活动性肺结核患者外周血单个核细胞表达TH1/TH2类细胞因子的特点

结核病患者免疫细胞产生TH1类细胞因子的能力低下，而TH2类细胞因子表达各家报道不一。CD4^ T和CD8^ T细胞是结核病免疫中主要的效应细胞，也是细胞因子的主要来源。我们采用流式细胞仪检测CD4^ T和CD8^ T分泌IFN-γ和IL-4的能力，从单细胞水平探讨结核菌感染患者TH1／TH2型免疫反应的特点。     

结核菌素纯蛋白衍生物对哮喘儿童外周血单个核细胞IL-12、IFN-γ/IL-4表达的影响

哮喘涉及的免疫机制复杂,主要表现为Th1/Th2细胞功能失衡,即Th1细胞功能下降和Th2细胞功能亢进。儿童结核菌素反应阳性提示哮喘和变应性发生率降低;随后的研究认为卡介苗及其产物有Th1类免疫调节作用。本文通过观察结核菌素纯蛋白衍化物（PPD）对哮喘患儿外周血单个核细胞（PBMC）产生Th1类细胞因子IL-12和IFN-γ、Th2类细胞因子IL-4的影响,并与植物血凝素（PHA-m）刺激的结果比较,探讨PPD在哮喘发病中的作用及价值。     

口服卡介苗对哮喘小鼠TH1/TH2平衡的影响

卡介苗不仅被应用于结核病的预防,还应用于哮喘、慢支、肿瘤等疾病的治疗,国内外学者以皮内注射卡介苗用于实验性哮喘的治疗取得了很好的疗效。但卡介苗口服用于哮喘防治国内还未见报道。本实验通过口服卡介苗后哮喘小鼠支气管肺泡灌洗液(BALF)和外周血中IL-4、IFN-γ水平的变化,并用流式细胞仪来检测Y_H1/T_H2细胞平衡的影响,进一步探讨哮喘发病机理,为口服卡介苗治疗哮喘提供理论依据。32只BALB/c小鼠分为正常对照组、哮喘组、皮内注射卡介苗 哮喘组、口服卡介苗 哮喘组,以卵蛋白(OVA)致敏激发建     

卡介菌多糖核酸和地塞米松对口腔扁平苔藓TH1/TH2细胞因子的调节作用

口腔扁平苔藓 (orallichenplanus ,OLP)是T淋巴细胞介导的免疫反应 ,其主要病理变化为固有层大量的T淋巴细胞呈带状浸润〔1〕 。卡介菌多糖核酸(BacillusCalmette Guerinpolysaccharidenucleicacid ,BCGPSN)是从卡介菌中经热酚法提取的一种具有免疫调节功能的免疫调节剂 ,对慢性支气管炎、哮喘等疾病有较好的疗效。本研究拟通过观察BCGPSN和地塞米松对OLP患者外周血单个核细胞 (peripheralbloodmononuclearcells ,PBMC)IFN γ和IL 4的调节作用 ,探讨OLP患者的TH1 TH2免疫应答模式及BCGPSN和糖皮质激素对OLP患者TH1 TH2…     

Differences in cytokine production by peripheral blood mononuclear cells (PBMC) between patients with atopic dermatitis and bronchial asthma.

It is widely accepted that type 2 helper T (Th2) lymphocytes play a crucial role in the pathogenesis of atopic dermatitis (AD) as well as bronchial asthma (BA). We measured the amounts of IL-5 and interferon-gamma (IFN-gamma) produced by PBMC upon stimulation with house dust mite (HDM) or Candida albicans (CA) in 17 children (3-15 years) with AD, and compared these values with those of 16 children with BA. Although IL-5 production by PBMC upon stimulation with HDM in patients with AD was significantly higher than that in 13 non-atopic controls (geometric mean = 23.4 pg/ml versus 5.9 pg/ml, P < 0.05), it was significantly lower than that in patients with BA (177.8 pg/ml, P < 0.001). The amount of IL-5 produced by PBMC upon stimulation with CA was also significantly lower in patients with AD than in those with BA (7.2 pg/ml versus 100.0 pg/ml, P < 0.001). The production of IFN-gamma by PBMC stimulated with HDM or CA was also significantly lower in patients with AD than in those with BA (HDM 4. 3 pg/ml versus 12.6 pg/ml, P < 0.05; CA 6.5 pg/ml versus 60.3 pg/ml, P < 0.001). Consequently the ratio of IL-5 to IFN-gamma production was high not only in patients with BA but also in those with AD. These findings suggest that there are some differences in the regulation of in vivo cytokine production between patients with AD and those with BA, although a Th2-dominant profile is common to both.     

HCV感染外周血单个核细胞及其对T淋巴细胞亚群的影响

目的：研究丙型肝炎病毒（ＨＣＶ）感染外周血单个核细胞（ＰＢＭＣ）的情况及其对Ｔ淋巴细胞亚群的影响。方法：运用非同位素原位杂交（ＮＩＳＨ）法和链酶亲和素－生物素（ＳＡＢＣ）法分别检测２０例慢性丙型肝炎患者ＰＢＭＣ中的ＨＣＶ－ＲＮＡ和非结构（Ｎｏｎｓｔｒｕｃｔｕｒａｌ,ＮＳ）蛋白ＮＳ５抗原,同时用ＳＡＢＣ法检测其Ｔ淋巴细胞亚群。结果：８例（４０．０％）患者的ＰＢＭＣ中ＨＣＶ－ＲＮＡ呈构（Ｎｏｎｓｔｒｕ     

羊水来源间充质细胞对于外周单核淋巴细胞增殖抑制作用的机制研究

目的鉴定羊水来源间充质细胞(AF-MCs)并探讨其对于外周单核淋巴细胞(PBMC)增殖活性的影响及机制。方法通过选择性消化和单克隆培养分离及培养人源羊水细胞(AFCs),而后通过核型分析细胞来源和流式分析细胞表型鉴定其为间充质细胞(MCs)。另一方面,将鉴定得到的AF-MCs与PBMC共培养,检测其是否具有免疫抑制功能。运用流式分析CD4~+CD25~+Foxp3~+T细胞频率在共培养前后变化,探讨AF-MCs具有免疫抑制功能的机制。结果AF-MCs来自于胚胎发育过程中的脱落细胞,其具有与骨髓间充质干细胞(BM-MSCs)相似的表型,且其能上调CD4~+CD25~+Foxp3~+T细胞的频率而产生免疫抑制功能。结论AF-MCs具有与BM-MSCs类似的表型,并且在免疫调节方面与BM-MSCs类似。这为临床应用间充质干细胞提供了一个新的,安全的种子细胞来源。     

High- and low-level cytokine induction in human peripheral blood mononuclear cells by different Borrelia burgdorferi strains

In this report we have compared the ability of 14 Borrelia burgdorferi sensu lato isolates to stimulate monocytes. From these isolates, all three human pathogen genospecies were represented. To determine the stimulatory activity of the different strains, interleukin-1β (IL-1β) was measured in the supernatant of monocyte cultures. This was achieved with borrelial strains in a ratio of 10 bacteria to 1 monocyte. In the majority of strains the stimulation induced a release of about 8000 pg/ml IL-1β, whereas four strains (B31, 297, EB3, 1/B29) induced more than 18000 pg/ml IL-1β. We could, therefore, define two groups: low-level inductors and high-level inductors for IL-1β. The strains in the defined groups could not be ascribed to one distinct genospecies or a biological source. Further experiments confirmed the same differential release for tumor necrosis factor-α, but not for IL-6. Studies on IL-1β indicated that high- and low-level release of cytokine was due to differences in protein synthesis. Received: 30 January 1996     

自外周血记忆B细胞体外活化和分化抗原特异性浆细胞的方法研究

目的 建立自人外周血淋巴细胞(PBMCs)中的记忆B细胞活化和分化特异性抗体分泌细胞(如浆细胞)的体外活化方法,为抗体药物研发提供研究基础.方法 采集两名接种过乙型肝炎疫苗3年和25年健康成年志愿者(Z和L)外周血,分离PBMCs,通过白细胞介素(IL)-2、IL-4和Toll样受体诱导剂(CpG、R848)体外诱导活化记忆B细胞.采用酶联免疫吸附试验(ELISA)检测培养上清中总抗体和HBsAg特异性抗体的含量;通过酶联免疫斑点试验(ELISPOT)检测培养细胞中总抗体分泌细胞(ASC)数量的变化和HBsAg特异性抗体细胞数量.结果 活化第6天,对照组和两个活化组[CpG DNA2006联合IL-2(C组)和R848联合IL-2(R组)]细胞培养上清总免疫球蛋白(Ig)G浓度分别为37.06ng/mL、80.87 ng/mL和85.97 ng/mL,两个活化组(C组和R组)均明显高于对照组(t=23.318,60.639,P均＜0.05).ELISPOT检测ASC数量,结果显示,C组和R组数量均多于对照组,并且R组明显多于C组.两种活化方法均能在体外诱导B细胞活化,但R组活化效果更佳,确立R组为最佳活化方法.用该方法活化两名乙型肝炎疫苗接种者PBMCs,ELISA检测活化组特异性抗体浓度,R组显著高于对照组(t=5.031,11.561,P均＜0.05).不同细胞数量诱导的分组,ELISPOT检测HBsAg特异性抗体分泌细胞数量,R组均明显多于对照组.结论 确立诱导剂R848和细胞因子IL-2联合能够有效地诱导外周血记忆B细胞体外活化和分化抗原特异性浆细胞,为今后抗体药物研发提供了研究基础.     

内皮细胞在感染性休克炎症性细胞因子释放中的作用

目的 以感染性休克患者血清刺激人原代内皮细胞(HPAEC)和外周血单个核细胞(PBMC),观察内皮细胞在感染性休克细胞因子释放中的作用.方法 采用密度梯度离心法分离健康人PBMC;RT-PCR及免疫印迹检测HPAEC细胞表面标志CD144和血管假性血友病因子(von Will-ebrand factor,vWF)的表达;ELISA检测感染性休克患者和健康对照者血清中的IL-6、TNF-α、MCP-1的水平;两种血清及LPS刺激HPAEC和PBMC后,ELISA检测培养上清中IL-6、TNF-α、MCP-1的水平;流式细胞技术检测休克患者血清和LPS刺激后HPAEC膜表面ICAM-1的表达;S1P1受体激动剂CYM-5442预处理后,休克血清刺激HPAEC培养上清中IL-6、TNF-α、MCP-1的水平.结果 HPAEC表达内皮细胞标志性分子CD144和vWF;感染性休克患者血清中炎症因子IL-6、TNF-α、MCP-1水平明显高于健康对照(P＜0.01);休克血清和LPS处理PBMC和HPAEC后,相对于对照组,均有明显升高(P＜0.01);PBMC的休克组炎症因子与PBMC的LPS组均无明显差异(P＞0.05),而HPAEC的休克组炎症因子相对于HPAEC的LPS组均明显升高(P＜0.01);同样地,LPS刺激两种细胞后,HPAEC的IL-6、TNF-α明显低于PBMC(P＜0.01),MCP-1的水平无明显变化(P＞0.05),但是休克血清刺激后,HPAEC的IL-6、TNF-α与PBMC无明显变化(P＞0.05),MCP-1则明显升高(P＜0.01);休克患者血清刺激S1P1受体特异性激动剂CYM-5442预处理的HPAEC后,培养上清中炎症因子IL-6、TNF-α、MCP-1水平明显降低(P＜0.01).结论 内皮细胞在感染性休克细胞因子释放中起着关键作用.     

乳腺癌外周血单个核细胞与间质免疫微环境的关系研究

目的探讨乳腺癌免疫微环境与外周血的关系。方法应用BRB-Array Tools软件对公共基因芯片数据库GEO中的乳腺癌间质及乳腺癌患者外周血单个核细胞基因芯片表达数据进行统计学分析,找出在乳腺癌间质及外周血单个核细胞均发生变化的基因,DAVID工具进一步分析其功能及参与的生物学通路。PINA蛋白质互作平台分析这些基因的蛋白质相互作用情况。结果比较后得到共同差异表达的103条基因,失调方向一致的基因70条,功能涉及炎症反应、髓系细胞分化、白细胞激活、抗原加工提呈等多种免疫相关的生物学过程。结论乳腺癌患者外周血单个核细胞基因表达改变,与肿瘤间质微环境具有一定相似度,有望建立基于外周血的免疫微环境分子预测,为乳腺癌的治疗靶点及预后判断的研究开辟新思路。     

Simvastatin induces interleukin-18 production in human peripheral blood mononuclear cells

The effects of statins on immune response depend on the inhibition of 3-hydroxy-3-methylglutaryl coenzyme-A (HMG-CoA) reductase and leukocyte function-associated antigen (LFA)-1, which is a ligand of intercellular adhesion molecule (ICAM)-1. Simvastatin, an HMG-CoA reductase inhibitor with mild inhibition of LFA-1, induced the production of interleukin (IL)-18, tumor necrosis factor (TNF)-alpha and interferon (IFN)-gamma in human peripheral blood mononuclear cells (PBMC). The IL-18 production is located upstream of the cytokine cascade activated by simvastatin. Moreover, simvastatin concentration-dependently inhibited the expression of ICAM-1 and induced the expression of CD40 on monocytes. In the presence of IL-18, simvastatin suppressed the expression of ICAM-1 and CD40 as well as the production of IL-12, TNF-alpha and IFN-gamma in PBMC, contributing to the anti-inflammatory effect of simvastatin. The effects of simvastatin were abolished by the addition of the product of the HMG-CoA reductase, mevalonate, indicating the involvement of HMG-CoA reductase in the action of simvastatin.     

Effect of glutamine on Th1 and Th2 cytokine responses of human peripheral blood mononuclear cells

Decreased glutamine concentrations are found in patients with catabolic stress and are related to susceptibility to infections. In this study, we evaluated the role of glutamine in Th1/Th2 cytokine responses. Peripheral blood mononuclear cells were stimulated with phytohemagglutinin (PHA), live attenuated bacillus Calmette-Guérin (BCG), or measles virus in the presence of different glutamine concentrations. We found that glutamine at an optimal concentration (0.6 mM) significantly enhanced PHA-stimulated lymphocyte proliferation as well as Th1 [interferon-gamma (IFN-gamma) and interleukin-2 (IL-2)] and Th2 cytokine (IL-4 and IL-10) production. In the absence of glutamine, BCG and measles virus elicited minimal lymphocyte proliferation, whereas BCG enhanced Th1 cytokine response and measles virus promoted Th2 cytokine response. Interestingly, addition of glutamine promoted the BCG-elicited Th1 cytokine response (IFN-gamma), but suppressed the measles-induced Th2 cytokine response (IL-10). These results suggest that appropriate glutamine levels may influence host responses to different antigens and microorganisms. Furthermore, predominately Th1, but not Th2, cytokine responses required the presence of optimal concentrations of glutamine.