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1.
目的探讨Bryostatin-1对树突状细胞(DC)免疫功能的调节作用。方法联合应用GM-CSF和IL-4自人外周血单核细胞定向分化DC;以Bryostatin-1刺激TNF-α诱导成熟的DC,收集DC及其上清夜,以FACS和ELISA方法分析DC表面免疫分子(CD80、CD83、CD86、HLA-DR和B7-H1)和细胞因子(IFN-γ、IL-1β、IL-10和IL-12)表达水平;自DC提取RNA,以Northern blot分析DC的B7-h1 mRNA表达水平;以DC为诱导细胞,进一步检测DC的免疫诱导功能。结果Bryostatin-1通过降低DC表面B7-H1表达和增强B7.2表达,以及调节DC细胞因子(IL-1b、IL-12和IFN-γ)分泌,增强DC的免疫诱导功能,PKC通道特异性的抑制剂BI明显逆转Bryostatin-1的上述作用。结论Bryostatin-1增强DC免疫功能,其机制可能是激活PKC通道。  相似文献   

2.
用Ficoll密度离心及贴壁法获得外周血单个核细胞 (PBMC ) ,PBMC经细胞因子组合诱导分化成树突状细胞 (DC ) :GM CSF (10 0ng/ml)与IL 4 (5 0ng/ml)诱导 5d后 ,分别加入TNF α (10ng/ml)或rhsCD4 0L (2 μg/ml)继续培养 4d ;倒置显微镜下观察DC形态 ,免疫荧光标记和流式细胞术分析DC表型 (CD1a、CD80、CD83、HLA DR、CD14、CD16、CD19)及摄取FITC Dextran抗原的能力 ;3 H TdR掺入法检测DC刺激自体混合淋巴细胞体外增殖反应 (MLR )能力 ;ELISA法分析DC培养上清中IL 12的水平 ;Trans well细胞趋化实验检测DC对自体外周T淋巴细胞的趋化能力。发现经rhsCD4 0L刺激的DC表面分子 (CD1a、CD80、CD83、HLA DR )的表达水平高于经典的细胞因子组合组 (GM CSF +IL 4 +TNF α ) ,同时rhsCD4 0L刺激后的DC摄取FITC Dextran的能力下降而刺激自体MLR和分泌IL 12的能力明显提高 ;而且rhsCD4 0L诱导的DC表面趋化因子受体CXCR4的表达水平及对自体外周T淋巴细胞的趋化能力均强于TNF α或FL激发的DC。rhsCD4 0L在体外不仅具有显著的诱导DC分化 ,促进DC成熟的功能 ,而且经rhsCD4 0L作用的DC能更有效地激发T淋巴细胞  相似文献   

3.
目的 体外扩增慢性乙肝病人的树突状细胞 (dendriticcell,DC) ,从细胞表型和功能上鉴定和研究。方法 用含GM CSF和IL 4的无血清培养基AIM V体外培养慢性乙肝病人外周血单个核细胞 ,获得树突状细胞。流式细胞仪检测细胞表型 ,IL 1 2ELISA试剂盒检测DC分泌IL 1 2的水平 ,并观察加入细胞因子TNF α后对DC培养的影响。结果 慢性乙肝病人的外周血单个核细胞用AIM V培养及细胞因子诱导后 ,经贴壁法纯化 1 0 0mL可获得 0 .5× 1 0 7~ 1 .5× 1 0 7成熟的具有典型形态的DC ,加入TNF α后 ,CD83阳性占 60 .80 % ,明显高于未加组 (P <0 .0 5) ,IL 1 2分泌较未加TNF α组增高近 1 0倍。结论 ①慢性乙肝病人的DC可用AIM V无血清培养基及特定的细胞因子诱导在体外大量获得 ,TNF α是诱导DC成熟的重要的细胞因子。②典型的细胞形态和CD1 4 -、HLA DRhigh+、CD86high+的细胞表面分子特征可作为临床上快速鉴定培养DC的标志。  相似文献   

4.
目的 研究IL-24基因修饰的树突状细胞(DC)与同源细胞因子诱导的杀伤细胞(cyto-kine-induced killer,CIK)共培养后对A549肺癌细胞的杀伤作用及其机制.方法 从健康人外周血单个核细胞中常规诱导DC、CIK细胞,同时抽提重组腺病毒质粒pAdEasy-1-pTrack-CMV-IL-24,Pac I酶切线性化后脂质体转染QBl-293A细胞,收获Ad-IL-24重组病毒.将IL-24基因通过重组病毒导入已负载肿瘤抗原的DC,获得细胞称为DC-IL-24.RT-PCR和ELISA法检测DC中IL-24基因的表达,FCM和ELISA法检测DC表型及分泌细胞因子能力的变化,将DC和CIK细胞混合培养,溶血试验检测CIK细胞产生穿孔素的能力,FCM法检测共培养的DC-CIK细胞对A549肺癌细胞细胞毒活性的变化.结果 获得了高滴度的重组病毒Ad-IL-24并成功将IL-24基因导入DC,在倒置荧光显微镜下可观察到荧光,IL-24可上调Dc表而CD80、CD83、HLA-DR、CD40、CXCR4分子的表达,转染后Dc分泌IL-12、TNF-α和IL-24的能力显著增强,DC-IL-24更能促进CIK细胞产生穿孔素,与同源CIK细胞共培养后对A549肺癌细胞的细胞毒活件明显增强.结论 IL-24基因修饰的DC能增强自体CIK细胞产生特异性抗肿瘤免疫,其机制与IL-24能促进DC表型成熟,分泌Th1型细胞因子,维持DC活化状态,进而促进CIK细胞活化密切相关.  相似文献   

5.
目的探讨负载自体肿瘤抗原的树突状细胞(DC)联合细胞因子诱导的杀伤细胞(CIK细胞)在晚期肾癌治疗中的临床疗效,并监测患者的免疫状态及不良反应。方法回顾分析2011-01/2013-12期间在我科治疗的82例晚期肾癌患者,单采其外周血单个核细胞(PBMC),其中贴壁细胞经体外诱导产生DC,并负载自体肿瘤细胞裂解物(Ag)制备Ag-DC;T淋巴细胞经体外诱导产生CIK细胞,将Ag-DC与CIK细胞共培养制备Ag-DC-CIK瘤苗,检测DC表面分子的表达及IL-12的分泌,监测CIK细胞增殖情况,并将Ag-DC-CIK分次回输给其中41名患者,以41例单纯的CIK细胞治疗作为对照,治疗2疗程后,检测患者外周血细胞因子水平及T细胞亚群变化,结合影像学等检查综合评价疗效,同时观察不良反应。结果负载肿瘤细胞冻融抗原的DC,高表达CD11c、CD83、CD86、HLA-DR表面分子,IL-12的分泌显著增加。Ag-DC与CIK细胞共培养后,可明显刺激CIK细胞的增殖,同时CD3+CD8+细胞及CD3+CD56+细胞的含量明显增加,此外,Ag-DC-CIK瘤苗的临床疗效明显高于单纯CIK细胞治疗组,2组均未发生严重的不良反应。结论晚期肾癌PBMC来源的DC负载自身肿瘤细胞冻融抗原能提高晚期肾癌患者的免疫状态。  相似文献   

6.
探讨慢性乙肝患者树突状细胞(dendritic cells,DC)对CD4+Th细胞亚群分化的影响。分离慢性乙肝患者外周血单个核细胞(PBMC),以rhIL-4(50 ng/ml)、rhGM-CSF(10 ng/ml)和rhTNF-α(100 u/ml)诱导培养DC。以流式细胞仪检测DC表面CD1a、CD83、CD80、CD86、HLA-DR分子表达情况。MTT法检测DC刺激同种异体淋巴细胞增殖能力。免疫磁珠分离外周血CD4+T细胞亚群,PMA+Ionomycin刺激后胞内荧光染色,流式细胞仪检测辅助性T细胞(helper T cell,Th)内特征性细胞因子IFN-γ/IL-4以判断Th1/Th2分化。ELISA法检测DC或Th细胞培养上清中IL-6、IL-12、IFN-γ和IL-4的含量。结果:慢性乙肝患者的DC表达CD1a、CD83、CD80、CD86、HLA-DR分子水平明显低于正常人(P<0.01);培养至第7天,慢性乙肝患者DC分泌的IL-12水平低于正常人(P<0.01),而分泌的IL-6水平增高(P<0.05)。与正常人相比,慢性乙肝患者外周血中Th1细胞占CD4+T细胞的百分比较低(P<0.01),其Th细胞培养上清中IFN-γ的量也较低(P<0.01)。患者DC与同种异体的健康人Th细胞共培养,刺激Th1型细胞因子IFN-γ产生的能力低于正常人(P<0.01)。慢性乙肝患者体内DC功能的异常可能导致了外周血Th1细胞分化不足。  相似文献   

7.
目的探讨并比较不同细胞因子组合、诱导时间对树突状细胞(DC)体外诱导、成熟,分泌细胞因子水平和刺激淋巴细胞增殖能力的影响,为建立DC体外高效、规模化培育体系提供依据。方法分离健康人外周血单个核细胞(PBMC),贴壁法获得DC前体细胞,用白细胞介素4(IL-4)和粒细胞巨噬细胞集落刺激因子(GM-CSF)诱导2 d,将其分为6组。A组加入肿瘤坏死因子α(TNF-α)、IL-1α和前列腺素E2(PGE2);B组加入α干扰素(IFN-α);C组加入脂多糖(LPS);3组细胞继续培育2 d。D~F组细胞先进行1/3体积换液,24 h后,分别再加入上述A~C组的成熟因子,再继续诱导2 d。分别收获各组上清和细胞,进行细胞计数,采用流式细胞术进行表型分析;采用ELISA检测各组DC分泌IL-12p70的水平、采用细胞增殖检测试剂盒刺激淋巴细胞增殖的能力。结果 6组体外诱导体系均能培育出形态学类似DC的细胞,DC数量与纯度(均达到96%以上),各组间无显著性差异。A组与D组比较:CD40、CD80、CD83、CD86共刺激分子表达水平相近,但D组DC分泌IL-12p70水平高于A组;B组与E组比较:E组DC的CD86表达水平显著高于B组,CD80表达量略高于B组,2组DC分泌IL-12p70的水平相近;C组与F组比较:CD40、CD80、CD83、CD86共刺激分子表达水平和分泌IL-12p70的水平均相近。6组DC刺激淋巴细胞增殖水平均相近。三种不同成熟因子(组合)比较:B组DC共刺激分子CD80水平明显低于C组和A组,且IL-12p70分泌水平最低,而C组和A组共刺激分子表达水平与IL-12p70分泌水平相近。结论 TNF-α、IL-1α和PGE-2三种细胞因子联合以及LPS可有效地诱导DC表达高水平免疫共刺激分子、分泌高水平IL-12p70。  相似文献   

8.
目的 探讨腺病毒(Ad)介导E型沙眼衣原体(Ct)主要外膜蛋白(MOMP)基因转染对树突状细胞(DC)的表型及体内免疫功能的影响.方法 用小鼠重组粒细胞一巨噬细胞集落刺激因子(GM-CSF)及白细胞介素4(IL-4)从小鼠骨髓干细胞中诱导培养DC,并用大肠杆菌脂多糖(LPS)促进DC的成熟,流式细胞仪检测DC表型.用不同感染滴度(MOI)的含增强绿色荧光蛋白(EGFP)基因的重组Ad转染DC,荧光显微镜下观察EGFP的表达细胞百分率,选择最佳MOI;用最佳MOI的含MOMP基因的重组腺病毒(Ad-MOMP)转染DC,流式细胞术检测转染前后DC表型变化,并用活细胞计数试剂盒8(CCK-8)检测转染前后DC刺激同种淋巴细胞增殖的能力;ELISA检测转染前后DC分泌细胞因子及DC、T细胞共同培养上清中细胞因子的水平.Ad-MOMP转染DC经尾静脉免疫小鼠,ELISA检测其脾脏淋巴细胞分泌的细胞因子.结果 诱导的DC形态典型,表面高表达CD11c、MHCⅡ类分子,中度表达CD80分子.MOI=1000为重组Ad转染DC最佳滴度,此时90%以上的DC表达荧光,Ad-MOMP转染DC后能检测到MOMP的表达.Ad-MOMP转染对DC表面的特征性表型CD11c无影响,而CD80及MHCⅡ表达上调;转染后的DC分泌大量IL-12,具有较强的刺激同种异体淋巴细胞增殖的能力,并刺激T细胞分泌大量IFN-γ.Ad-MOMP转染DC后尾静脉免疫小鼠,其脾细胞产生高水平的IFN-γ.结论 Ad载体能介导外源MOMP基因在DC中的表达,Ad-MOMP转染DC后MOMP基因的表达能增强DC抗原提呈功能,促进DC活化,转染后的DC与T细胞共孵育能诱导T细胞向TH1细胞分化,体内实验表明Ad-MOMP转染DC能诱导衣原体特异性TH1反应.这为Ad-MOMP转染的DC疫苗用于免疫治疗提供了理论依据和技术基础.  相似文献   

9.
目的 :研究HBsAg冲击的慢性乙肝患者单核细胞来源的树突状细胞 (DCs)的功能状况及体外对HBV特异性CTL的诱导作用 ,初步探讨诱导特异性抗HBV细胞免疫的途径。方法 :分离慢性乙肝患者外周血单核细胞 ,以GM CSF +IL 4 +TNF α培养诱导DCs,加入HBsAg冲击以诱导HBV特异性DCs。采用FCM测定细胞表面免疫分子CD1a、CD83、CD86、CD80、CD4 0以及HLA DR的表达水平 ,ELISA法检测培养上清中细胞因子IL 6、IL 12的分泌含量 ,MTT法测定DC刺激同种异体淋巴细胞增殖的能力 ,LDH法检测DC诱导的患者外周血T细胞对HepG2 2 2 15 (转染HBVDNA)、HepG2肝癌细胞株及K5 6 2白血病细胞株的细胞毒作用。结果 :HBsAg冲击的DC其表达CD1a、CD83、CD86、CD80、CD4 0、HLA DR表面分子明显高于对照组 (P <0 0 1,P <0 0 5 ) ,分泌IL 12的水平也高于对照组 (P <0 0 1) ,而分泌IL 6的水平则较对照组显著降低(P <0 0 1) ;HBsAg冲击的DC刺激同种异体淋巴细胞增殖的能力明显增强 (P <0 0 5 ) ,并可有效地诱导自体CTL对转HBV基因的HepG2 2 2 15细胞高效特异性杀伤作用 (P <0 0 1)。结论 :慢性乙型肝炎患者单核细胞来源的DCs经HBsAg抗原冲击后 ,生物学活性增强 ,并且能有效地诱导对HBV特异性反应的CTL。  相似文献   

10.
目的研究SOCS1沉默的树突状细胞特异性抗肿瘤作用机制,并探讨RNAi技术在喉癌基因治疗中的应用前景,为树突状细胞的临床应用提供新思路和理论依据。方法以细胞因子GM-CSF、IL-4和TNF-α体外诱导扩增外周血单核细胞来源的DC,倒置显微镜下观察DC形态特征;构建RNAi载体转染DC,Western blot检测SOCS1的表达情况,筛选抑制SOCS1表达的有效靶序列;流式细胞术检测DC表面分子CD83、CD86和HLA-DR的表达;ELISA法分析上清中IFN-γ的含量;MTT法评估DC刺激T细胞增殖的能力及诱导细胞毒性T细胞的杀伤活性。结果 DC体外诱导培养成功;设计的RNAi载体经测序验证无误。干扰序列5可显著下调SOCS1表达水平;SOCS1沉默联合喉癌Hep-2抗原致敏的DC可显著上调表面分子标志CD83(85.61±0.96)%、CD86(96.86±1.20)%和HLA-DR(98.02±0.94)%的表达;该组DC能有效刺激T细胞增殖,增加IFN-γ的分泌量,最终增强CTL的特异性杀伤作用,效靶比为50:1时其杀伤活性显著高于对照组(P<0.01)。结论 SOCS1沉默并负载喉癌Hep-2抗原的DC可以产生高效而特异性的抗喉癌免疫应答。  相似文献   

11.
Over 200 schizophrenic patients belonging to three major and interrelated pedigree complexes have been investigated over the past 30 years in a North Swedish geographically isolated population, presently numbering about 6,000. An intensive investigation of a number of biochemical correlates and genetic markers in a few selected families belonging to one of the major pedigrees has indicated new strategies for the current research program.
Schizophrenia, as defined operationally, is significantly associated with decreased activities of two enzymes (1) blood platelet monoamine oxidase, (2) plasma dopamine-β-hydroxylase, and (3) with the genetic marker Gc2 (group specific antigen). Both enzymes are subject to genetic variation. A positive score for linkage between schizophrenia and low plasma DBH activity has been calculated, but, so far, available data are insufficient for discrimination between linkage and partial contribution of genetically controlled low plasma DBH to the pathogenesis of the disease. Alternatively, both mechanisms could be involved.
As a model for continued research, schizophrenia is explained as based on a double dominant-recessive genotype (Aabb), representing a vulnerability which in about 50 % of cases develops into clinical schizophrenia. It is suggested that the dominant mutation (A) operates on or affects MAO activity, and that the recessive genotype (bb) is instrumental in low variates of DBH activity and very likely such variates within the normal range of physiological variation. Moreover, it is suggested that the combined effects of MAO- and DBH-reduced efficiency on the metabolism of e.g. dopamine could be an essential pathogenic mechanism for the schizophrenic illness which is segregating in this population.  相似文献   

12.
HLA-A,-B,-C,-DRB1 and -DQB1 alleles have been studied in Chimila Amerindians from Sabana de San Angel (North Colombian Coast) by using high resolution molecular typing. A frequent extended haplotype was found:HLA-A*24:02-B*51:10-C*15:02-BRB1*04:07-DQB1*03:02 (28.7%) which has also been described in Amerinndian Mayos Mexican population (Mexico, California Gulf, Pacific Ocean). Other haplotypes had already been found in Amerindians from Mexico (Pacific and Atlantic Coast), Peru (highlands and Amazon Basin), Bolivia and North USA. A geographic pattern according to HLA allele or haplotype frequencies is lacking in Amerindians, as already known. Also, five new extended haplotypes were found in Chimila Amerindians. Their HLA-A*24:02 high frequencies characteristic is shared with aboriginal populations of Taiwan; also, HLA-C*01:02 high frequencies are found in New Zealand Maoris, New Caledonians and Kimberly Aborigines from Australia. Finally, this study may show a model of evolutionary factors acting and rising one HLA allele frequency (-A*24:02), but not in others that belong to the same or different HLA loci.  相似文献   

13.
Starting with the integument, we see many organs are contractile sacs or multiples thereof, which tubes or bags constitute the major part of the entire body. Recognition of this basic unit and its characteristics sheds new light, individually and collectively, on many disorders previously considered unrelated. Muscular tears and perforations develop in the walls of these chambers, being no way peculiar to those organs, wherein, hydrochloric acid occurs. So, it is not necessary to explain the absence of excessive acid from patients who exhibit holes in the gastric, uterine, aortic, duodenal, rectal, pulmonary, retina, and other walls. Muscle, not acid is the great common factor relating idiopathic disorders in the gastrointestinal tract to each other and to similar diseases in other systems. When the units are linked together, the lesions tend to appear as arthropathies, i.e. at the joints. Rephrasing common-place observations, frees us from conventional, conceptual cul-de-sacs. An observation is only as good as its interpretation, so all possibilities must be considered, otherwise, we will remain blinded by our misconceptions.  相似文献   

14.
Zusammenfassung Der Einfluß von verschiedenen Nahrungsmitteln auf Methoden zur Bestimmung von Adrenalin (AD), Noradrenalin (NA), Vanillinmandelsäure (VMS), Metanephrinen (MN), Homovanillinsäure (HVS) und 5-Hydroxyindolessigsäure (5-HIE) im 24 h-Harn zur Diagnose des Phäochromozytoms bzw. Karzinoid-Syndroms wurde untersucht. Die in die Untersuchung einbezogenen Nahrungsmittel waren: Tee, Kaffee, Mandeln, Ananas, Käse, Walnüsse, Vanillepudding, Bananen, Tomaten und Milchschokolade. Außerdem wurde der Einfluß des Zigarettenrauchens auf die Bestimmung von AD, NA, VMS und MN untersucht.Walnüsse führten zu einer starken Erhöhung der 5-HIE-Ausscheidung. Bananen erhöhten die Ausscheidung von AD, NA, VMS, MN und 5-HIE. Kaffee und Ananas bewirkten eine geringe Zunahme der MN-Werte. Rauchen von 20–30 Zigaretten/Tag beeinflußte keine der vier Variablen.Wenn die beschriebenen Methoden benutzt werden, sollte lediglich auf den Verzehr von Bananen und Walnüssen vor und während der Harnsammelperioden verzichtet werden, da die oberen Normgrenzen im Harn überschritten werden könnten. Ein Verzicht auf Kaffee und Ananas in normalen Mengen ist nicht erforderlich. Es besteht kein Anlaß, weiterhin die bisherigen umfangreichen Restriktionen der übrigen Nahrungsmittel beizubehalten.  相似文献   

15.
Dimebon, an antihistamine agent, exerts a moderate antianginal effect, improving the function of ischemic focus in the myocardium and decreasing the necrotic zone in experimental myocardial infarction. Dimebon is less active than obsidan, finoptin (except for the size of the necrotic zone), and cordaron. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 122, No. 12, pp. 642–644, December, 1996  相似文献   

16.
17.
Effects of estradiol and testosterone on the intensity of lipid peroxidation and contents of glutathione redox system components in the dermis and epidermis of rat skin were studied. Only estradiol induced considerable dose-dependent and tissue-specific biphasic antioxidant effects on the skin. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 128, No. 12, pp. 663–666, December, 1999  相似文献   

18.
A series of surface-functionalized poly(ether ether ketone) (PEEK) films has been prepared by selective wet-chemistry; they are hydroxylated polymer (PEEK-OH) obtained by reduction, aminated polymer (PEEK-[]-NH2) prepared by coupling a diisocyanate reagent to PEEKOH (PEEK-[]-NCO) followed by hydrolysis, and carboxylated and aminocarboxylated polymers (PEEK-[]-GABA and PEEK-Lysine) resulting from the coupling of aminoacids to PEEK-[]-NCO. The aminated and carboxylated substrata promoted the adhesion and growth of CaCo2 cells in the presence of serum. Fibronectin (FN), an extra-cellular matrix protein, has been covalently fixed and/or adsorbed on various PEEK substrata, in the presence or not of a polymeric surfactant (Pluronic F68). The performances of the FN-grafted substrata (PEEK-[]-FN(1) and PEEK-[]-FN(2)) were significantly higher than those of reference substrata simply coated with FN (PEEK-OH(+FN)(1) and (2), PEEK-[]-NH2(+FN)(1) and (2)), considering the adhesion and spreading of CaCo2 cells in the absence of serum. Moreover, the stability of the adherent cells on the FN-adsorbed substrata dramatically depended on the experimental conditions applied during the PEEK coating with FN.  相似文献   

19.
20.
Homer's epic tale of the 20-year return of Odysseus from the Trojan War is investigated with particular reference to Jung's theory of individuation. Odysseus' meetings with 'the anima' in the form of goddesses, sirens and female monsters and his visit to Hades demonstrate the confrontation and humanization of aspects of the archetypal level of the psyche, central to Jung's theories of psychic growth and development. Jung's important ideas of the psychoid level and the transcendent function are explored and linked both to his investigations into medieval alchemy and with findings from contemporary neuroscience. The importance of Jung's constructive method of the interpretation of dreams and myths is shown to be central.  相似文献   

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