Hericium erinaceum induces maturation of dendritic cells derived from human peripheral blood monocytes

Hericium erinaceum, a medicinal mushroom, has long been used as a therapeutic due to its immuno‐regulating potentials eliciting anticarcinogenic and antimicrobial efficacies. Since maturation of dendritic cells (DC) is an important process in the initiation and regulation of immune responses, the ability of water‐soluble components from H. erinaceum (WEHE) to regulate DC maturation was investigated. Immature DC were prepared by differentiating human peripheral blood CD14‐positive cells with GM‐CSF and IL‐4. DC were stimulated with WEHE at 2–20 µg/mL for 48 h and subjected to flow cytometric analysis to determine the expression of indicative maturation markers. The endocytic capacity of WEHE‐stimulated DC was examined by a Dextran‐FITC uptake assay. An enzyme‐linked immunosorbent assay was performed to examine the secretion of TNF‐α and IL‐12p40. DC stimulated with WEHE showed representative features upon DC maturation: enhanced expression of CD80, CD83 and CD86, and both MHC class I and II molecules, decreased endocytic capacity of DC, increased expression of CD205, and decreased expression of CD206. However, interestingly, WEHE could not induce the production of TNF‐α and IL‐12p40, whereas lipopolysaccharide substantially increased the production of both cytokines. Collectively, these results suggest that H. erinaceum induces the maturation of human DC, which might reinforce the host innate immune system. Copyright © 2009 John Wiley & Sons, Ltd.     

Effects of bee venom on the maturation of murine dendritic cells stimulated by LPS

Aim of study

This study was performed to elicit the effectiveness of bee venom (BV), a traditional immunosuppressive Korean acupuncture agent, on the maturation of dendrtic cells (DCs).

Materials and methods

Immature dendritic cells (iDCs) were generated from mouse bone marrow cells with GM-CSF. After 10 days of initial differentiation, DCs were activated with lipopolysaccharides (LPS) for another 48h in the presence or absence of BV. Surface molecule analysis, intracytoplasmic staining of cytokines, FITC-conjugated antigen uptake, and transwell migration assays were conducted with iDCs and activated DCs.

Results

Up-regulation of costimulatory molecules, typical of mature DCs (mDCs) was inhibited by addition of BV. Pro-inflammatory cytokines were also found to be reduced with BV treatment in LPS-stimulated DC. A decrease in antigen uptake upon the maturation of DC was reversed in low dose BV treated mDC. In addition, BV treated mDC demonstrated reduced directional migration in response to CCL21, a lymphoid chemokine which directs mDC.

Conclusions

BV may have a therapeutic effect an on abnormally activated immune status, such as autoimmune rheumatoid arthritis, through an immune-modulatory effect on DC.     

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??OBJECTIVE To study the active part of Achyranthes bidentata promotes maturation and regulates the functions of DC in tumor microenvironment. METHODS The total saponins and the polysaccharides from dry powder of Achyranthes bidentata were extracted and identified. The tumor microenvironment by co-culturing breast cancer MCF-7 cells with DCs, and add polysaccharides, total saponins or total saponins with polysaccharides of Achyranthes bidentata to stimulate DCs were established. The expression of surface molecules on DCs and the killing activity of cytotoxic T lymphocytes (CTL) induced by DCs by Flow Cytometry were detected. ELISA was used to detect the levels of IL-1??, TNF-??, IL-10 and IL-12 in the culture supernatant. Detect the protein expression of JAK, p-JAK, STAT and p-STAT3 in DCs by Western blot. RESULTS In the tumor microenvironment, the active parts of Achyranthes bidentata, total saponins and ABPS, alone or combined used, can trigger JAK/STAT3 signaling pathway of DCs through TLR4 and Dectin-1 receptor and upregulate the expression of surface molecules, such as CD80, CD86 and CD40, to induce the maturation and differentiation of DCs, promote the secretion of TNF-??, IL-12, IL-10 and IL-1??. CONCLUSION As an effective adjuvant, the active parts of traditional Chinese medicine can enhance the effect of DC vaccine to correct the immune deficiency in tumor microenvironment, activate tumor-specific CTLs and then produce specific anti-tumor effects.     

大黄素体外抑制人树突状细胞分化及成熟作用的研究

目的 探讨大黄素体外作用树突状细胞（dendritic cell, DC）后对人DC分化、成熟及免疫功能的影响。     

Toll-like receptor ligand-induced activation of murine DC2.4 cells is attenuated by Panax notoginseng

The medicinal herb, Panax notoginseng, has been used for thousands of years in traditional Chinese medicine and possesses anti-inflammatory properties. Dendritic cells (DCs) play a central role in the regulation of both inflammation and adaptive immunity. The aim of this study was to investigate the potential for notoginseng extracts to modulate Toll-like receptor (TLR) ligand-induced activation of cultured DC2.4 cells. Following stimulation with LPS, CpG or poly(I:C) and treatment with 0-50micorg/ml notoginseng extract for 24 h, DCs were evaluated for various phenotypic and functional readouts. Notoginseng reduced the LPS-, CpG- and poly(I:C)-induced production of TNF-alpha by DC2.4 cells. Also, IL-6 production by notoginseng-treated cells stimulated with LPS and CpG but not poly(I:C) was reduced when compared to controls. TLR ligand-induced CD40 expression was attenuated by notoginseng. In contrast, notoginseng decreased CD86 levels on DCs activated with LPS and poly(I:C) but not CpG. Inhibition of TNF-alpha production was time-dependent in LPS-stimulated cells, occurring only with pretreatment or concurrent treatment of notoginseng but not after delayed addition of the herbal extract. Additionally, ginsenoside Rg1 more effectively inhibited LPS-stimulated cytokine production by DC2.4 cells than ginsenoside Rb1. Taken together, these results demonstrate that notoginseng inhibits the production of specific inflammatory molecules and innate immune responsiveness by DCs following TLR activation.     

黄芪提取物免疫调节活性的体外实验研究

目的：研究黄芪提取物（Astragalus membranaceus extract,AME)对人的外周血免疫细胞（PBIC）免疫功能的调节作用。方法：采用^3H-TdR掺入法分析AME对外周血单个核细胞（PBMC）的增殖活性及对外周血粘附单核细胞（PBAM）吞噬肿瘤细胞的影响；采用^51Cr释放法测定AME对杀伤性T细胞（CTL）杀伤肿瘤细胞的影响；用ELISA法和生物学法研究AME对外周血B细胞（PBBC）产生IgG及对PBAM产生细胞因子的影响；采用SDS－PAGE分析AME的蛋白组成。结果：AME能促进PBMC的增殖；提高CTL对肿瘤细胞的杀伤活性；增强PBAM对肿瘤细胞的吞噬和产生细胞因子的功能；促进了PBBC产生IgG;AME含有多种蛋白成分。结论：AME对人免疫功能有增强作用，提高了抗肿瘤免疫效应，可应用于临床调节免疫功能和治疗肿瘤等疾病。     

Aqueous extracts from Menyanthes trifoliate and Achillea millefolium affect maturation of human dendritic cells and their activation of allogeneic CD4+ T cells in vitro

Ethnopharmacological relevance: Menyanthes trifoliate and Achillea millefolium have been used in traditional medicine to ameliorate chronic inflammatory conditions. The aim of this study was to identify the effects of ethanol and aqueous extracts of Menyanthes trifoliate and Achillea millefolium on maturation of dendritic cells (DCs) and their ability to activate allogeneic CD4⁺ T cells.

Materials and methods

Human monocyte-derived DCs were matured in the absence or presence of lyophilised aqoueous or ethanol extracts from Menyanthes trifoliate or Achillea millefolium and their expression of surface molecules analysed with flow cytometry and cytokine secretion measured by ELISA. DCs matured in the presence of aqueous extracts from Menyanthes trifoliate and Achillea millefolium were co-cultured with allogeneic CD4⁺ T cells and the expression of surface molecules by T cells and their cytokine secretion and cell proliferation determined.

Results

Maturation of DCs in the presence of aqueous extracts from Menyanthes trifoliate or Achillea millefolium did not affect expression of the surface molecules examined but reduced the ratio of secreted IL-12p40/IL-10, compared with that by DCs matured in the absence of extracts. Allogeneic CD4⁺ T cells co-cultured with DCs matured in the presence of aqueous extract from Menyanthes trifoliate secreted less IFN-γ, IL-10 and IL-17 than CD4⁺ T cells co-cultured with DCs matured without an extract. Maturation of DCs in the presence of aqueous extract from Achillea millefolium decreased IL-17 secretion but did not affect IFN-γ and IL-10 secretion by allogeneic CD4⁺ T cells.

Conclusions

Aqueous extract from Menyanthes trifoliate induces a suppressive phenotype of DCs that has reduced capacity to induce Th1 and Th17 stimulation of allogeneic CD4⁺ T cells, whereas aqueous extract from Achillea millefolium reduces the capacity of DCs to induce a Th17 response.     

人参总皂苷对DC2.4细胞增殖、抗原吞噬和表面分子表达的影响

目的:观察人参总皂苷对树突状细胞系DC2.4细胞增殖、抗原吞噬和表面分子表达的影响。方法:体外培养DC2.4细胞,加入不同剂量人参总皂苷后,MTT法检测不同时点细胞增殖;流式细胞仪检测细胞表面分子CD40、CD86和MHCⅡ的表达,以及对卵清蛋白抗原的吞噬作用。结果:单纯加入脂多糖(LPS)经培养48h后DC2.4细胞增殖明显(P0.05),人参总皂苷可明显抑制LPS诱导的DC2.4细胞增殖(P0.05);此外,人参总皂苷可促进DC2.4细胞吞噬抗原的能力,并协同LPS上调CD40、CD86和MHCⅡ分子表达。结论:人参总皂苷可以维持DC2.4细胞数量的稳态,促进DC2.4细胞吞噬抗原,并与LPS协同促进DC2.4细胞成熟,并具有促进抗原提呈的作用。     

Enhancement of Extracts from Celastrus orbiculatus on Maturation and Function of Dendritic Cells in vitro and in vivo

Objective To examine the immunoregulation of Celastrus orbiculatus extracts (COE), a traditional Chinese medicine, on maturation and function of dendritic cells (DCs) in vitro and in vivo. Methods In vitro, after treated with COE in different nontoxic concentrations (0, 10, 20, 40, 80, and 160 μg/mL) for 5 d, the surface immunological molecules and cytokine secretion of mice bone marrow-derived DCs in response to COE were analyzed by flow cytometric analysis (FACS) and enzyme linked immunosorbent assay (ELI...     

Panax quinquefolium saponins inhibited immune maturation of human monocyte-derived dendritic cells via blocking nuclear factor-κB pathway

Ethnopharmacological relevance

Panax quinquefolium saponins (PQS), a water-soluble antioxidant extracted from a natural herb, radix panacis quinquefolii (American Ginseng), has yielded encouraging results in the treatment of atherosclerotic diseases. However, the underlying mechanisms remain unclear. Here, we tested the hypothesis that the anti-atherosclerotic effect of PQS might be mediated by suppressing human monocyte-derived dendritic cells (DCs) maturation.

Materials and methods

DCs were derived by incubating purified human monocytes with granulocyte macrophage colony stimulating factor (GM-CSF) and IL-4. DCs were pre-incubated with or without PQS and stimulated by oxidized low density lipoprotein (ox-LDL). Expression of DCs membrane molecules (CD40, CD86, CD1a, HLA-DR) and endocytotic ability were analyzed by FACS, cytokines (IL-12 and TNF-α) were measured by ELISA. Nuclear factor (NF)-κB signaling pathway was determined by Western blotting, and RT-PCR. NF-κB activation was quantified by ELISA.

Results

PQS reduced ox-LDL induced immunophenotypic expressions (CD40, CD1a, CD86, and HLA-DR) and cytokine secretions (IL-12 and TNF-α), and improved endocytotic ability of DCs. These above phenomena were accompanied by decreased protein expression and binding activity of nuclear localized c-Rel subunit.

Conclusions

Our study suggested that PQS inhibited ox-LDL induced immune maturation of DCs in vitro, which might be in part mediated by NF-κB signal transduction pathway.     

不同树种树枝对蜜环菌生长的影响

选用北方常见树种的树枝,接种培养蜜环菌,比较蜜环菌的生长速度、胞外漆酶酶活力及菌索的生长状况.结果显示:除了槐树、紫穗槐外,蜜环菌在桃树、梨树等多种树枝上均能正常生长,有希望用于栽培天麻的菌枝、菌棒.     

黄芪多糖诱导成熟的树突状细胞肿瘤疫苗体外抗肿瘤作用的实验研究

目的探讨经黄芪多糖（astragalus polysacharin,APS）诱导成熟的树突状细胞（dendritic cell,DC）肿瘤疫苗体外抗肿瘤作用及其机制。方法分离人外周血单个核细胞（peripheral blood mononuclear cell,PBMC）,制备DC细胞,加入营养液体外诱导为未成熟的DC细胞,培养第5天,黄芪多糖组加入终浓度为100μg/mL的黄芪多糖,细胞因子组加入终浓度为20 ng/mL重组人肿瘤坏死因子（rhTNF）-α诱导DC成熟,观察树突状细胞形态及表型变化;成熟DC以SGC-7901肿瘤抗原致敏,与同种异体T细胞共培养,采用MTT法检测T淋巴细胞增殖功能,E LISA法检测共培养上清IL-12、IFN-γ水平;经DC激活的细胞毒性T淋巴细胞（cytotoxic lymphocyte,CTL）细胞与SGC-7901肿瘤细胞共培养,采用LDH释放法检测CLT对靶细胞特异性杀伤能力。结果黄芪多糖诱导的DC经形态学观察和表型鉴定,符合成熟DC的特征。黄芪多糖诱导成熟的DC能刺激同种异体T淋巴细胞增殖,T细胞增殖指数随刺激细胞与效应细胞比值的增加而增加（P〈0.05）;DC与T细胞共培养上清IL-12、IFN-γ的水平显著增加且呈时间依赖性（P〈0.05）;经致敏DC激活的CTL能显著杀伤肿瘤细胞,随着效靶比的增加,杀伤作用增强。结论黄芪多糖可在体外诱导DC成熟,并促进其抗原递呈能力,有效活化CTL,增强机体抗肿瘤功能。     

Immunomodulatory effects of aqueous and organic fractions from Petiveria alliacea on human dendritic cells

Petiveria alliacea is a plant traditionally known for its anti-inflammatory and anti-tumor activities; however, the molecular and cellular mechanisms of its immunomodulatory properties are still unknown. Dendritic cells (DC) promote adaptive immune response by activating T lymphocytes, inducing an effector response or tolerance depending on the DC differentiation level. Herein, we evaluated the immunomodulatory activity of aqueous and organic plant fractions from P. alliacea using human monocyte-derived dendritic cells. The phenotype, cytokine secretion and gene expression were estimated after treatment with the plant fractions. We found that P. alliacea aqueous fraction induced morphological changes and co-stimulatory expression of CD86, indicating partial DC maturation. In addition, pro-inflammatory cytokines such as IL-1β, IL-6, IL-8, IL-10, IL-12p70, and TNF-α were secreted. The fraction also increased NF-κB gene expression while down-regulating TGFβ gene expression. These results suggest that the aqueous fraction can induce partial DC activation, a situation that can be relevant in tolerance induction. It is important to state that the organic fraction by itself does not show any immunomodulatory activity. This study provides evidence for possible immunomodulatory activity of P. alliacea extracts which has been used in traditional medicine in Colombia.     

人参皂苷Rb1对氧化型低密度脂蛋白诱导的人单核细胞源树突状细胞免疫成熟的影响

目的 研究中药单体人参皂苷Rb1(ginsenoside Rb1, GRb1)对氧化型低密度脂蛋白(oxidized low density lipoprotein, OX-LDL)诱导的人单核细胞源树突状细胞(dendritic cells, DCs)免疫成熟的影响。     

不同浓度丁酸钠诱导未成熟树突状细胞分化的免疫学功能比较

 目的 探讨不同浓度丁酸钠诱导对于体外培养的树突状细胞（dendritic cells, DC）免疫刺激活性的影响。方法 通过重组人粒细胞-巨噬细胞集落刺激因子(GM-CSF)和人重组白细胞介素4（IL-4）体外诱导的人单个核细胞来源的DC,按照0.5、0.75、1、1.25、1.5 mmol·L-15种浓度丁酸钠体外诱导分化,并分别以流式细胞仪,FITC-dxtran内吞检测、MLR、ELISA法检测DC的凋亡率、表面标志、内吞能力、刺激淋巴细胞增殖能力和IL-10、IL-12、INF-γ分泌的变化。结果 1.25、1.5 mmol·L-1丁酸钠有较高的细胞毒性。0.5、0.75、1 mmol·L-1丁酸钠均可抑制DC表面成熟表型的表达。其中以1.0 mmol·L-1丁酸钠诱导下DC不成熟表型表达效果最稳定。虽然不同浓度丁酸钠诱导下的DC内吞能力无显著差异（P>0.05）,但1.0 mmol·L-1丁酸钠与0.5、0.75 mmol·L-1丁酸钠相比较其IL-10分泌量（3.29±0.21）最大,抑制淋巴细胞增殖能力（1.53±1.04）最强（P<0.05）。结论 1.0 mmol·L-1丁酸钠是诱导DC稳定不成熟状态的最佳浓度。     

加味青蒿鳖甲汤对髓系MRD-L患者CD_(34)~+细胞源DC诱导过程中IL-12的影响

目的:探讨加味青蒿鳖甲汤对树突细胞(DC)诱导和对IL-12分泌的影响。方法:本研究采用体外培养的方法,用含不同剂量(高、中、低)加味青蒿鳖甲汤动物血清联合细胞因子、不含药血清联合细胞因子及单纯用细胞因子,共同培养缓解期急性髓系白血病(AML-CR)患者骨髓中分离提取的CD+34细胞,诱导其成为DC,观察各组在不同阶段对DC成熟的影响,以及各组血清中IL-12的分泌变化。结果:含药血清均能上调DC表面特征性分子及共刺激分子CD80、CD83、CD86表达水平,较单纯细胞因子组比较有显著性差异(P<0.01),且含中剂量药血清更能促进DC分泌IL-12(P<0.01)。结论:联合细胞因子的含加味青蒿鳖甲汤的血清,可使来源AML-CR骨髓的CD+34,在体外比单纯用细胞因子诱导培养DC能够更好地促进DC的生长、成熟和分化,并能促进DC分泌IL-12,增加抗肿瘤作用。     

环孢素A对小鼠髓源性树突状细胞的影响

 目的研究环孢素A(CsA)对小鼠髓源性树突状细胞(BMDC)的影响。方法以小鼠BMDC为对象,在其培养过程中加入不同质量浓度的CsA(0,0.5和1 mg·L^-1)、脂多糖(LPS)诱导细胞成熟,流式细胞分析其免疫表型的改变,氚-胸腺嘧啶核苷(³H-TdR)掺入法和ELISA法检测其刺激同种T细胞增殖能力和分泌细胞因子等功能变化。结果CsA处理后的树突状细胞(DC),其表面共刺激分子CD80,CD86表达下调,抑制性分子B7-DC表达进一步上调,而另一抑制性分子B7-H1表达无明显变化,该DC刺激同种T细胞增殖能力下降,TNF-α和IL-12P70分泌降低,IL-10分泌增加。结论在BMDC的分化过程中,CsA能直接作用于DC,使其具有不成熟DC的特性,该DC可以诱导免疫耐受,这种作用是通过共刺激分子CD80,CD86表达下调和抑制性分子B7-DC表达上调实现的。     

华蟾素对慢性乙型肝炎患者外周血来源树突状细胞的影响

目的:研究华蟾素对慢性乙肝患者外周血来源的树突状细胞(dendritic cells,DCs)形态、表型和功能的影响,以了解华蟾素提高慢乙肝患者机体免疫力的作用机制。方法:实验研究分3组:正常对照组、慢乙肝组和慢乙肝华蟾素组。分离慢性乙肝患者和正常人外周血单核细胞(Monocytes,Mo),以GM-CSF和IL-4联合诱导培养生成DCs,慢乙肝华蟾素组于DCs培养48小时后加入华蟾素。各组DCs均于培养的第7天以相差显微镜观察和拍摄细胞形态;用流式细胞仪检测表面分子CD40、HLA-DR、CD80、CD86的表达水平;用CCK8法检测DCs刺激同种异体淋巴细胞增殖的能力;用ELISA法检测DCs培养液上清中IL-12的水平。结果:与正常人相比,慢乙肝组患者外周血来源的DCs细胞形态相对不成熟,表面分子表达水平低,刺激同种异体淋巴细胞增殖能力和IL-12分泌能力较弱;加入华蟾素培养的慢乙肝患者DCs与慢乙肝组比较,细胞形态相对成熟,表面分子CD40、CD80和CD86表达水平增高,刺激同种异体淋巴细胞增殖能力和IL-12分泌能力增强。结论:华蟾素能促进慢性乙肝患者外周血来源DCs的发育成熟,改善其功能。