong-Ha
    • The Journal of Internal Korean Medicine
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    • v.34 no.2
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    • pp.134-146
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    • 2013
  • Objectives : Auklandia Lappa (ALE) is one of the herbs used frequently to treat abdominal pain and diarrhea and reported anti-inflammatory activity by suppressing proinflammatory cytokines. This study was designed to investigate whether ALE could show protective activities on experimental colitis induced by dextran sulfate sodium (DSS) models. Methods : Colitis was induced by DSS in Balb/c mice. ALE 10, 30, 100 and 300 mg/kg were orally administered twice a day for 7 days in DSS model. Mice weight was measured daily. Scoring of clinical findings was measured every other day. Colon length, edema, fecal blood and histological damages were assessed at day 7 in DSS model. In histological analysis, we checked cryptal glands, surface epithelium, submucosa, transmural, stroma and scored degree of inflammatory cell damage by modified histological scoring. We also calculated cytokines concentrations including IFN-${\gamma}$, TNF-${\alpha}$, IL-6, IL-$1{\beta}$, IL-17, IL-23, IL-10 and TGF-${\beta}1$ by Biometric Multiplex Cytokine Profiling method. Results : ALE showed the protective effects on DSS-induced experimental colitis. ALE inhibited shortening of colon length and histological damages of colon does-dependently, but it did not inhibit weight loss. ALE also inhibited IFN-${\gamma}$ and IL-6 expression, and upregulated cytokines (IL-10, TGF-${\beta}1$) related to regulatory T cell differentiation and proliferation. Conclusions : The current results demonstrate the clinical utility of ALE in traditional medicine and indicate the possibility of potent drug development of inflammatory bowel diseases from natural products. Further investigations for exact mechanisms will be needed.

Circulating Aneuploid Cells Detected in the Blood of Patients with Infectious Lung Diseases

  • Kim, Hongsun;Cho, Jong Ho;Sonn, Chung-Hee;Kim, Jae-Won;Choi, Yul;Lee, Jinseon;Kim, Jhingook
    • Journal of Chest Surgery
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    • v.50 no.2
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    • pp.126-129
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    • 2017
  • The identification of circulating tumor cells (CTCs) is clinically important for diagnosing cancer. We have previously developed a size-based filtration platform followed by epithelial cell adhesion molecule immunofluorescence staining for detecting CTCs. To characterize CTCs independently of cell surface protein expression, we incorporated a chromosomal fluorescence in situ hybridization (FISH) assay to detect abnormal copy numbers of chromosomes in cells collected from peripheral blood samples by the size-based filtration platform. Aneuploid cells were detected in the peripheral blood of patients with lung cancer. Unexpectedly, aneuploid cells were also detected in the control group, which consisted of peripheral blood samples from patients with benign lung diseases, such as empyema necessitatis and non-tuberculous mycobacterial lung disease. These findings suggest that chromosomal abnormalities are observed not only in tumor cells, but also in benign infectious diseases. Thus, our findings present new considerations and bring into light the possibility of false positives when using FISH for cancer diagnosis.

Size, Shape, and Crystal Structure-dependent Toxicity of Major Metal Oxide Particles Generated as Byproducts in Semiconductor Fabrication Facility (반도체 가공 작업환경에서 부산물로 발생되는 주요 금속산화물의 입자 크기, 형상, 결정구조에 따른 독성 고찰)

  • Choi, Kwang-Min
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.26 no.2
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    • pp.119-138
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    • 2016
  • Objectives: The purpose of this study is to review size, shape, and crystal structure-dependent toxicity of major metal oxide particles such as silicon dioxide, tungsten trioxide, aluminum oxide, and titanium dioxide as byproducts generated in semiconductor fabrication facility. Methods: To review the toxicity of major metal oxide particles, we used various reported research and review papers. The papers were searched by using websites such as Google Scholar and PubMed. Keyword search terms included '$SiO_2$(or $WO_3$ or $Al_2O_3$ or $TiO_2$) toxicity', 'health effects $SiO_2$(or $WO_3$ or $Al_2O_3$ or $TiO_2$). Additional papers were identified in references cited in the searched papers. Results: In various cell lines and organs of human and animals, cytotoxicity, genotoxicity, hepatoxicity, fetotoxicity, neurotoxicity, and histopathological changes were induced by silicon dioxide, tungsten trioxide, aluminium oxide, and titanium dioxide particles. Differences in toxicity were dependent on the cell lines, organs, doses, as well as the chemical composition, size, surface area, shape, and crystal structure of the particles. However, the doses used in the reported papers were higher than the possible exposure level in general work environment. Oxidative stress induced by the metal oxide particles plays a significant role in the expression of toxicity. Conclusions: The results cannot guarantee human toxicity of the metal oxide particles, because there is still a lack of available information about health effects on humans. In addition, toxicological studies under the exposure conditions in the actual work environment are needed.