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

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® 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.     

Targeting Vaccines to Dendritic Cells

Dendritic cells (DC) are specialized antigen presenting cells (APC) with a remarkable ability to take up antigens and stimulate major histocompatibility complex (MHC)-restricted specific immune responses. Recent discoveries have shown that their role in initiating primary immune responses seems to be far superior to that of B-cells and macrophages. DC are localized at strategic places in the body at sites used by pathogens to enter the organism, and are thereby in an optimal position to capture antigens. In general, vaccination strategies try to mimic the invasiveness of the pathogens. DC are considered to play a central role for the provocation of primary immune responses by vaccination. A rational way of improving the potency and safety of new and already existing vaccines could therefore be to direct vaccines specifically to DC. There is a need for developing multifunctional vaccine drug delivery systems (DDS) with adjuvant effect that target DC directly and induce optimal immune responses. This paper will review the current knowledge of DC physiology as well as the progress in the field of novel vaccination strategies that directly or indirectly aim at targeting DC.     

阿奇霉素对支气管哮喘患儿外周血树突状细胞功能的影响

目的:观察阿奇霉素对支气管哮喘患儿外周血单个核细胞(PBMC)来源树突状细胞(DCs)功能的影响,探讨阿奇霉素免疫调节作用的机制.方法:选择16例支气管哮喘发作期患儿为研究对象,18例健康儿童为对照组.无菌条件下采用密度梯度离心法获取PBMC,体外经rhGM-CSF、rhIL-4诱生DCs.哮喘组置于不同浓度阿奇霉素(...     

Effect of Low Dose Radiation on Differentiation of Bone Marrow Cells into Dendritic Cells

Low dose radiation has been shown to be beneficial to living organisms using several biological systems, including immune and hematopoietic systems. Chronic low dose radiation was shown to stimulate immune systems, resulting in controlling the proliferation of cancer cells, maintain immune balance and induce hematopoietic hormesis. Since dendritic cells are differentiated from bone marrow cells and are key players in maintaining the balance between immune activation and tolerance, it may be important to further characterize whether low dose radiation can influence the capacity of bone marrow cells to differentiate into dendritic cells. We have shown that bone marrow cells from low dose-irradiated (γ-radiation, 0.2Gy, 15.44mGy/h) mice can differentiate into dendritic cells that have several different characteristics, such as expression of surface molecules, cytokine secretion and antigen uptake capacity, when compared to dentritic cells differentiated from the control bone marrow cells. These differences observed in the low dose radiation group can be beneficial to living organisms either by activation of immune responses to foreign antigens or tumors, or maintenance of self-tolerance. To the best of our knowledge, this is the first report showing that total-body low dose radiation can modulate the capacity of bone marrow cells to differentiate into dendritic cells.     

Immunomodulatory effects of Ziziphora tenuior L. extract on the dendritic cells

Background

Ziziphora tenuior L. (Kakuti in Persian) is used in traditional medicine for treatment of gastrointestinal disorders as carminative and analgesic plant. The other usages of this plant are included treatment of diarrhea and nausea. Therefore in the present study we evaluated the immunomodulatory effects of the ethanolic extract of this plant on the dendritic cells (DCs).

Results

Ziziphora tenuior L. extract significantly (p = 0.002) increased the level of surface expression of CD40 as an important co-stimulatory marker on DCs compared to the control. However this extract did not change CD86 and MHC-II molecules, so it could promote DCs phenotypic maturation. Treatment of DCs with the extract resulted in slightly increased of the production of (IL-12); however, this change was not significant. In addition, the ability of treated DCs to stimulate allogenic T cells proliferation and cytokines secretion was examined in the co-cuture of these cells with T cells in mixed lymphocyte reaction (MLR). Z. tenuior L. at the 100 μg/ml concentration inhibited the proliferation of allogenic T cells and also significantly (P < 0.001) increased the level of IL-10. Moreover, the extract at 10–100 μg/ml concentration caused slightly increase in IFN-γ production and decreased IL-4 cytokines but these changes were not significant.

Conclusions

These findings indicated that Z. tenuior L. extract can modulate immune response by induction of CD40 expression on DCs and cytokine production; whereas it can inhibit T cell stimulating activity of DCs in high concentration. These findings possibly in part explain the traditional use of this plant in treatment of immune-mediated disorders. However future studies are needed.     

hKDR融合基因致敏树突状细胞对小鼠肝癌细胞杀伤活性研究

目的:探讨hKDR融合基因致敏树突状细胞(DC)激发的细胞毒性T淋巴细胞(CTL)对小鼠肝癌细胞的杀伤活性.方法:自小鼠骨髓中获取DC,以粒细胞-巨噬细胞集落刺激因子(GM-CSF)和hKDR融合基因的mRNA致敏DC;用致敏的DC免疫小鼠取其脾脏获得CTL,即效应细胞,用脂质体将pCDNA hKDR转染至小鼠肝癌Hepal-6细胞株中,作为靶细胞;将效应细胞与靶细胞按不同比例混合,用乳酸脱氢酶(LDH)释放法检测细胞毒作用.结果:未经修饰的DC激活的CTL与靶细胞比例为100:1、50:1、25:1情况下,其杀伤率分别为19.2%、12.3%、6.9%;经hKDR修饰的DC激活的CTL与靶细胞比例为100:1、50:1、25:1情况下,其杀伤率分别为71.6%、55.8%、22.7%,将两组结果在相同的比例下两两比较,均有显著性差异(P＜0.05).结论:hKDR融合基因致敏DC激活的CTL对小鼠肝癌细胞具有较强的杀伤作用,且杀伤能力随效应细胞与靶细胞比例增高而增强.     

Immunomodulatory Activity of n-Butanol Extract of Oroxylum indicum

The present study evaluated the immunomodulatory activity and mechanism of action of the n-butanol extract [100 mg/kg body weight (b.w.), per oral (p.o.)] of Oroxylum indicum Vent. (Bignoniaceae) root bark against different experimental animal models, i.e., immune response to sheep red blood cells in stress-induced immunosuppression, carbon clearance assay and neutrophil adhesion test. In the immune response to sheep red blood cell model, the n-butanol extract of Oroxylum indicum treatment group showed a significant rise (P < 0.05) in circulating antibody-titer as compared to the stress-induced control group. This observation suggests that the drug treatment caused potentiation of humoral antibody immune response to antigen. Also, it significantly decreases the rise in malondialdehyde content along with an increase (p < 0.05) in superoxide dismutase, catalase and reduced glutathione levels as compared to the stress-induced control group and, thus, exhibits significant antioxidant potential. Pretreatment with the n-butanol extract of Oroxylum indicum showed a significant rise (P < 0.05) in phagocytic index indicating phagocytic activity. In the neutrophil adhesion model, a significant increase (P < 0.05) in percentage neutrophil adhesion was observed in the n-butanol extract treatment group as compared to the control group. From these findings, the n-butanol extract of Oroxylum indicum possesses immunomodulatory activity by enhancing specific immune response (humoral immunity) and non-specific immune response (phagocytosis) of the body as well as exhibiting antioxidant potential. Our results suggest that the n-butanol extract of Oroxylum indicum root bark possesses a significant immunostimulant activity.     

虫草菌多糖对吸烟引起的树突状细胞氧化损伤的保护作用

目的:研究虫草菌多糖(EPS)对树突状细胞系(DCS)因吸烟引起的氧化损伤的保护性作用方法:采用噻唑蓝(MTT)法检测香烟提取物(CSE)对DCS生长的抑制作用;采用流式细胞技术检测CSE对DCS细胞产生活性氧(ROS)的影响以及在加入EPS保护的情况下CSE对DCS细胞产生ROS的影响;采用比色法检测有EPS保护的情况下,CSE对DCS细胞产生乳酸脱氢酶(LDH)的水平、过氧化氢酶(CAT)和超氧化物歧化酶(SOD)活力的影响.结果:CSE对DCS的生长有抑制性作用,且能够提高ROS的产生;在加入EPS后,EPS能够抑制ROS的产生以及减少细胞中LDH的量,提高SOD和CAT的活力.结论:EPS对DCS的氧化损伤起到了保护性作用.     

冻融抗原冲击致敏的树突状细胞抗肿瘤作用的研究

目的研究MC-38肿瘤冻融抗原致敏的树突状细胞(DC)对荷瘤小鼠是否有抗肿瘤作用。方法用MC-38肿瘤冻融抗原体外冲击致敏小鼠骨髓来源的DC,观察其诱导的CTL对MC-38肿瘤细胞的杀伤活性;体内以1×106DC/只多次接种于已荷瘤小鼠同侧腹股沟皮下,观察抗原冲击的DC对肿瘤生长的抑制作用以及对荷瘤小鼠生存期的影响。结果体外抗原冲击致敏的DC诱导的CTL对MC-38肿瘤细胞具有显著的杀伤作用,在效靶比为50∶1,25∶1,12.5∶1,6.25∶1时其杀伤率分别为68.84%,58.36%,41.56%,24.96%,;抗原冲击致敏的DC体内多次皮下免疫后对肿瘤的生长具有显著的抑制作用,能显著延长荷瘤小鼠的生存期。结论肿瘤冻融抗原致敏DC多次皮下免疫对荷瘤小鼠具有显著的抗肿瘤作用。     

苦瓜多糖抗肿瘤及免疫增强活性的研究

目的：研究苦瓜多糖的体内抗肿瘤及增强免疫作用,探讨可能作用机制。方法：通过建立小鼠S180肉瘤、H22肝癌肿瘤模型观察苦瓜多糖的抗肿瘤作用．此外应用MTT法观察苦瓜多糖对小鼠脾淋巴细胞增殖作用的影响．应用比色法观察苦瓜多糖对小鼠单核巨噬细胞RAW264．7吞噬功能的影响．运用Gfiess试剂检测苦瓜多糖促进小鼠单核巨噬细胞RAW264．7分泌No含量的变化。结果：苦瓜多糖高中低剂量组能明显抑制小鼠S180肉瘤、H22肝癌肿瘤的生长,同时能明显增加荷瘤小鼠的脾腺指数和胸腺指数。此外苦瓜多糖能显著刺激小鼠脾淋巴细胞增殖．明显提高小鼠单核巨噬细胞RAW264．7吞噬中性红的能力,明显促进小鼠单核巨噬细胞RAW264．7分泌No。结论：推测苦瓜多糖具有较强的免疫增强活性并可能通过刺激淋巴细胞、并增强巨噬细胞的活化来调节机体的免疫功能．从而抑制肿瘤细胞的生长。     

脐血来源树突状细胞增强CIK细胞对肺癌细胞株的杀伤作用

目的探讨肺癌细胞株A549肿瘤冻融抗原负荷脐血来源树突状细胞(DCs)与CIK细胞混合培养后对A549细胞杀伤活性的影响作用。方法取对数生长期的A549细胞制备肿瘤抗原(Ag),用脐血单个核细胞(CBMC)分别制备DCs和CIK细胞;用负荷Ag的DCs和CIK细胞共培养,诱导Ag-DC-CIK细胞。用流式细胞术检测DC免疫表型,MTT法检测CBMC、CIK、Ag-CIK和Ag-DC-CIK对A549肿瘤细胞株的杀伤活性。结果脐血DCs高表达CD86、CD40,中度表达CD83和CD80,低表达CD11c。实验组细胞(Ag-DC-CIK)对A549细胞的杀伤能力(62%)明显高于对照组(P<0.01),结论肿瘤抗原负荷的DCs能增强CIK细胞对靶细胞的杀伤活性,为DCs的免疫治疗提供了新的思路。     

The effects of cichorium intybus extract on the maturation and activity of dendritic cells

Background

Cichorium intybus is a medicinal plant commonly used in traditional medicine for its benefits in immune-madiated disorders. There are several evidences showing that C. intybus can modulate immune responses. In the present study we have investigated the effects of the ethanolic root extract of this plant on the immune system by targeting dendritic cells (DCs). For this purpose, phenotypic and functional maturity of murine DCs after treatment with the extract was analyzed by flow cytometry and mixed lymphocyte reaction (MLR) assay.

Results

C. intybus did not change the expression of CD40, CD86 and MHC-II molecules as important co-stimulatory markers on DCs compared to the control, indicating that it could not promote DCs phenotypic maturation. Treatment of DCs with lower concentrations of the extract resulted in an increased production of IL-12 by these cells with no change in IL-10 release. The capacity of treated DCs to stimulate allogenic T cells proliferation and cytokines secretion was examined in the co-cuture of these cells with T cells in MLR. C. intybus at higher concentrations inhibited proliferation of allogenic T cells and in lower concentrations changed the level of cytokines such that IL-4 decreased and IFN-γ increased.

Conclusions

These results indicated that C. intybus extract at higher concentrations can inhibit T cell stimulating activity of DCs, whereas at lower concentrations can modulate cytokine secretion toward a Th1 pattern. These data may in part explain the traditional use of this plant in treatment of immune-mediated disorders.     

异喹啉类生物碱的免疫调节作用*

异喹啉类生物碱是自然界存在的种类最多的一大类生物碱,具有抗肿瘤、抗炎、免疫调节和镇痛等多种生物活性。本文综述了阿朴菲类、双苄基异喹啉类、小檗碱类、吗啡类和苯菲啶类异喹啉生物碱对机体免疫应答的调节作用和机制,为其深入研究与开发提供参考。     

异喹啉类生物碱的免疫调节作用*

异喹啉类生物碱是自然界存在的种类最多的一大类生物碱,具有抗肿瘤、抗炎、免疫调节和镇痛等多种生物活性。本文综述了阿朴菲类、双苄基异喹啉类、小檗碱类、吗啡类和苯菲啶类异喹啉生物碱对机体免疫应答的调节作用和机制,为其深入研究与开发提供参考。     

人参多糖的研究概况

人参为五加科Araliaceae植物Panax ginseng C.A.Mey.的干燥根,在我国有着悠久的药用历史,主要含有人参皂苷、人参多糖、人参多肽及挥发油、多种氨基酸、脂肪酸及维生素、微量元素等有效成分,国内外对人参的活性成分的研究始终非常活跃,特别是对于人参皂苷的报道较多见,但是近年来随着分析手段的逐步提高,国内外学者对于人参多糖的研究也取得了较大的进展.     

三相静态萃取高效液相色谱法鉴别人参、三七及西洋参

目的建立人参、三七和西洋参的高效液相色谱（HPLC）鉴别方法，对其进行鉴别和质量控制。方法利用一种新的样品处理方法，即液-液-液三相静态萃取法对药材样品进行预处理，然后对三相萃取物分别以不同的流动相梯度进行反相高效液相色谱（RP-HPLC）法测定，记录色谱图和三维图谱。结果3种药材的HPLC图均具有良好的分离度和重现性，鉴别特征差异显著。结论三相静态萃取能够对参类药材的复杂成分进行有效的分离。药材各相萃取物的HPLC图均具有良好的鉴别特征，其中中间相较轻相和重相萃取物的色谱信息量大、专属性强，因此可以将中间相的特征色谱作为人参、三七和西洋参鉴别及质量指标量化的依据。     

Effects of polysaccharide ginsan from<Emphasis Type="Italic">Panax ginseng</Emphasis> on liver function

Ginsan, a polysaccharide isolated from Panax ginseng, has been shown to be a potent immunomodulator, producing a variety of cytokines such as TNF-alpha, IL-1, IL-2, IL-6, IL-12, IFN-gamma and GM-CSF, and stimulating lymphoid cells to proliferate. In the present study, we analyzed some immune functions 1st-5th days after ginsan i.p. injection, including the level of non-protein thiols (NPSH) as antioxidants, heme oxygenase (HO) activity as a marker of oxidative stress, zoxazolamine-induced paralysis time and level of hepatic cytochrome P-450 (CYP450) as indices of drug metabolism system, and activities of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), total bilirubin, and albumin level as indicators of hepatotoxicity. Ginsan in the dose of 100 mg/kg caused marked elevation (1.7 to approximately 2 fold) of HO activity, decrease of total CYP450 level (by 20-34%), and prolongation of zoxazolamine-induced paralysis time (by 65-70%), and showed some differences between male and female mice. Ginsan treatment did not seem to cause hepatic injury, since serum AST, ALT, and ALP activities and levels of total bilirubin and albumin were not changed.     

吡咯烷二硫代氨基甲酸酯对树突状细胞成熟的影响

目的：观察NF-κB抑制剂吡咯烷二硫代氨基甲酸酯（PDTC）对树突状细胞（DC）成熟的影响。方法：利用PDTC作用于体外培养的大鼠不成熟DC，细菌脂多糖（LPS）刺激后，流式细胞仪检测DC共刺激分子CD80、CD86、CD40表达；ELISA分析DC分泌IL—12的变化，NF—κB DNA结合ELISA检测核内NF—κB活性。结果：PDTC抑制DC共刺激分子CD80、CD86、CD40表达；减少DC分泌IL-12；抑制核内NF—κB DNA结合活性。结论：PDTC抑制DC成熟。     

银杏叶提取物对树突状细胞和Th1/Th2细胞因子的影响

目的研究银杏叶提取物对不稳定型心绞痛患者(UAP)树突状细胞(DC)功能和Th1/Th2细胞因子的影响。方法将54例UAP分为常规治疗组(对照组,27例)和常规加银杏叶提取物组(银杏叶组,27例),分别于治疗前及治疗后4周取血,分离外周血单个核细胞,在含粒细胞、巨噬细胞、集落刺激因子和白介素4的培养条件下制备DC。用流式细胞仪检测DC表面CD86(B7-2)的表达;混合淋巴细胞反应(MLR)检测DC对同种异体T淋巴细胞的刺激能力;ELISA法测定血液中的细胞因子IFN-γ和IL-4;比浊法测定血C反应蛋白(CRP)。体外实验观察银杏内酯B对DC CD86表达的直接影响。结果与对照组比较,银杏叶提取物可进一步抑制DC功能(P<0.01),同时显著降低血CRP水平(P<0.05)和血IFN-γ浓度(P<0.01);银杏内酯B对DC CD86表达有直接的抑制作用(P<0.01)。结论银杏叶提取物对DC的抑制和由此对患者炎性反应的抑制可能是其治疗心绞痛的机制之一。