细胞因子与重症肌无力

重症肌无力（ＭＧ）是由乙酰胆碱受体抗体（ＡｃｈＲａｂ）介导的具有细胞免疫（ＧＭＩ）依赖性补体参与的,针对神经肌接头突触后膜上乙酰胆碱受体（ＡｃｈＲ）的自身免疫性疾病（ＡＩＤ）,是器官特异性ＡＩＤ的原型。曾认为体液免疫异常是ＭＧ唯一致病因素,但１０％～...     

重症肌无力患者胸腺组织CMTM8、CKLF1及相关趋化因子的表达

目的:比较重症肌无力(MG)患者与非自身免疫对照组胸腺组织CMTM8、CKLF1、相关趋化因子及受体的表达,为揭示MG胸腺异常及其发生机制提供理论依据.方法:用基因芯片筛选差异表达的趋化因子基因,采用RT-PCR和实时荧光定量PCR方法分别检测正常胸腺和MG胸腺中CMTM8 mRNA的表达,用免疫组织化学方法对CKLF1蛋白的表达和分布进行检测.结果:增生型MG 胸腺组织中CMTM8 mRNA的表达明显高于正常人胸腺(2-△△Ct=8.3),差异有统计学意义;CKLF1蛋白在正常胸腺、MG胸腺中的均有广泛的表达,阳性细胞主要为胸腺基质细胞;CXCR6、CCL2、CCL8、 CX3CL1、CXCL10和CCR1在 MG患者胸腺表达水平显著低于对照组;CCL22、CCL3、CXCL12、CCR5、CCR7和CCR9基因在MG患者胸腺表达水平显著高于对照组.结论:CMTM8及多种趋化因子基因在增生型MG胸腺的表达明显高于对照组胸腺,可能在MG患者T细胞异常发育的某些阶段起重要作用.     

胸腺摘除对重症肌无力患者T细胞亚群变化的影响

采用免疫荧光染色技术，经流式细胞仪分析，观察了重症肌无力（ＭＧ）患者胸腺摘除前、后外周血Ｔ细胞亚群变化，并研究了淋巴细胞经植物血凝素（ＰＨＡ）激活后Ｔ淋巴细胞亚群的变化。结果显示：１．未经治疗的患者存在明显Ｔ细胞亚群异常，表现为ＣＤ３~＋、ＣＤ４~＋ＣＤ８~－和ＣＤ２９~＋ＣＤ４~＋百分率高，ＣＤ４~－ＣＤ８~＋和ＣＤ１６， ５６~＋百分率低下；２．胸腺摘除后，ＣＤ３~＋、ＣＤ４~＋ＣＤ８~－、ＣＤ２９~＋ＣＤ４~＋百分率降低，ＣＤ４~－ＣＤ８~＋百分率增高；３．ＰＨＡ对外周血中ＣＤ３~＋和ＣＤ２９~＋ＣＤ４~＋有明显下调作用；ＣＤ４~＋ＣＤ８~＋亚群在２４小时呈现上调，４８－７２小时呈现下调；４．ＰＨＡ作用７２小时，正常人ＣＤ４~＋ＣＤ８~－亚群无明显变化，对ＣＤ４~－ＣＤ８~＋亚群呈下调作用；对摘除胸腺的患者ＣＤ４~＋ＣＤ８~－亚群呈现明显下调，对ＣＤ４~－ＣＤ８~＋亚群则呈轻度下调作用。本研究结果表明：摘除胸腺后，ＣＤ４~＋ＣＤ８~－亚群减少，ＣＤ４~－ＣＤ８~＋亚群占优势。     

重症肌无力病人胸腺和血液淋巴细胞亚群测定

本文采用武系抗淋巴细胞单克隆抗体，用免疫荧光法，检测了３２例重症肌无力病人血液及２５例ＭＧ病人胸腺组织中的淋巴细胞亚群。结果：①病人胸腺ＷｕＤＲ＋．ＷｕＢ＋细胞增多，尤以增生性和正常胸腺明显；胸腺ＷｕＴ６＋细胞减少。②病人血液中ＷｕＤＲ＋．ＷｕＢ＋细胞数升高；ＷｕＴ８＋细胞数降低；两者变化以具有萎缩性胸腺最为显著。     

胸腺切除术治疗重症肌无力的细胞免疫学机制

胸腺切除术是治疗重症肌无力的主要方法之一.75%的重症肌无力患者伴有胸腺异常,增生胸腺存在AChR特异反应性T、B细胞,胸腺瘤中存在着T细胞的分化,有乙酰胆碱受体抗体(AChRAb)滴度升高.Tx可能去除了胸腺中导致B细胞活化的TH细胞,其相关的细胞因子也逐步下调,AChRAb逐步减少,最终MG的症状缓解.     

胸腺病理在重症肌无力胸腺切除手术后远期预后生存分析中的作用

目的：探讨重症肌无力胸腺切除术后影响远期生态的因素。方法：采用胸腺切除手术治疗170例重症肌无力患者,对其中124例进行了超过40个月的远期随访,运用COX回归模型分析影响远期预后的有关因素（包括胸腺的各病理类型、性别、年龄、病程、术前临床Osserman分型和治疗）。同时运用免疫组织化学染色检测36例胞腺石腊标本中的T细胞和B细胞的数量,并与远期预后做等级相关分析。结果：重症肌无力胸腺切除术后不同的胸腺病理类型远期生态率有明显差异,表现为胸腺增生＞良性胸腺瘤＞胸腺萎缩＞恶性胸腺瘤（P＜0.050,胸腺中T细胞和B细胞的数量与术后的远期预后呈显著负相关（P＜0.05）。结论：胞腺不同的病理类型是影响重症肌无力胸腺切除术远期预后的重要因素,而且胸腺中T细胞和B细胞的数量也与预后相关。     

重症肌无力患者骨髓、胸腺及外周血中IFN-γ、IL-4分泌细胞的检测

<正>重症肌无力（MG）是神经肌肉传递障碍的自身免疫性疾病,它是以抗乙酰胆碱受体（AchR）抗体为其主要的致病性抗体。目前认为该抗体的产生依赖于T淋巴细胞,T细胞所分泌的细胞因子参与了实验性自身免疫性重症肌无力（EAMG）的诱导和发病过程。干扰素-γ（IFN-γ）和白细胞介素-4（IL-4）分别由Th1和 Th2亚型细胞所分泌,在机体免疫调节中起重要作用。本实验检测了早期发病的MG手术病人的骨髓、胸腺及外周血中IFN-γ和IL-4分泌细胞数,以期了解是否Th1及Th2均参与了MG的发病过程。     

树突状细胞亚群与重症肌无力

重症肌无力(MG)是一种依赖T细胞的抗体介导的以神经肌肉接头处传递障碍为特征的自身免疫性疾病,与机体免疫调节失衡有关.树突状细胞(DC)是目前功能最强的抗原递呈细胞(APC),在调节机体免疫反应中起着重要作用.不同表型的DC在自身免疫性疾病中的作用不同,它们通过不同的途径参与MG的发病.DC分泌的细胞凶子在MG发病机制巾也起着重要作用.细胞因子干预诱导的耐受性DC在治疗MG中取得了巨大进步,有望成为治疗MG的一种新方法.     

重症肌无力患者胸腺内microRNA-29表达水平的探究

目的 检测重症肌无力(myasthenia gravis,MG)患者胸腺组织中microRNA-29的表达水平并探讨其与MG发病的相关性.方法 22例经手术治疗的MG患者胸腺组织作为实验组,22例经手术治疗的非MG伴胸腺异常患者胸腺组织作为对照组.通过实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)的方法检测实验组及对照组胸腺组织中microRNA-29表达水平,采用秩和检验(Mann-Whitney U test)分析实验组与对照组中microRNA-29表达水平.采用Spearman秩相关分析实验组胸腺中microRNA-29表达水平与定量重症肌无力(quantitative myasthenia gravis score,QMG)之间的相关性.结果 MG患者胸腺组织中microRNA-29表达水平较对照组显著增高(P=0.032);依据MG患者性别、年龄、临床特点、胸腺病理结果将实验组分成不同临床亚组,microRNA-29在各组间表达差异无统计学意义(P值分别为0.614、0.471、0.267、0.329);microRNA-29表达水平与QMG呈显著正相关性(r=0.689,P=0.000a).结论 microRNA-29在MG患者胸腺组织中表达增高,其表达水平与肌无力严重程度呈正相关,而与患者性别、年龄、临床类型、胸腺病理结果无明显关联.     

细胞因子在重症肌无力发病机制中的研究进展

重症肌无力(MG)主要是由乙酰胆碱受体抗体(AChR-Ab)介导,依赖细胞免疫和补体参与的使神经肌肉接头处突触后膜上乙酰胆碱受体受损的自身免疫性疾病.近年来的研究表明,细胞因子间的平衡紊乱与MG的发病有重要关系.因而深入了解不同细胞因子在MG中的作用可以更加明确MG的发病机制及对其治疗提出新的方案.     

CD45RA／CD45RO临床研究进展

目的：探讨CD45RA／CD45RO的研究现状从而有效的指导临床工作。方法：查阅近年来国内外CD45RA／CIM5RO的相关文献,对其最新研究进行总结。结果：作为跨膜酪氨酸磷酸蛋白酶,CIM5RA／CD45RO在哺乳动物淋巴细胞信号传导、增殖和活化中有重要作用,是区分初始和记忆T细胞的重要标志。结论：CD45RA／CD45RO的检测在人类自身免疫疾病和病毒感染性疾病的诊断方面有重要意义。     

CD45RA and CD45RO isoforms in infected malnourished and infected well-nourished children.

The aim of this study was to determine if the distribution in vivo of CD4(+)CD45RA(+)/CD45RO(-) (naive), CD4(+)CD45RA(+)/CD45RO(+) (Ddull) and CD4(+)CD45RO(+) (memory) lymphocytes differs in malnourished infected and well-nourished infected children. The expression of CD45RA (naive) and CD45RO (memory) antigens on CD4(+) lymphocytes was analysed by flow cytometry in a prospectively followed cohort of 15 malnourished infected, 12 well-nourished infected and 10 well-nourished uninfected children. Malnourished infected children showed higher fractions of Ddull cells (11.4 +/- 0.7%) and lower fractions of memory cells (20.3 +/- 1.7%) than the well-nourished infected group (8.8 +/- 0.8 and 28.1 +/- 1.8%, respectively). Well-nourished infected children showed increased percentages of memory cells, an expected response to infection. Impairment of the transition switch to the CD45 isoforms in malnourished children may explain these findings, and may be one of the mechanisms involved in immunodeficiency in these children.     

慢性乙型肝炎患者外周血CD45RA+、CD45RO+T淋巴细胞亚群的特点及临床意义

目的：探讨慢性乙型肝炎患者外周血CD4^＋CD45RA^＋、CD4^＋CD45RO^＋、CD8^＋CD45RA^＋和CD8^＋CD45RO^＋T淋巴细胞亚群的特点及其与肝病病情的关系。方法：采集46例轻中度慢性乙型肝炎患者、58例重度慢性乙型肝炎患者和30例健康人的外周抗凝血，应用流式细胞技术三色荧光分析法对其外周血中CD4^＋CD45RA^＋、CD4^＋CD45RO^＋、CD8^＋CD45RA^＋和CD8^＋CD45RO＋T淋巴细胞亚群进行检测。结果：轻中、重度慢性乙型肝炎患者与正常人相比，其外周血中CD4^＋、CD8^＋T细胞均无明显改变；CD8^＋CD45RA^＋T细胞均明显降低，CD8^＋CD45RO^＋T细胞均明显增高，而CD4^＋CD45RA^＋、CD4^＋CD45RO^＋T细胞均无明显改变；重度慢性乙型肝炎患者与轻中度慢性乙型肝炎患者相比，CD8^＋CD45RA^＋T细胞明显降低（P〈0.05），CD8^＋CD45RO^＋T细胞明显升高（P〈0.05）。结论：乙型肝炎慢性化过程中，CD8^＋CD45RO^＋T细胞起重要作用且与慢性乙型肝炎患者病情的进展呈正相关；检测CD4^＋CD45RA^＋、CD4^＋CD45RO^＋、CD8^＋CD45RA^＋和CD8^＋CD45RO^＋T淋巴细胞亚群比检测CD4^＋和CD8^＋T细胞亚群更能正确、充分、全面地了解慢性乙型肝炎的发病机制和预后，从而有效地指导临床治疗。     

Comparison of lymphokine secretion and mRNA expression in the CD45RA+ and CD45RO+ subsets of human peripheral blood CD4+ and CD8+ lymphocytes

Flow cytometric analysis of human peripheral blood T lymphocytes demonstrated that the majority of the CD4⁺ cells were CD29⁺ or CD45RO⁺ “mature” cells while the CD8⁺ cells were primarily CD45RA⁺ “naive” cells. After an initial separation into CD4⁺ and CD8⁺ cells and a secondary separation into CD45 subsets, lymphokine secretion was assessed after phorbol 12-myristate 13-acetate and ionomycin or fixed anti-CD3 stimulation. Within the respective CD45 subsets, CD4⁺ cells produced more interleukin (IL)-2, IL-4, and IL-6; but the CD8⁺ cells secreted more interferon-γ and granulocyte/macrophage-colony-stimulating factor. Tumor necrosis factor-α secretion was similar in the matched CD45 subsets. Northern analysis revealed a parallel pattern of lymphokine mRNA expression in the four lymphocyte subsets. These results suggest that human CD8⁺ peripheral blood lymphocytes have a significant capacity to secrete lymphokines, and that the low lymphokine production observed in unseparated CD8⁺ cells reflects the higher percentage of less functional CD45RA⁺ cells.     

普通变异型免疫缺陷病CD45RA^＋和CD45RO^＋T细胞亚群异常

用不同的单抗可将Ｔ淋巴细胞区分为不同的功能亚群：“处女”Ｔ细胞（ｎａｉｖｅ，ＣＤ４５ＲＡ￣＋）及“记忆”Ｔ细胞（ｍｅｍｏｒｙ，ＣＤ４５ＲＯ￣＋）。以直接双标记免疫荧光经流式细胞仪观察了普通变异型免疫缺陷病ＣＤ４￣＋及ＣＤ８￣＋细胞中ＣＤ４５ＲＡ￣＋　和ＣＤ４５ＲＯ￣＋细胞亚群的分布及变化，发现ＣＤ４／ＣＤ８比值倒置的患者ＣＤ４￣＋ＣＤ４５ＲＡ￣＋Ｔ细胞亚群显著减少，而ＣＤ８￣＋ＣＤ４５ＲＯ￣＋和ＣＤ８￣＋ＣＤ４５ＲＡ￣＋亚群明显增高。     

蕈样肉芽肿患者外周血单一核细胞CD45RA及CD45RO表达的研究

目的探讨蕈样肉芽肿（Mycosis fungoides,MF）患者外周血单个核细胞CD45RA及CD45RO的表达及其与MF发病的关系。方法应用双荧光抗体标记、流式细胞仪检测15例MF患者外周血单个核细胞CD45RA及CD45RO的表达。结果（1）MF患者外周血CD3^＋、CD3^＋CD8^＋细胞与正常对照比较差异不显著（P〉0.05）。（2）MF患者外周血CD4^＋细胞低于正常对照,差异非常显著（P〈0.001）。（3）MF患者外周血T细胞CD3^＋CD4^＋/CD3^＋CD8^＋比值低于正常对照,差异显著（P〈0.001）。（4）MF患者外周血CD45RA^＋细胞低于正常对照,差异非常显著（P〈0.001）,CD45RO^＋细胞高于正常对照,差异非常显著（P〈0.001）。（5）MF患者外周血CD45RO^＋/CD45RA^＋比值高于正常对照,差异非常显著（P〈0.001）。（6）MF患者外周血CD4^＋CD45RA^＋细胞低于正常对照,差异非常显著（P〈0.001）。（7）MF患者外周血CD4^＋CD45RO^＋细胞及CD8^＋CD45RO^＋细胞均高于正常对照,差异非常显著（均P〈0.001）。（8）MF患者外周血CD4^＋CD45RO^＋/CD4^＋CD45RA^＋比值及CD8^＋CD45RO^＋/CD8^＋CD45RA^＋比值均高于正常对照,差异非常显著（P〈0.01及P〈0.001）。结论MF患者外周血中,不仅存在CD4^＋亚群失调和CD4^＋/CD8^＋比值降低,而且在CD4^＋和CD8^＋亚群中也存在CD45RA^＋、CD45RO^＋亚群失调和CD45RO^＋/CD45RA^＋比值升高,从而导致的机体免疫功能紊乱,可能与MF的发病或病情加剧有关。     

慢性乙型病毒性肝炎患者外周血T淋巴细胞CD27和CD45RA表达的研究

目的分析慢性乙型病毒性肝炎患者外周血T淋巴细胞CD27和CD45RA的表达。方法采集分离健康人和慢性乙型病毒性肝炎患者外周血单个核细胞(PBMC),利用多种荧光标记抗体标记细胞表面分子,再用流式细胞仪检测CD8+T淋巴细胞表面CD27和CD45RA表达情况。结果31例慢性乙型病毒性肝炎患者CD8+CD45RA+CD27+T细胞占CD8+T细胞(29.03±13.18)%,低于28例健康对照组的(60.85±14.36)%,P<0.01。而CD8+CD45RA-CD27+T细胞占CD8+T细胞(30.31±24.11)%,显著高于健康对照组的(10.32±5.24)%,P<0.05。慢性乙型病毒性肝炎患者CD4+CD45RA+CD27+T细胞21.12±9.64%低于健康对照组的(60.89±17.93)%,P<0.01,而CD4+CD45RA-CD27+T细胞(54.28±18.75)%显著高于健康对照组的(27.16±9.24)%,P<0.01。结论健康人外周血CD8+和CD4+T淋巴细胞均以CD45RA+CD27+初始细胞表型为主,而慢性乙型病毒性肝炎患者外周血初始细胞减少,CD45RA-CD27+表型的T淋巴细胞明显增加。     

Expression in vivo of CD45RA,CD45RB and CD44 on T cell receptor-transgenic CD8+ T cells following immunization

We used mice transgenic for a major histocompatibility complex class I-restricted T cell receptor to study the changes of phenotype in vivo which follow priming by antigen of CD8 T cells. We show that following priming with peptide, CD44 on CD8 T cells is up-regulated. The change of phenotype was relatively stable, as primed CD8 cells isolated from thymectomized mice 6 weeks after priming still expressed increased levels of CD44. CD8 T cells in these mice are still responsive to peptide and could represent long-lived primed cells. No downregulation in vivo of the CD45RA or CD45RB isoforms was found, indicating that there is a differential regulation of the expression of CD44 and CD45RB by activated CD8 transgenic T cells. These results contradict earlier studies in vitro which showed that CD8 T cells which have been primed earlier belong to the CD45RA^? or CD45RB^? subset.     

Hyper-reactivity of mouse CD45RA − T cells

Mouse CD4 T cells have been partitioned into CD45RA and CD45RA^? subpopulations by means ot the monoclonal antibody 14.8. The CD45RA^? subpopulation proliferated more actively and generated more interleukin-4 (IL-4) in response to stimulation with anti-CD3 antibody and phytohemaglutinin, and more IL-2 in response to anti-CD3. This subpopulation is therefore hyper-reactive to these polyclonal stimulators, but does not show the bias towards T helper type 2 activity that has been found in studies with other related CD45 isoforms. No evidence of suppression was obtained by comparing proliferation of CD45RA^? cells in the presence and absence of CD45RA cells. Thus mouse CD4 T cells behave in these respects similarly to those of man, as is evident in a brief review of the quiescence-activation-quiescence cycle in the two species.     

Selective deficiency of CD4+/CD45RA+ lymphocytes in patients with ataxia-telangiectasia

Several immunological abnormalities have been observed in ataxia-telangiectasia (AT), the most consistent being defects of immunoglobulin isotypes, decreased T-cell numbers, and reduced proliferative responses to mitogens. We examined the distribution of T lymphocytes expressing distinctive surface Ag characteristic of naive (CD45RA+) and memory (CD29+, CD45RO+) T cells, in both CD4+ and CD8+ (bright and dim) lymphocytes from 13 AT patients, compared with healthy agematched controls. We found that, irrespective of age, patients with AT had a severe deficiency of CD4+/CD45RA+ lymphocytes. This decrease accounted for the reduction of total CD4+ cells, since the absolute numbers ofmemory CD4+ cells were not significantly different in AT and in controls. Functional tests revealed poor proliferative responses to phytohemagglutinin and normal responses to soluble Ag (tetanus toxoid) in AT patients. These data fit with the distribution ofnaive andmemory cells, which are known to respond predominantly to mitogens or to recall Ag, respectively. CD45RA molecules were normally expressed on CD8+ lymphocytes. This rules out a generalized defect of regulation or differential splicing as the cause of defective expression of CD45RA on CD4+ cells. The selective deficiency of CD4+CD45RA+ may provide a cellular basis for some functional T-cell abnormalities of AT patients. Furthermore, it might practically serve for an early, or even prenatal, diagnosis of this disease.