• Title/Summary/Keyword: mouse B cell

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Stimulatory Effects of Ginsan on the Proliferation and Viability of Mouse Spleen Cells

  • Ko, Eun-Ju;Joo, Hong-Gu
    • The Korean Journal of Physiology and Pharmacology
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    • 제14권3호
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    • pp.133-137
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    • 2010
  • Ginsan is an acidic polysaccharide purified from Panax ginseng, a famous oriental herb. Although a variety of biological activities of ginsan have been studied, the effects of ginsan on spleen cells are not fully elucidated. We investigated the effect of ginsan on the viability and proliferation of spleen cells. Using Cell Counting $Kit-8^{(R)}$ solution and trypan blue solution, we found that ginsan significantly enhanced viability and proliferation. Multiple clusters, indicating proliferation, were observed in ginsan-treated spleen cells and, carboxyfluorescein succinimidyl ester and surface marker staining assay revealed that ginsan promoted proliferation from $CD19^+$ B cells rather than $CD4^+$ or $CD8^+$ T cells. In addition, ginsan decreased the percentage of late apoptotic cells. Ginsan increased the surface expression of CD25 and CD69 as well as production of interleukin-2 from spleen cells, suggesting increased activation. Taken together, these results demonstrate that ginsan increases the viability and proliferation of spleen cells via multiple mechanisms, valuable information for broadening the use of ginsan in clinical and research settings.

Identification of Immunostimulatory Oligodeoxynucleotide from Escherichia coli Genomic DNA

  • Choi, Yong-Jun;Lee, Keun-Wook;Kwon, Hyung-Joo;Kim, Doo-Sik
    • BMB Reports
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    • 제39권6호
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    • pp.788-793
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    • 2006
  • Bacterial DNA containing immunostimulatory CpG motifs can stimulate antigen-presenting cells to express co-stimulatory molecules and to produce various cytokines in vivo and in vitro. In this study, we fragmented macromolecular E.coli genomic DNA with DNase I, and analyzed the ability of the resulting DNA fragments to induce the NF-${\kappa}B$ activation and humoral immune response. Furthermore, using computational analysis and luciferase assay for synthetic ODNs based on the sequence of the immunostimulatory DNA fragments (DF-ODNs), an active component of DF-ODNs sequences was investigated. Experimental results demonstrated that DF-ODN is optimal for the NF-${\kappa}B$-responsive promoter activation in the mouse macrophage cell line and the humoral immune response in vivo. In agreement with the activity of the DF-ODNs processed by DNase I, a synthetic ODN based on the DF-ODN sequences is potent at inducing IL-12 mRNA expression in primary dendritic cells. These results suggest that the discovery and characterization of a highly active natural CpG-ODN may be achieved by the analyses of bacterial DNA fragments generated by a nuclease activity.

수용성 누에 실샘 가수분해물의 미백 및 항노화 효과 (Whitening and Anti-aging Activities of Soluble Silkworm Gland Hydrolysate)

  • 정지혜;황정욱;김호진;차현명;김동일;최용수
    • KSBB Journal
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    • 제28권3호
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    • pp.196-201
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    • 2013
  • Silkworm-derived silk materials which contain 45 to 55% protein (18 amino acids, including 8 essential amino acids) have free radical-scavenging activities. In the present study, we investigated the whitening and anti-aging effects of silkworm gland under various conditions with B16F1 melanoma cells and human dermal fibroblasts. To evaluate the toxicity of soluble gland hydrolysate (SGH), mouse-derived B16F1 melanoma cells were treated with 0.2~5 mg/mL, cytotoxicity was not observed at all concentrations. The tyrosinase and elastase activities were significantly decreased as dosage levels of SGH increased. In addition, cell death was decreased by treated with SGH and antioxidative activity was the most effective in the SGH treatment. These findings suggest that SGH may be used as a potential functional biomaterial in having the skin whitening effect and anti-aging activity by inhibition of tyrosinase and elastase activities.

Identification of Genes Differentially Expressed in RAW264.7 Cells Infected by Salmonella typhimurium Using PCR Method

  • Kang, Kyung-Ho;Song, Jung-A;Shin, Dong-Jun;Choy, Hyon-E;Hong, Yeong-Jin
    • Journal of Microbiology
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    • 제45권1호
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    • pp.29-33
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    • 2007
  • Salmonella typhimurium, causing mouse typhoid, infects hosts such as macrophage cells, and proliferates in intracellular vacuoles causing infected cells to trigger numerous genes to respond against the infection. In this study, we tried to identify such genes in RAW264.7 cells by using the PCR screening method with degenerate primers. Fourteen genes were found to be differentially expressed after a 4 h infection in which the expression of 8 genes increased while expression of the others decreased. Most of the genes were involved in proinflammatory responses such as cytokines production and cell death. The mutation in msbB gene encoding the myristoyl transferase in lipid A of lipopolysaccharide (LPS) resulted in much lower toxicity to the inoculated animals. We compared the expression of the identified genes in wild-type and msbB-mutated S. typhimurium infections and found that Lyzs encoding lysozyme type M was differentially expressed. This gene is quite likely to be related to bacterial survival in the host cells.

도토리 추출물의 경구 투여가 마우스 면역 세포 활성에 미치는 효과 (Effects of Water Extract Acorn on Mouse Immune Cell Activation Ex Vivo)

  • 류혜숙
    • 한국식품영양학회지
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    • 제23권2호
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    • pp.135-140
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    • 2010
  • Acorns have been used as a traditional remed as well as food source. However, few studies on their immunomodulating effects have been reported. In this study, the combined immunomodulative effect of a water extract of acorns was tested on seven to eight weeks old mice(balb/c). The mice were fed ad libitum on a chow diet, and a water extract of the plant mixture was orally administered every other day for four weeks at two different concentrations(50 and 500 mg/kg B.W.). The production of cytokine(IL-$1{\beta}$, IL-6, IL-2, IL-10, IFN-$\gamma$), secreted by macrophages stimulated with LPS or not, detected by ELISA assay using cytokine kit. After 48 h of incubation with mitogen(ConA or LPS) ex vivo study showed that cytokine (IL-$1{\beta}$, IL-6, IL-2, IL-10, IFN-$\gamma$) was detected in both of the 50 and 500 mg/kg B.W. supplementation groups with LPS stimulation. The results of this study may suggest that supplementation with acorn water extract increase immune function by regulating cytokine production capacity by activated macrophages.

자두(후무사, 대석, 피자두) 추출물이 LPS로 염증을 유발한 Raw 264.7 세포와 암 세포에 미치는 영향 (Effect of Extracts from Oriental Plum (Formosa, Oishiwase, Soldam) on LPS-stimulated Raw 264.7 Cells)

  • 김세나;김소영;김정봉;박홍주;조영숙
    • 동아시아식생활학회지
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    • 제23권2호
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    • pp.197-202
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    • 2013
  • The objective of this study is to evaluate the anti-cancer and anti-inflammatory activities of plum (Formosa, Oishiwase, Soldam) for the future development of functional food products. To determine the anti-inflammatory effect of different types of plums, the inhibitory effect of plum extracts on nitric oxide (NO) production were measured in lipopolysaccharide (LPS)-stimulated Raw 264.7 mouse macrophage cells and human cancer cell lines (A549, Ags, Hela, Hep3B). Among the three different plum cultivars, Oishiwase at a concentration of 1 mg/mL showed the highest inhibitory effects on NO production (%) in Raw 264.7 macrophage cells. Moreover, Oishiwase exhibited a higher anti-cancer activity against A549 (renal carcinoma, 50%), Ags (gastric carcinoma, 35%), HeLa (cervical carcinoma, 50%), and Hep3B (hepatocellular carcinoma, 31%) at a concentration on 1 mg/mL, respectively, compared to Formosa and Soldam. Our findings suggest Oishiwase plum extracts may serve as potential dietary sources of natural health promoting substances.

덱스트로메트르판이 마우스의 일차 체액성 면역반응에 미치는 영향 (Effects of Dextromethorphan on the Primary Humoral Immune Response in Mice)

  • 표명윤
    • Environmental Analysis Health and Toxicology
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    • 제21권4호
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    • pp.349-355
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    • 2006
  • Dextromethorphan hydrobromide (DXM) has been widely used as a nonopioid antitussive drug with low toxicity and low potential for drug dependency. DXM is a psychotropic drug since 2003 in our country. This study was performed to investigate the immunotoxicity induced by abuse of DXM. Mice were orally exposed to DXM dissolved in saline as concentration of 30, 60, and 120 mg/kg b.w. before (day-2) or after (day+2) immunization (OVA-antigen, day 0). Thereafter, we measured the increased rate of body weight, relative weight of organ (thymus, spleen, liver, kidney) and OVA-specific IgM level in sera. In addition, mouse splenocytes were exposed to various concentration of DXM $(0.001{\sim}100{\mu}M)$ and cultured with B cell mitogen (LPS) and splenocytes proliferations (SP) were measured by MTT-assay. Thymus-weight were slightly changed on day 9 after administration of DXM, but body-, spleen-, liver-, and kidney-weight were not different between control group and DXM-treated group. SP to LPS were significantly decreased at high concentration $(100{\mu}M)$ when compared with controls. When DXM was administered before or after immunization with OVA-antigen, OVA-specific IgM levels were significantly lowered in a dose-dependent manner. These results indicate that DXM nay depress the primary humoral immune response to the initial antigenic challenge.

젖소 초유로부터 분리한 Insulin-like Growth Factor-1 분획이 In Vitro에서 마우스의 면역 활성에 미치는 영향 (Effects of IGF-I Rich Fraction from Bovine Colostral Whey on Immune Activity of Mouse In Vitro)

  • 황경아;양희진;이수원
    • 한국축산식품학회지
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    • 제26권1호
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    • pp.113-120
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    • 2006
  • 젖소 초유 whey 내에 존재하는 IGF-I을 분리하기 위하여 ultrafiltration을 사용하여 30과 1 kDa 사이의 IGF-I rich fraction을 분획하였다. 분획한 IGF-I rich fraction내에 IGF-I의 존재 여부는 SDS-PACE와 western blotting으로 확인하였고, sandwich ELISA로써 분획 내 IGF-I을 정량하였다. 그 결과 IGF-I은 단백질 mg당 10 ng으로 측정되었다. 분획한 IGF-I rich fraction을 마우스 복강 macrophge의 면역 활성에 미치는 영향 즉, IL-6, NO, $TNF-{\alpha}$, Phagocytosis, $H_{2}O_{2}$,의 분비 유도 및 생성량을 측정하였고, splenocyte의 면역 활성으로는 splenocyte의 증식 효과와 natural killer cell의 항암작용을 측정하였다. 실험 결과 IGF-I rich fraction $1{\mu}g/mL$ 투여군에서 IL-6의 생성량은 9.85 ng, NO의 생성량은 $17.1{\mu}M$, phagocytosis는 양성 대조구에 대비하여 78.3%의 탐식작용을 나타내었고 $TNF-{\alpha}$$H_{2}O_{2}$의 생성량은 대조구에 대비하여 각각 34.5 및 6% 더 많은 양을 생산하였다. Splenocyte 면역활성에 미치는 영향으로 T cell과 B cell 증식은 대조군에 대비하여 각각 36 및 76%로 더 높은 증식 효과를 나타내었고, NK cell의 활성은 대조군보다 55.4%의 높은 활성을 보였다.

LPS로 유발한 대식세포의 염증반응과 마우스 귀 부종에 대한 구멍갈파래 에탄올 추출물의 항염증 효과 (Anti-Inflammatory Effects of Ethanol Extracts from Ulva pertusa Kjellman on LPS-induced RAW 264.7 Cells and Mouse Model)

  • 김민지;김민주;김꽃봉우리;박선희;최현덕;박소영;장미란;임무혁;안동현
    • 한국미생물·생명공학회지
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    • 제44권4호
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    • pp.479-487
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    • 2016
  • 대식세포에 대하여 구멍갈파래 에탄올 추출물의 독성결과를 확인해본 결과 독성은 나타나지 않았으며, LPS에 의하여 유도되는 NO와 염증성 cytokine의 분비량은 구멍갈파래 에탄올 추출물의 농도 의존적으로 감소함을 확인하였다. 또한 구멍갈파래 에탄올 추출물로 인해 $NF-{\kappa}B$ 및 MAPKs의 신호전달을 억제함으로써 염증매개성 물질의 발현 억제에 효과가 있는지 알아본 결과, 구멍갈파래 에탄올 추출물은 각각 iNOS, COX-2, $NF-{\kappa}B$ 및 MAPKs의 활성을 효과적으로 억제하였고 그에 따른 염증 매개인자들의 생성도 효과적으로 억제되는 것을 확인하였다. 마지막으로 추출물이 마우스 귀부종에 미치는 영향을 살펴본 결과, 대조군의 경피와 진피의 두께에 비해 추출물 처리군의 조직 두께가 상대적으로 현저히 줄어들었으며 귀 조직에 침윤된 mast cell의 감소에도 추출물이 그 효과를 현저하게 나타냄을 확인하였다. 본 연구결과들을 종합해 보았을 때, 구멍갈파래의 에탄올 추출물은 항염증 활성을 가지는 새로운 천연물질로 이용 가능하여 고부가 가치 제품 개발이 가능한 천연 소재로 판단된다.

Listeria monocytogenes 신속 검출을 위한 면역크로마토그래피법의 개발 (Development of Immunochromatography for the Rapid Detection of Listeria monocytogenes)

  • 최진길;심원보;제정현;김지영;이규호;김민곤;하상도;김근성;김광엽;김철호;정덕화
    • 한국식품과학회지
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    • 제39권3호
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    • pp.299-303
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    • 2007
  • 본 연구에서는 L. monocytogenes를 신속하게 분석할 수 있는 면역크로마토그래피법을 개발하고자 하였다. 먼저 L. monocytogenes에 대한 단크론성 항체(FKLM-3B12-37)를 개발하여 직경 40nm로 제작된 colloidal gold와 합성한 후 conjugate pad에 처리하였고 nitrocellulose membrane 상의 test line과 control line에 단크론성 항체와 anti-mouse IgG를 각각 처리하여 면역크로마토그래피법을 개발하였다. 개발된 면역크로마토그래피법의 검출한계는 $10^{5}$ cell/mL 수준까지 검출이 가능했으며 다른 식중독 세균과는 교차 반응이 없는 것으로 나타났다. 임으로 오염시킨 식품시료의 경우 증균과정 없이 $10^{4}$ cell/25g까지 바로 분석하는데 어려움이 있으며 24시간의 증균과정을 거친 후에는 $10^{4}$ cell/25g까지 검출할 수 있었다. 본 연구에서 개발된 면역크로마토그래피법은 24시간의 증균 만으로 식품에 오염되어있는 L. monocytogenes를 검출할 수 있는 것으로 사료되었고, 기존의 식중독균 진단방법에 비해 신속하고 간편하게 그 결과를 확인할 수 있었다.