氟比洛芬酯对食管癌患者围术期外周血树突状细胞亚群数量的影响

目的：观察氟比洛芬酯对食管癌患者围术期外周血中天然树突状细胞（ dendritic cells,DCs）亚群数量的影响。方法纳入健康志愿者20例和拟行开胸食管中段癌根治切除术患者40例（对照组20例、干预组20例）。干预组术前（切皮前30 min）给予氟比洛芬酯（2 mg· kg－1）,术毕（缝皮前30 min）给予氟比洛芬酯（2 mg· kg－1）,对照组术前、术毕给予脂肪乳。两组患者术后均使用舒芬太尼＋昂丹司琼行自控静脉镇痛（ patient－controlled intravenous analgesia, PCIA）。流式细胞术检测术前、术毕24、48 h外周血中髓样DC（ myeloid－DCs, mDCs）和浆细胞样DC（ plasmacytoid－DCs, pDCs）两个主要亚群在外周血单个核细胞（ peripheral blood mononuclear cells, PBMCs）中所占比例及其变化。记录术后48 h内的视觉模拟评分（ visual analogue scale, VAS）和舒芬太尼消耗量。结果与健康志愿者相比,食管癌患者外周血mDCs和pDCs亚群在PBMCs中所占比例均显著降低。与对照组相比,干预组患者使用氟比洛芬酯后,术毕24 h外周血中mDCs和pDCs亚群数量显著升高;术毕48 h对照组和干预组患者外周血中mDCs、pDCs亚群数量无显著差异。术毕12、24、36、48 h的对照组和干预组患者VAS评分无显著差异;与对照组相比,干预组患者术毕48 h消耗的舒芬太尼显著减少。结论使用氟比洛芬酯进行围术期镇痛时可以一定时间内上调食管癌患者外周血中DCs亚群水平。     

慢性特发性血小板减少性紫癜患者树突状细胞亚群改变及临床意义

目的探讨树突状细胞(DC)在慢性特发性血小板减少性紫癜(ITP)发病中的作用及糖皮质激素治疗对外周血DC亚群的影响。方法四色荧光流式细胞术分析35例ITP患者糖皮质激素治疗前、治疗后及20名健康人(对照组)外周血DC前体细胞亚群pDC1/pDC2(CD11c+CD123-/CD11c-CD123+)比值。结果35例ITP患者中,6例外周血pDC缺乏,其余29例患者外周血pDC1/pDC2比值为0.77±0.16,低于对照组(1.94±0.44)(P<0.05);治疗后无外周血pDC缺乏患者,血小板计数>100×109/L者27例,pDC1/pDC2比值为1.83±0.43,较治疗前显著升高(P<0.05),与对照组比较差异无统计学意义(P>0.05);血小板计数<100×109/L者8例,pDC1/pDC2比值为1.17±0.24,较治疗前显著升高(P<0.05),但与对照组比较,仍显著降低(P<0.05)。结论慢性ITP患者外周血pDC数量缺乏或pDC1相对减少;糖皮质激素治疗可提升患者外周血pDC数量,完全或部分纠正DC亚群失衡。     

慢性阻塞性肺疾病急性加重期患者外周血中树突状细胞亚群的水平变化及其意义

目的 探讨慢性阻塞性肺疾病急性加重期(AECOPD)患者外周血中树突状细胞(DC)亚群的水平变化及其意义.方法 选取该院呼吸内科收治的120例AECOPD患者、门诊复查的60例COPD稳定期(SCOPD)患者和门诊体检的60例健康人作为研究对象,所选研究对象年龄、性别基本相匹配.采取流式细胞术检测各组研究对象外周血DC亚群、测定肺功能指标及相关炎性因子水平,并分析DC亚群与AECOPD患者肺功能指标的关系.结果 AECOPD患者的第1秒用力呼气容积(FEV1％)、FEV1/FVC(％)值比较发现慢性阻塞性肺疾病全球倡议(GOLD)Ⅰ组>GOLDⅡ组>GOLDⅢ ～Ⅳ组,且差异有统计学意义(P<0.05).AECOPD及SCOPD患者的C-反应蛋白(CRP)、血沉(ESR)、白细胞计数(WBC)值、mDCs、pDCs细胞比例均高于正常对照组,而CD123+、CD11+细胞比例均低于正常对照组,均差异有统计学意义;AECOPD患者的CRP、ESR、WBC值及mDCs、pDCs细胞比例组间比较发现GOLDⅠ组GOLDⅡ组>GOLDⅢ ～Ⅳ组,均差异有统计学意义(P<0.05).AECO-PD患者的FEV1％值与mDCs、pDCs细胞比例值呈显著的负相关(P<0.05);AECOPD患者的FEV1％值与CD123+、CD11+细胞比例值呈显著的正相关(P<0.05).结论 AECOPD患者的DC细胞亚群水平显著的改变,并且与患者的疾病严重程度具有一定的关系,提示DC细胞亚群与患者炎性反应过程有关.     

肿瘤患者外周血树突状细胞亚群的变化及意义

目的 探讨肿瘤患者外周血树突状细胞亚群的变化及临床意义.方法 采用流式细胞术检测23例肿瘤患者和19例健康人外周血CD11c 、CD123 的树突状细胞亚群和淋巴细胞的数量.结果 正常对照组外周血CD11c DC占白细胞比例为0.19%±0.08%,绝对数为(9.3±3.9)× 106个/L;CD12 3 DC点白细胞比例为0.1 5%±0.07%,绝对数为(7.5±3.5)× 106个/L.肿瘤患者组外周血CD123 DC占白细胞比例为0.09%±0.06%,绝对数为(4.7±3.1)×106个/L;CD123 DC占白细胞比例为0.10%±0.06%,绝对数为(5.3±3.2)× 106个/L.与正常对照组比较,肿瘤患者CD11cDC比例和绝对数显著下降(P＜0.05);而CD123 DC比例和绝对数下降无显著性差异.另外,与正常对照组比较,肿瘤患者外周血T、Th、Ts、NK细胞数增均减低(P＜0.05);B细胞绝对数无显著差异.结论 肿瘤患者外周血CD1c DC数量下降,伴随T、Th、T s、NK细胞数降低,可能与肿瘤的发生发展有关.     

树突状细胞及TLR2,TLR9表达在溃疡性结肠炎中的作用研究

目的:观察溃疡性结肠炎(UC)患者两个亚群外周血树突状细胞(DC)即髓样树突状细胞(m DC)和浆样树突状细胞(p DC)的相对数量及外周血m DC表达Toll样受体2(TLR2)、p DC表达Toll样受体9(TLR9)的水平,探讨其在UC中的作用。方法:分离UC患者及健康对照组人群外周血单个核细胞(PBMC),经流式细胞仪分别检测外周血m DC和p DC占PBMC的比例;经流式细胞仪分别检测外周血m DC表达TLR2的平均荧光强度以及p DC表达TLR9的平均荧光强度。结果:UC患者外周血m DC占PBMC的比例低于健康对照组,差异有统计学意义(P<0.01);UC患者外周血p DC占PBMC的比例低于健康对照组,差异有统计学意义(P<0.01);UC患者外周血m DC表达TLR2的平均荧光强度低于健康对照组,差异有统计学意义(P<0.05);UC患者外周血p DC表达TLR9的平均荧光强度低于健康对照组,差异有统计学意义(P<0.05)。结论:UC患者外周血m DC,p DC频率及m DC表达TLR2,p DC表达TLR9的水平均有不同程度下降,提示其在UC发病中可能起重要作用。     

活动性肺结核合并糖尿病患者血糖水平与树突状细胞亚群及T细胞亚群的相关性研究

目的 研究活动性肺结核合并糖尿病患者血糖水平与树突状细胞(DCs)亚群及T细胞亚群的相关性。方法 选取2020年1月至2022年1月惠州市中心人民医院接受诊治的200例患者为研究对象,按照病情分为单纯活动性肺结核患者(对照组)和活动性肺结核合并糖尿病患者(研究组),每组各100例。按照患者糖化血红蛋白(HbA1c)水平将研究组患者分为轻度高血糖组(10 mmol/L≤HbA1c<13.9 mmol/L)45例、中度高血糖组(13.9 mmol/L≤HbA1c<22 mmol/L)33例、重度高血糖组(HbA1c≥22 mmol/L)22例。比较每组患者血糖指标、DCs亚群、T细胞亚群、细胞因子水平,并分析不同血糖程度与DCs、T细胞亚群的相关性。结果 四组患者各血糖指标比较为对照组<轻度高血糖组<中度高血糖组<重度高血糖组(P<0.05);DCs亚群(cDC1、cDC2、pDC)及T细胞亚群(CD4⁺、CD8⁺、CD3⁺)分布值比较为对照组>轻度高血糖组>中度高血糖组>...     

树突状细胞分泌的IFN-λ在丙型肝炎病毒感染中的作用

近年来在Ⅲ型干扰素(IFN-λ)基因区域的单核苷酸多态性可以很好地预测自发性的或是经过药物治疗的丙型肝炎病毒(HCV)感染患者的愈后。至此,越来越多的证据表明,人体先天性免疫应答与IFN-λ基因型效应息息相关。树突状细胞(DCs)在宿主HCV感染的免疫应答中起到关键作用,并且这些至关重要的作用在IFN-λ基因型效应中也正逐渐体现。DCs可以分为很多个亚群,其中主要分泌IFN-λ的DCs,有骨髓样树突状细胞和浆细胞样树突状细胞。基于DCs生物学的复杂性,本综述旨在总结目前已知的有关DCs在HCV感染中所起到的作用以及IFN-λ与DCs的关系。     

特应性皮炎患儿外周血DC1／DC2亚群的检测及临床意义

目的:探讨特应性皮炎(AD)患儿外周血树突状细胞亚群(DC1/DC2)的其临床意义。方法:采用免疫荧光三标记流式细胞术检测38例AD患儿外周血DC亚群(DC1/CD11c+和DC2/CD123+),并以35例正常儿童作为对照。结果:AD患儿外周血CD123+DC百分率(1.28±0.59)%,比对照组(0.69±0.25)%明显增高(P<0.05),CD11c+DC百分率(2.81±1.44)%与对照组(2.56±0.78)%相比,差异无显著性(P>0.05);AD患儿CD11c+DC和CD123+DC与疾病严重程度无关(P>0.05)。结论:特应性皮炎患儿外周血中DC2亚群占优势,导致Th1/Th2细胞向Th2类方向偏移,可能与特应性皮炎的发病机制有关。     

树突状细胞前体细胞亚群四色荧光分析方法探讨

目的建立树突状细胞(DC)前体细胞(pDC)亚群四色荧光检测方法,探讨以Lin-人类白细胞抗原(HLA)-DR+细胞群和以CD34-Lin-HLA-DR+细胞群设门检测不同来源标本pDC亚群的适用性。方法以Lin-HLA-DR+细胞群和以CD34-Lin-HLA-DR+细胞群设门分别检测健康成人外周血、脐带血、骨髓、粒细胞集落刺激因子(G-CSF)动员外周血pDC亚群各15例,比较两种设门方法的pDC检测结果。结果以该两种方法设门,pDC1/pDC2比值:外周血=脐带血>骨髓>G-CSF动员外周血;对同一种标本,两种设门方法之间比较,pDC1/pDC2比值差异无统计学意义;外周血CD34-Lin-HLA-DR+/单个核细胞(MNC)与Lin-HLA-DR+/MNC比值、pDC/CD34-Lin-HLA-DR+比值与pDC/Lin-HLA-DR+比值差异无统计学意义;而脐带血、骨髓、G-CSF动员外周血的CD34-Lin-HLA-DR+/MNC比值均显著性低于Lin-HLA-DR+/MNC比值,pDC/CD34-Lin-HLA-DR+比值显著性高于pDC/Lin-HLA-DR+比值。结论检测pDC亚群,对一般外周血标本,以Lin-HLA-DR+细胞群设门或以CD34-Lin-HLA-DR+细胞群设门,均可取得理想结果;对CD34+细胞含量相对丰富的脐带血、骨髓、G-CSF动员外周血等的标本,以CD34-Lin-HLA-DR+细胞群设门能明显优化检测结果。     

HBsAg多肽刺激的树突状细胞治疗慢性乙型肝炎的临床效果

目的 探讨体外诱导自体HBsAg多肽刺激的树突状细胞(DCs)治疗慢性乙型肝炎(CHB)的作用.方法 治疗组CHB患者32例,应用常规保肝治疗并皮下注射回输体外诱导自体HBsAg多肽刺激的DCs,治疗后3个月时评估疗效.同期CHB患者30例作为对照组.结果 治疗组T淋巴细胞亚群比例、肝功能指标、HBeAg阴转率和HBeAb阳转率均较对照组明显改善,HBVDNA阴转率无明显差异.治疗组HBV DNA下降量和CD4+、CD8+细胞水平明显高于对照组.结论 回输自体HBsAg多肽刺激的DCs可能通过调节免疫耐受,起到抑制乙型肝炎慢性化的效果.     

Modifying toll-like receptor 9 signaling for therapeutic use

Toll-like receptor (TLR) 9 recognizes synthetic oligodeoxynucleotides (ODN) containing unmethylated deoxycytidyl-deoxyguanosine (CpG) motifs and mimics the immunostimulatory activity of bacterial DNA. Both innate and adaptive immune systems are activated through TLR9 signaling and thus its synthetic agonists or inhibitors have potential significance as a target for therapeutic use in immunological disorders. Interestingly, TLR9 found in the dendritic cells and B cells produce differential outcome in response to structurally distinct CpG-ODNs. While one class of CpG-ODN activates B cells and produce immunoglobulin, other can either redirect plasmacytoid dendritic (pDC) cells to secrete high level of IFNalpha or myeloid dendritic cells (mDC) to produce Th1-like cytokines and chemokines necessary for asthma control. This review focuses on potential use of various synthetic CpG to modify TLR9 signaling for therapeutic treatment of multiple diseases including cancer, asthma, allergy and systemic lupus erythematosus (SLE).     

应用自体细胞因子诱导杀伤细胞治疗肝硬化对肝功能影响的研究

目的观察自体细胞因子诱导杀伤细胞（CIK细胞）治疗慢性HBV DNA阳性肝硬化患者对肝功能的影响。方法CIK细胞在体外培养前后以及回输体内后检测CD3^＋、CD3^＋CD4^＋、CD3^＋CD8^＋、CD3^＋、CD56^＋细胞以及mDC和pDC。治疗肝硬化患者,比较治疗前后病毒学指标及肝脏功能的变化。结果培养结束以及回输体内后,CD3^＋细胞、CD3^＋CD8^＋细胞、CD3^＋、CD56^＋细胞细胞较培养前显著升高,mDC和pDC在回输后也明显增高,肝脏功能较治疗前有所好转。所有患者均能耐受治疗。结论CIK细胞可明显提高免疫效应细胞数量,具有一定的抗病毒效应作用,毒副作用低。没有加重肝脏损伤,而对于肝功能有一定的改善作用。     

自体细胞因子诱导杀伤细胞治疗肝硬变的临床疗效研究

目的观察自体细胞因子诱导杀伤细胞（CIK细胞）治疗HBVDNA阳性肝硬化患者的近期疗效。方法CIK细胞在体外培养前后以及回输体内后检测CD3^＋、CD3^＋CD4^＋、CD3^＋CD4^＋、CD3^＋、CD56^＋、CD25^＋细胞以及mDC和pDC。治疗肝硬化患者,比较治疗前后病毒学指标及肝脏功能的变化。结果培养结束以及回输体内后,CD3^＋细胞、CD3^＋CD8^＋细胞、CD3^＋CD8^＋细胞较培养前显著升高,mDC和pDC在回输后也明显增高,肝脏功能较治疗前有所好转。所有患者均能耐受治疗。结论CIK细胞可明显提高免疫效应细胞数量,具有一定的抗病毒效应作用,毒副作用低。     

树突状细胞疫苗注射治疗慢性乙型肝炎后外周血T细胞亚群的变化

目的研究轻度慢乙肝患者经树突状细胞疫苗治疗后外周血中T细胞亚群的改变。方法32例轻度慢乙肝患者,每次注射>106乙肝疫苗冲击树突状细胞,1次/月,共12次。于治疗前后利用流式细胞仪测定患者外周血中T细胞亚群。结果在树突状细胞疫苗治疗后有应答反应的20例患者中可见CD4+T细胞和CD8+CD28+T细胞明显增加,而CD8+T细胞和CD8+CD28-T细胞显著降低。结论树突状细胞疫苗有可能活化慢乙肝患者的免疫反应,对其治疗有一定疗效。     

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。     

Effects of Pravastatin on the Function of Dendritic Cells in Patients with Coronary Heart Disease

Abstract: The aim of the study was to investigate the functional profile of dendritic cells in patients with coronary heart disease and the effects of pravastatin on this. Forty‐eight patients with coronary heart disease were divided into three groups: 16 treated with pravastatin 10 mg/day, 16 treated with pravastatin 20 mg/day and 16 not treated with pravastatin. Dendritic cells from 48 patients with coronary heart disease (before and 4 weeks after the treatment) and 16 healthy individuals were derived from peripheral blood. CD86 of dendritic cells was assessed by flow cytometry. Immunostimulatory capacity of dendritic cells was measured by mixed lymphocyte reaction. The levels of cytokines in the medium of mixed lymphocyte reaction were analysed. Blood lipids and high‐sensitivity C‐reactive protein were measured. Compared to normal group, more CD86+ dendritic cells were expressed in coronary heart disease and greater immunostimulatory capacity of dendritic cells in coronary heart disease was demonstrated. T lymphocytes in coronary heart disease in mixed lymphocyte reaction secreted higher levels of pro‐inflammation cytokines and lower levels of anti‐inflammation cytokines. CD86 expression significantly correlated with C‐reactive protein, but did not correlate with low‐density lipoprotein cholesterol. Both dosages of pravastatin markedly inhibited the function of dendritic cells and lowered C‐reactive protein, which is independent of plasma cholesterol lowering. The anti‐inflammatory effect of pravastatin showed no obvious difference between the two dosage groups. In conclusion, dendritic cells were activated in coronary heart disease and dendritic cell‐mediated immune mechanisms may be involved in the pathogenesis of coronary heart disease. Pravastatin can inhibit dendritic cell activation, which is independent of plasma cholesterol lowering. Pravastatin in different doses showed no apparent differences in the inhibition of dendritic cell functions.     

Affects of immunosuppression on circulating dendritic cells: an adjunct to therapeutic drug monitoring after heart transplantation

OBJECTIVES: Recent evidence emerges dendritic cells (DCs) as pharmacological targets of immunosuppressive drugs. Therefore, in this study we monitored DCs in peripheral blood to compare the effects of calcineurin inhibitors (CNI: cyclosporine, tacrolimus) and mammalian target of rapamycin inhibitors (sirolimus, SRL, everolimus, ERL) basis-immunosuppressive therapies in human heart transplanted (HTx) recipients. METHODS: We compared HTx recipients which were converted from either CNI to ERL (severe renal dysfunction, n=8), or from SRL to ERL (approval of ERL for HTx, n=8) with 20 healthy human controls. Twenty four after the last CNI or SRL dose recipients were treated with ERL/BID on days 1-3. Peripheral blood was collected at trough in the morning before and on day 4 after conversion. Percentages of positive myeloid and plasmacytoid DC (m and pDC) subsets in peripheral blood were analysed by flow cytometry. The status of maturation was further characterised by flow cytometry analysis of % expression of CD83 and % expression of various intracellular cytokines (IL-1beta, TNF-alpha, IL-8, IL-12), respectively. RESULTS: HTx recipients had higher % positive mDCs regardless the immunosuppressive therapy compared to controls (p<0.05). Whereas, % positive pDCs were only significantly lower in recipients converted from CNI to ERL compared to controls (p<0.05). The data consolidate the finding that the subset ratio pDCs/mDCs was lower in recipients compared to controls. But after conversion from CNI or SRL to ERL the ratio increased towards pDCs. Percentages of expression of CD83 on mDCs were not different among the recipient groups and controls. Recipients with CNI and SRL had higher % expression of IL-12 and lower % expression of IL-1beta compared to controls (p<0.05). However, after conversion to ERL % expression of both IL-12 and IL-1beta returned to control values in both groups. CONCLUSIONS: The results showed that analysis of immunosuppression of circulating DCs in peripheral blood may be an adjunct to therapeutic drug monitoring to optimize immunosuppressive therapy after HTx.