人外周血单核细胞体外诱导成熟和激活的树突状细胞

目的　建立从人外周血单核细胞体外诱导培养成熟和激活的树突状细胞 (dendritic cell,DC)的方法。方法　从健康成人外周血分离单核细胞 (PBM) ,加入粒单系集落刺激因子 (GM-CSF) 10 0 μg/L+重组白细胞介素 -4 (rh IL -4 ) 5 0 0 k U /L体外培养 14 d,并于培养结束前 1d加入或不加入肿瘤坏死因子α (TNF -α ) (10 0μg/L ) ,流式细胞仪测定树突状细胞 (DC)的主要组织相容性复合体 (MHC) - 类分子、粘附分子和协同刺激分子 ,分析其成熟度和激活度。结果　 PBM经 GM-CSF+ rh IL-4诱导培养 14 d后 ,细胞成簇 ,表型为 CD83 2 2 .6%、CD865 5 .5 %、CD11c 3 6.1%、CD64 3 .2 %、人类白细胞抗原 (HLA) -DR 13 .4% ;培养结束前 1d加入 TNF -α诱导后 ,细胞表型为 CD83 81.5 %、CD8699.3 %、CD11c 98.8%、CD64 3 .4%、HLA-DR 88.3 %。结论　 GM-CSF +rh IL-4诱导 PB-M14 d,可获得大量不成熟的 DC,该体系有利于 DC扩增 ;培养结束前 1d加入 TNF-α,DC成熟度高 ,激活性好 ,适合于肿瘤免疫治疗     

西罗莫司对小鼠树突状细胞CD（86）、I-Ab和Toll样受体4表达的影响

目的：研究西罗莫司（SRL）对小鼠骨髓源树突状细胞（DC）发育成熟和Toll样受体4（TLR4）mRNA表达的影响。方法：（1）用细胞因子定向诱导C57BL/6小鼠骨髓细胞分化为DC,在相差显微镜和扫描电镜下动态观察DC在分化发育不同阶段的典型形态。（2）用SRL处理DC,通过流式细胞仪测定CD11c、CD86、MHC-Ⅱ类分子（I-Ab）的表达及在脂多糖（LPS）刺激后各分子表达的变化。（3）实时定量PCR法检测SRL处理DCTLR4mRNA表达水平。结果：（1）在相差显微镜和扫描电镜下可以看到DC在分化发育不同阶段的典型形态。（2）SRL抑制LPS刺激后DC表面CD86和I-Ab表达的上调。（3）与常规培养的DC相比,SRL组DCTLR4mRNA表达水平明显增高。结论：SRL处理的DC可抵抗LPS的促成熟作用。SRL促进DCTLR4mRNA表达。     

肺癌抑制免疫功能的靶细胞：树突状细胞

目的：了解肿瘤微环境下肺癌细胞对DC前体细胞产生DC、功能的影响。方法：DC前体细胞为健康人外周血CD14细胞，加入GM－CSF和IL－4，37℃，5％CO2培养7－10天收获，肿瘤细胞株CRL－5815，CRL－5826加入DC前体细胞共培养，健康入外周血为正常对照，FITC或PE交链的单克隆抗体标记DC细胞，流式细胞仪分析检测CD14，CD86，CD80，CD1a阳性表达。Annexin V方法，流式细胞仪分析细胞凋亡，混合淋巴细胞反应（MLR）检测DC检测T细胞反应的能力。结果：肺癌细胞株可诱导DC形成的不同时期产生凋亡，对DC产生的早期（第1、2天）影响较晚期（第6，7天）更明显，肺癌细胞株CRL－5815和CRL－5826与DC共培养时明显抑制DC细胞表型CD80，CD86，CD40的表达，与肺癌细胞株共培养DC其刺激T细胞扩增的能力（MLR）显下降，结论：肺癌细胞株CRL－5815和CRL－5826与DC前体细胞共培养时可导致DC的凋亡显增加，同时DC细胞表面表型表达下降，刺激T细胞扩增的功能减低，提示肿瘤（肺癌细胞）可通过DC细胞数量减少和功能下降影响机体免疫功能。     

体外培养成人脂肪间充质干细胞生物学特性及诱导分化为心肌细胞的研究

目的通过分离、培养脂肪间充质干细胞观察其生物学特性及诱导分化为心肌细胞,为心肌再生提供良好的干细胞来源。方法胶原酶消化分离成人脂肪来源的间充质干细胞并进行传代培养,倒置相差显微镜观察细胞形态,流式细胞仪测定CD29、CD31、CD34、CD44及细胞周期,MTT绘制细胞生长曲线。用第3代细胞进行诱导分化,观察不同浓度5-氮杂胞苷(5-Aza,1,3,5,10,15,20μmol/L)及不同作用时间(12,24,48,72h)诱导其向心肌细胞分化的差别,采用最佳浓度10μmol/L,最佳作用时间24h进行实验,分别在第7,14,21,28天用免疫细胞荧光染色鉴定心肌细胞α-横纹肌、肌球蛋白重链(MHC)、心肌肌钙蛋白I(cTnI)表达,第14天反转录-聚合酶链反应(RT-PCR)法检测心肌发育相关基因NKX2.5的表达。结果倒置相差显微镜下观察原代细胞,可见细胞呈梭型、核圆形或椭圆形,偶见双核。传代细胞核原代细胞形态相似,排列有了一定的方向性。流式细胞仪检测结果显示,第1、3、5代细胞均高表达CD29和CD44;而CD31始终表达很弱,可认为呈阴性表达;CD34在第1、3代细胞弱表达,在第5代细胞表达逐渐减弱为阴性。细胞生长曲线显示前3d处于细胞潜伏状态,第4天进入对数生长期,第10天达到顶峰。细胞周期检测结果显示G1期细胞为85.93%,S期为7.24%,G2期为6.83%。10μmol/L5-Aza诱导后7d进行免疫细胞荧光染色,未见有α-横纹肌、MHC、cTnI表达。14d少量细胞α-横纹肌和MHC阳性表达,cTnI阴性表达。21d表达α-横纹肌和MHC的细胞数量增多,并可见少量cTnI阳性表达。28dα-横纹肌、MHC、cTnT阳性表达数目均增多,RT-PCR结果显示NKX2.5呈阳性表达。结论成人脂肪中可以分离出脂肪间充质干细胞并且可以在体外培养传代,经过5-Aza的诱导可以向心肌细胞分化,为干细胞移植治疗和组织工程学种子细胞提供了更多的选择。     

人脂肪间充质干细胞分离培养及其生物学特性的实验研究

目的探索人脂肪间充质干细胞（adipose tissue—derived mesenchymal stem cells，ADMSCs）分离培养的方法及体外扩增的条件，观察ADMSCs的生物学特性。方法以腹部手术患者皮下脂肪组织为材料，采用I型胶原酶消化法及贴壁法分离培养ADMSCs，在含10％胎牛血清的低糖DMEM培养基中贴壁培养，倒置显微镜观察，流式细胞仪检测细胞表面标记CD29、CD44、CD105、CD31、CD34、CD106的表达，透射电镜及扫描电镜下观察ADMSCs超微结构，流式细胞仪测定细胞周期。结果原代和传代细胞呈梭形外观，生长增殖能力良好。CD29、CD44、CD105均呈阳性表达，阳性率分别为95.3％、98．6％和86．5％；而CD31、CD34、CD106阳性率分别为3．5％、2．6％、1．3％。透射电镜观察显示ADMSCs表现出早期幼稚细胞形态的特点，流式细胞仪检测显示84．8％的细胞处于G0/G1期。结论酶消化法能有效地从人脂肪组织分离培养人ADSCs，细胞生长稳定，增殖能力活跃，为今后ADMSCs的分离培养提供了更简单有效的方法。     

Fas配体基因逆转录病毒途径转染骨髓CD_(34)~+细胞

目的　借助逆转录病毒途径 ,FasL基因转染骨髓CD34+3 4 细胞。方法　将人FasLcDNA顺向插入逆转录病毒载体质粒PLXSN ;PLXSN -hFasL质粒脂质体转染PA317包装细胞 ;G418筛选抗生克隆 ;PCR鉴定整合细胞 ;扩增 ;制毒 :病毒滴度测定。试剂盒分选骨髓CD34+3 4 细胞 ;流式细胞仪分析其纯度 ;体外扩增培养 (IL -3+IL -6 +SCF) ;病毒上清转染CD34+3 4 细胞 ;流式细胞仪检测FasL阳性细胞率。结果　成功构建PLXSN -hFasL质粒 ;PA317细胞进行包装后所获假性病毒上清滴度为 1.7× 10 5CFU/ml ;对 11份正常人或良性贫血患者髓血进行CD34+3 4 细胞分选 ,所得CD34+3 4 细胞数为 1.35± 0 .45× 10 5,纯度为 85 %～ 97% ;体外扩增培养 10～ 12d ,细胞数达到 2 .5± 0 .7× 10 6(扩增约 18.5倍 ) ;病毒上清转染后 48～ 72h ,流式细胞仪检测 2 4.2± 2 .4%CD34+3 4 细胞表达FasL(P <0 .0 1) (未转染的CD34+3 4 细胞FasL阳性率为 7.6± 1.1% )。结论　通过逆转录病毒途径 ,FasL基因可有效转染骨髓CD34+3 4 细胞     

大黄素对体外诱导人血来源树突状细胞的影响

目的探讨大黄素对体外诱导培养的人血来源树突细胞(DC)的影响。方法分离健康人外周血单核细胞,经培养后获得未成熟DC(iDC),重组人粒-巨噬细胞集落刺激因子(rhGM-CSF,106IU.L-1)和重组人白细胞介素4(rhIL-4,8×105IU.L-1)诱导获得成熟DC(mDC)。实验分为iDC组、mDC组和大黄素组,mDC组于d 5加入脂多糖(LPS,1 mg.L-1)刺激,大黄素组采用mDC在d 5经LPS刺激后于d 7加入大黄素(100 mg.L-1)共培养2 d。用倒置显微镜和电镜观察DC细胞形态,用流式细胞仪检测DC表面共刺激分子的表达水平。结果经rhGM-CSF、rhIL-4诱导和LPS刺激,培养9 d后可获得表面有丰富分叉状胞浆突起的毛刺状mDC。大黄素组DC表面突起短而少,呈iDC形态。大黄素组CD80、CD83和CD86表达率分别为(13.4±6.6)%、(9.3±2.2)%和(84.2±6.3)%,低于mDC组[分别为(39.3±8.6)%、(30.7±5.6)%和(95.4±3.2)%,P<0.01];大黄素组CD14表达率为(8.4±2.8)%显著高于mDC组的(3.7±2.3)%(P<0.01)。大黄素组HLA-DR及CD11c表达与mDC组比较无显著差异(P>0.05)。结论大黄素能干扰DC表面突起的形成和表面共刺激分子的表达。     

脐血来源树突状细胞的培养与鉴定

目的建立并鉴定脐血来源树突状细胞(DC)培养体系，在体外诱导脐血干细胞生成成熟DC。探讨其在抗肿瘤免疫中的作用。方法脐血CD34 来源的DC以两步法，即0天开始使用GM-CSF、TNF-α、SCF、TPO、FL20，8～14天用GM—CSF、TNF—α和IL-4诱导生成。并分别在形态(光学显微镜 电镜)、表面标记(流式细胞仪)和功能(同种混合淋巴细胞反应)上加以鉴定。结果在合适的细胞因子组合下，能够经过脐血干细胞培养得到成熟的DC，具备典型的DC形态并在体外有效刺激同种淋巴细胞增殖。结论通过合适的培养可以获得大量成熟D(：。     

鸡尾酒法体外培养人外周血来源的树突状细胞

目的 探讨从健康人外周血中体外诱导培养成熟树突状细胞(DC)的方法.方法 用密度梯度离心法分离人外周血单个核细胞(PBMC),再用贴壁法获取单核细胞后加入人粒细胞-巨噬细胞集落刺激因子(GM-CSF)和IL-4,培养6 d后分组:Ⅰ组为对照组,仅含上述细胞因子;Ⅱ组加入多聚次黄嘌呤胞嘧啶核苷酸(Poly I∶C);Ⅲ组加入钙离子载体(CI)A23187;Ⅳ组加入Poly I∶C和CI A23187.第8天收获细胞,显微镜下观察形态,流式细胞术检测DC表型,体外同种混合淋巴细胞反应检测DC刺激T细胞的增殖活性.结果 Ⅳ组细胞形态学观察可见典型DC特征,荧光激活细胞分离器(FACS)检测高表达CD83、CD80、CD86和CD40;混合淋巴细胞反应(MLR)表明Ⅳ组细胞具有较强的刺激T细胞增殖能力.结论 PBMC体外经过细胞因子序贯、联合诱导培养能够获得大量功能成熟的DC.     

成人骨髓基质细胞体外培养及其生物学特性

目的：探索体外培养脑出血后遗症患者骨髓基质细胞(BMSCs)的方法及扩增后的生物学特性。方法取脑出血后3个月以上患者髂骨骨髓组织,去除淋巴细胞后采用静置贴壁培养法接种培养,经多次换液纯化。在倒置相差显微镜下观察细胞形态,取第3、6代应用流式细胞仪检测细胞表面抗原CD29、CD34、CD45、CD105因子的表达。结果倒置相差显微镜下接种48 h后,发现少数细胞贴壁,呈梭形,6~8 d后细胞形成集落。传代后,细胞形态趋于一致,可成漩涡状。流式细胞仪检测显示CD34、CD45阴性,而CD29、CD105阳性。结论体外密度梯度离心法和贴壁静置可以获得大量高度均一的表型符合骨髓基质细胞的细胞。     

Phenotypic and functional maturation of murine dendritic cells (DCs) induced by purified Glycyrrhizin (GL)

The aim of this study is to investigate phenotypic and functional modulation of murine dendritic cells (DCs) with use of purified Glycyrrhizin (GL). These impacts of GL on DCs both from bone marrow derived DCs and established DC cell 2.4 were assessed with conventional scanning electron microscopy (SEM), flow cytometry (FCM), transmission electron microscopy (TEM), cytochemistry assay, FITC-dextran, bio-assay and enzyme linked immunosorbent assay (ELISA). We found that the purified GL induced phenotypic maturation as evidenced by increased expression of CD86, CD40, CD80, CD83 and major histocompatibility complex II (MHC II). The functional tests showed the activity of acidic phosphatase (ACP) inside the DCs2.4 cells were down- regulated after treatment with GL (which occurs when phagocytosis of DCs2.4 cells were decreased). Finally, we proved that GL increased the production of IL-12, IL-10 and decreased the production of tumor necrosis factor alpha (TNF-α). These data indicated that GL could promote maturation of DCs and this adjuvant-like activity may have potential therapeutic value. It is therefore concluded that GL could exert positive modulation on murine DCs.     

Effects of vasoactive intestinal peptide on phenotypic and functional maturation of dendritic cells

The present study was designed to investigate the effects of vasoactive intestinal peptide (VIP) on differentiation, maturation of dendritic cells (DCs) in vitro. DCs were derived from the murine bone marrow hemopoietic progenitor cells by culturing in RPMI 1640 complete medium supplemented with GM-CSF and IL-4 in the presence or absence of various concentrations of vasoactive intestinal peptide (VIP) and lipopolysaccharide (LPS). The phenotype of DCs was analyzed by flow cytometry. Mixed leukocyte reaction (MLR) was employed to measure the capacity of DC to stimulate the allogeneic T cells. IL-12p70 secretion by DC was examined by ELISA. In the absence of LPS, VIP, in a dose dependent manner, up-regulated the expression of CD80, CD86, CD54 and CD40, but down-regulated the expression of MHC class II molecule (Ia(b)). In the presence of LPS, VIP also dose dependently up-regulated the expression of CD80, CD86, CD54 and CD40, and down-regulated the expression of Ia(b). The capacity to stimulate alloreactive T cells and the production of IL-12p70 by DC were significantly augmented by VIP when compared with VIP-untreated DCs. These data suggest that VIP could promote the phenotypic and functional maturation of DCs, hereby regulating the type and outcome of the conducting immune response.     

Lead effects on development and function of bone marrow-derived dendritic cells promote Th2 immune responses

Although lead (Pb) has significant effects on the development and function of macrophages, B cells, and T cells and has been suggested to promote allergic asthma in mice and humans, Pb modulation of bone marrow (BM)-derived dendritic cells (DCs) and the resultant DC effects on Th1 and Th2 development have not been examined. Accordingly, we cultured BM cells with murine granulocyte macrophage-colony stimulating factor (mGM-CSF)+/-PbCl(2). At day 10, culture supernatant (SN) and non-adherent cells were harvested for analysis. Additionally, day 10 non-adherent BM-DCs were harvested and recultured with mGM-CSF+LPS+/-Pb for 2 days. The day 10 Pb exposure significantly inhibited BM-DC generation, based on CD11c expression. Although fewer DCs were generated with Pb, the existing Pb-exposed DCs had significantly greater MHC-II expression than did the non-Pb-exposed DCs. However, these differences diminished upon LPS stimulation. After LPS stimulation, CD80, CD86, CD40, CD54, and MHC-II were all up-regulated on both Pb-DCs and DCs, but Pb-DCs expressed significantly less CD80 than did DCs. The CD86:CD80 ratio suggests a Pb-DC potential for Th2 cell development. After LPS stimulation, IL-6, IL-10, IL-12 (p70), and TNF-alpha levels significantly increased with both Pb-DCs and DCs, but Pb-DCs produced significantly less cytokines than did DCs, except for IL-10, which further supports Pb-DC preferential skewing toward type-2 immunity. In vitro studies confirm that Pb-DCs have the ability to polarize antigen-specific T cells to Th2 cells. Pb-DCs also enhanced allogeneic and autologous T cell proliferation in vitro, and in vivo studies suggested that Pb-DCs inhibited Th1 effects on humoral and cell-mediated immunity. The Pb effect was mainly on DCs, rather than on T cells, and Pb's modification of DC function appears to be the main cause of Pb's promotion of type-2-related immunity, which may relate to Pb's enhanced activation of the Erk/MAP kinase pathway.     

Estradiol enhances capacity of TLR-matured splenic dendritic cells to polarize CD4+ lymphocytes into IL-17/GM-CSF-producing cells in vitro

There are little data on modulatory effects of estrogens on rat dendritic cell (DC) responses to inflammatory stimuli, and consequently their ability to activate and polarize CD4+ T lymphocyte-mediated immune responses. Splenic conventional DCs from young female Albino Oxford rats were activated in vitro with LPS (TLR4 agonist) or R848 (TLR7/8 agonist) in the presence and absence of 17β-estradiol (E2), and their allostimulatory and CD4+ lymphocyte polarizing ability in mixed leukocyte culture (MLC) were studied. Irrespective of the E2 presence, LPS and R848 up-regulated the expression of MHC II on DCs, so they exhibited enhanced allostimulatory capacity in co-culture with CD4+ lymphocytes. On the other hand, E2 promoted stimulatory action of both TLRs on OX62+ DC IL-23 production, augmented their stimulatory effects on IL-6 and IL-1β production, but diminished their enhancing effects on the expression IL-10 and IL-27 by DCs. Consequently, in MLC, OX62+ DCs activated/matured in the co-presence of E2 and either LPS or R848 increased the levels of IL-17, the signature Th17 cell cytokine, when compared with those activated/matured in the absence of E2. GM-CSF levels were also increased in these MLC. Given that the expression of IL-7 mRNA was diminished in DCs activated/matured in the co-presence of E2 and TLR, this increase most likely did not reflect enhanced differentiation of Th cells producing GM-CSF only (Th-GM).ConclusionsE2 augments capacity of LPS- and R848-activated/matured DCs from young rat spleen to induce differentiation of IL-17- and GM-CSF-producing cells.     

Immunization of retrovirus-transfected dendritic cells induces specific cytotoxic T lymphocytes for two distinct malarial peptides presented by Kd molecule

Plasmodium yoelli sporozoite surface protein 2 (pySSP2) is considered as an important antigen for protection studies in malaria vaccine development. For the liver stage protection, anti-pySSP2 cytotoxic T lymphocyte (CTL) activity in BALB/c mice was investigated by immunization of genetically engineered bone marrow-derived dendritic cells (DCs) expressing pySSP2 peptides. Retrovirus-transfected bone marrow cells cultured with GMCSF and IL-4 for 7 days demonstrated 70-80% of DCs with high CD11c, CD80, CD86, and MHC class I (I-Kd) expression. Dividing bone marrow cells were infected with retrovirus expressing SSP2 on fifth, sixth, and seventh days of culture by prolonged centrifugation for 1 h at 32 degrees C. Transfection efficacy of DCs was assessed using retrovirus-shuttled green fluorescence vector (pMSCV-EGFP neo). A total of 64% of CD11c positive transfected DCs showed green fluorescence. The degree of SSP2 expression in transfected DCs was assessed by immunoprecipitation with SSP2 antibody. Both SSP2 and EGFP transfected DCs had prolonged expression of the engineered gene until day 6 since the transfection. Antigen presentation to nai;ve CTLs was assessed by immunization of retrovirus-infected DCs into BALB/c mice. Kd restricted, antigen-specific two new MHC class I (I-Kd) binding motifs were identified (A and C) in pySSP2 protein. Both A and C induced peptide-specific, IFN-gamma-secreting cytolytic CTLs upon antigen recognition on target cells. Taken together, these data indicate that genetically modified DCs by prolonged centrifugation is effective in enhanced antigen presentation. Immunization of DCs encoding SSP2 gene resulted in identification of two K(d) restricted CTL epitopes and induction of IFN-gamma-secreting cytolytic CD8+ T cells.     

分子嵌合MHC-Ⅰ基因对小鼠DC反应的研究

目的:构建分子嵌合主要组织相容性复合体(MHC)-Ⅰ基因小鼠骨髓造血干细胞,并探讨其诱导脾脏T细胞对异基因小鼠树突状细胞(DC)反应的机制。方法:密度梯度法分离培养BALB/c小鼠骨髓造血干细胞。构建携带C57BL/6小鼠MHC-Ⅰ基因慢病毒载体(病毒感染组),携带无意义基因慢病毒载体(阴性对照组)。分别感染BALB/c小鼠骨髓造血干细胞,构建分子嵌合细胞。分别取病毒感染组、阴性对照组及未加入病毒的空白对照组造血干细胞输注BALB/c小鼠后7d,获取脾脏T淋巴细胞,分别与C57BL/6小鼠DC进行混合淋巴细胞培养,测定刺激指数。结果:成功体外分选及培养BALB/c小鼠骨髓造血干细胞。病毒感染组C57BL/6小鼠MHC-Ⅰ蛋白表达率可达98.17%。单向混合淋巴细胞培养结果显示,C57BL/6小鼠DC对输注病毒感染组细胞后BALB/c小鼠脾脏T细胞刺激指数明显降低(P<0.01)。结论:输注分子嵌合MHC-Ⅰ基因造血干细胞后,小鼠脾脏T细胞对异基因小鼠DC反应明显减低。     

Novel immunomodulatory effects of adiponectin on dendritic cell functions

Adiponectin (ADN) is an adipocytokine with anti-inflammatory properties. Although it has been reported that ADN can inhibit the immunostimulatory function of monocytes and macrophages, little is known of its effect on dendritic cells (DC). Recent data suggest that ADN can regulate immune responses. DCs are uniquely specialised antigen presenting cells that play a central role in the initiation of immunity and tolerance. In this study, we have investigated the immuno- modulatory effects of ADN on DC functions. We found that ADN has only moderate effect on the differentiation of murine bone marrow (BM) derived DCs but altered the phenotype of DCs. The expression of major histocompatibilty complex class II (MHCII), CD80 and CD86 on ADN conditioned DCs (ADN-DCs) was lower than that on untreated cells. The production of IL-12p40 was also suppressed in ADN-DCs. Interestingly, ADN treated DCs showed an increase in the expression of the inhibitory molecule, programmed death-1 ligand (PDL-1) compared to untreated cells. In vitro co-culture of ADN-DCs with allogeneic T cells led to a decrease in T cell proliferation and reduction of IL-2 production. Concomitant with that, a higher percentage of CD4(+)CD25(+)Foxp3(+) regulatory T cells (Tregs) was detected in co-cultures of T cells and ADN-DCs. Blocking PD-1/PDL-1 pathway could partially restore T cell function. These findings suggest that the immunomodulatory effect of ADN on immune responses could be at least partially be mediated by its ability to alter DC function. The PD-1/PDL-1 pathway and the enhancement of Treg expansion are implicated in the immunomodulatory mechanisms.     

Phthalate esters modulate the differentiation and maturation of mouse peripheral blood mononuclear cell-derived dendritic cells

Phthalate esters, such as di‐(2‐ethylhexyl) phthalate (DEHP), are widespread environmental contaminants. Previously, we have observed that DEHP exacerbates dermatitis elicited by mite antigen in NC/Nga mice. Also, DEHP enhances the functions of bone marrow‐derived dendritic cells (DCs) in vitro. The present study sought to investigate whether phthalate esters affect peripheral blood mononuclear cell (PBMC)‐derived DCs of NC/Nga mice. First, we studied the time course of DC generation from PBMCs and the dose dependency of granulocyte macrophage colony‐stimulating factor and interleukin‐4, and then determined the conditions under which DC differentiation and maturation are moderately induced from PBMCs. Under the conditions determined above, DEHP at 10 μ m significantly inhibited the expression of DC differentiation and maturation markers, such as CD11c, CD40, CD80, CD86 and CD205, whereas mono‐(2‐ethylhexyl) phthalate, a metabolite of DEHP, did not. Furthermore, the effects of DEHP on PBMC‐derived DCs were partially rescued by treatment with ICI 182,780, an estrogen receptor antagonist. Taken together, these results suggest that DEHP can modulate the differentiation and maturation of mouse PBMC‐derived DCs at least partially through activation of the estrogen receptor under our experimental conditions.