重症肌无力患者血液T细胞亚群和T细胞功能

本文对一组重症肌无力病人血液T细胞亚群及外周血单个核细胞产生白细胞介素2的能力进行了测定。结果发现病人的OKT_8~+细胞减少,OKT_4~+/OKT_8~+比值增高,产生白细胞介素2能力降低,这些变化在全身型病人较眼肌型明显。本文结合文献讨论细胞免疫的变化在本病发病机理中的作用,并提出IL—2对本病进行免疫调节治疗的前景。     

雷公藤多甙对重症肌无力患者外周血T淋巴细胞亚群的影响

目的探讨雷公藤多甙（ＴⅡ）对重症肌无力（ＭＧ）的免疫调节机制。方法将５２例ＭＧ患者随机分为ＴⅡ治疗组及非ＴⅡ治疗组,观察ＴⅡ治疗组和非ＴⅡ治疗组治疗前后及４０例正常对照外周血Ｔ淋巴细胞亚群分布。结果ＴⅡ治疗组及非ＴⅡ治疗组治疗前外周血中ＣＤ３、ＣＤ８细胞较正常对照组明显降低,ＣＤ４与正常对照组比较无显著性差别,ＣＤ４／ＣＤ８比值较正常对照组显著升高;ＴⅡ治疗组治疗前后外周血中ＣＤ３无明显差别,ＣＤ４较治疗前明显降低,ＣＤ８明显增加,ＣＤ４／ＣＤ８比值则显著降低,而非ＴⅡ治疗组外周血ＣＤ３、ＣＤ４、ＣＤ８、ＣＤ４／ＣＤ８比值治疗前后则无显著性变化。结论ＴⅡ治疗ＭＧ的免疫抑制机制可能通过调节Ｔ淋巴细胞亚群分布而发挥作用。     

重症肌无力患者外周血T淋巴细胞亚群凋亡的研究

目的探讨外周血T淋巴细胞亚群凋亡在重症肌无力(MG)免疫发病机制中的作用及临床意义。方法采用流式细胞术结合免疫荧光抗体法测定了45例MG患者和40例正常对照组外周血CD4~ T淋巴细胞、CD8~ T淋巴细胞百分数及其凋亡状况,并对24例MG患者在应用糖皮质激素治疗后外周血T淋巴细胞亚群凋亡状况进行了观察。结果MG患者组外周血CD8~ T淋巴细胞百分率较正常对照组明显降低,CD4~ /CD8~ 比值较正常对照组明显升高：MG患者组外周血CD4~ T淋巴细胞凋亡程度较正常对照组明显降低；MG患者外周血CD4~ T淋巴细胞凋亡程度与MG类型、预后密切相关：应用糖皮质激素治疗后MG患者外周血CD4~ T淋巴细胞凋亡程度明显增强。结论MG患者存在T淋巴细胞亚群的分布异常及CD4~ T淋巴细胞凋亡功能障碍,其凋亡程度可能与MG类型、预后密切相关,糖皮质激素对MG患者外周血CD4~ T淋巴细胞凋亡缺陷具有纠正作用。     

重症肌无力患者周围血中CD4+T淋巴细胞亚群的研究

目的研究ＣＤ４＋Ｔ淋巴细胞的功能亚群在重症肌无力（ｍｙａｓｔｈｅｎｉａｇｒａｖｉｓ,ＭＧ）发病中的作用。方法采用免荧光双标记技术和流式细胞仪对３９例ＭＧ患者和１８例健康对照者周围血中ＣＤ４＋Ｔ淋巴细胞的两个亚群（ＣＤ４＋ＣＤ４５ＲＡ＋、ＣＤ４＋ＣＤ４５ＲＡ－）的百分率进行测定。结果ＣＤ４＋ＣＤ４５ＲＡ＋亚群的百分率在ＭＧ组和ＡＣｈＲＡｂ阳性组明显低于对照组和ＡＣｈＲＡｂ阴性组;而ＣＤ４＋ＣＤ４５ＲＡ－细胞的百分率在ＭＧ组和ＡＣｈＲＡｂ阳性组显著高于对照组和ＡＣｈＲＡｂ阴性组。结论表明ＣＤ４＋Ｔ淋巴细胞的两个功能不同亚群在ＭＧ患者体内发生了异常改变,与ＡＣｈＲＡｂ的产生有关。     

重症肌无力患者外周血CD+4 CDhigh25 T细胞及其动态观察

目的 研究重症肌无力(myasthenia gravis, MG)患者外周血CD+4 CDhigh25 T细胞的水平,以及各种治疗方法对其的影响.方法 应用四色流式细胞仪检测55例MG患者(治疗前)与33名健康对照外周血CD+4 CDhigh25 T细胞百分率,对其中26例MG患者进行了治疗前后的动态检测.结果 MG患者与健康对照外周血CD+4 CDhigh25 T细胞百分率分别为6.22%±3.37%与5.16%±1.87%,两者差异无统计学意义(P=0.061),而21例非手术治疗患者其外周血CD+4 CDhigh25 T细胞百分率的变化与病情评分的变化呈负相关(r=-0.563, P=0.008).结论 这种非手术治疗前后短期内的CD+4 CDhigh25 T细胞的变化情况可能与病情相关,但胸腺切除前阶段观察的情况有所不同.     

重症肌无力患者胸腺细胞经AChR刺激后T淋巴细胞亚群Bcl-2的表达

目的研究重症肌无力(myasthenia gravis,MG)患者胸腺细胞经乙酰胆碱受体(AChR)刺激后T淋巴细胞亚群Bcl-2的表达水平和临床意义。方法应用流式细胞仪测定11例MG患者胸腺T淋巴细胞亚群经AChR刺激后Bcl-2表达水平。结果经AChR刺激后,MG组增生胸腺CD4、Bcl-2双阳性细胞明显高于对照组(P<0.01),CD8、Bcl-2双阳性细胞与对照组相比差异无统计学意义(P>0.05)。结论MG患者增生胸腺中AChR特异性辅助T细胞上Bcl-2表达增高,Bcl-2表达升高可能和AChR的诱导有关。     

雷公藤多甙对重症肌无力患者外周血T淋巴细胞 …

目的 探讨雷公藤多甙（ＴⅡ）对重症肌无力（ＭＧ）的免疫调节机制,方法 将５２例ＭＧ患者随机分为ＴⅡ治疗组及非ＴⅡ治疗组,观察ＴⅡ治疗组和非ＴⅡ治疗组治疗前后及４０例正常对照外周血Ｔ淋巴细胞亚群分布。结果 ＴⅡ治疗组及非ＴⅡ治疗组治疗前外周血中ＣＤ３,ＣＤ８细胞较正常对照组明显降低,ＣＤ４与正常对照组比较我显著性差别,ＣＤ４／ＣＤ８比值较正常对照组显著升高,ＴⅡ治疗组治疗前后外周血中ＣＤ３无明显差别     

重症肌无力胸腺淋巴细胞亚群表型的变化

目的　研究重症肌无力 (myastheniagravis ,MG)患者胸腺的T、B淋巴细胞亚群表型 ,并分析与其外周血的相关性。方法　应用免疫荧光标记技术 ,经流式细胞仪分析 ,检测了 59例MG患者 ,包括 30例伴胸腺病变患者外周血的淋巴细胞亚群表型。另外 ,对 8例MG患者的胸腺和外周血经体外培养后进行检测。结果　 ( 1)MG患者外周血辅助性T细胞 (Th ,CD4 )异常增高 ,病程 6个月以内、伴胸腺增生者Th细胞均明显增多 ;( 2 )MG胸腺和外周血中经乙酰胆碱受体 (AChR)刺激后活化的Th细胞 (CD4 CD2 5 )、CD5-B细胞 (CD5- CD19 )明显增多 ,且外周血CD5-B细胞与自身血清乙酰胆碱受体抗体 (AChRAb)滴度显著相关 (P <0 .0 1) ,胸腺摘除术 (Tx)后 ,MG外周血CD4 CD2 5 、CD5- CD19 细胞均有所减少。结论　MG患者胸腺存在着异常的AChR特异应答性T、B淋巴细胞亚群表型 ,尤其以活化的Th细胞为著 ;CD5-B细胞的产生可能与MG密切相关。     

重症肌无力患者外周血单核细胞亚群的研究

本文用抗单核细胞亚群M_1、M_2、M_3的单克隆抗体检测了重症肌无力(MG)患者的外周血单核细胞亚群,并同时测定了患者血清乙酰胆碱受体(AchR)抗体。结果发现MG病人的M_1升高,M_3下降,在疾病缓解期又趋正常。作者推测单核细胞异常所导致的免疫调节障碍在MG发病中起重要作用。     

Influence of T cell specificity on the antibody response to the acetylcholine receptor

Lymph node T cells were obtained from rats immunized with purified acetylcholine receptor (AChR) or isolated receptor subunits (alpha, beta, gamma, or delta). The immune T cells were then analyzed for the ability to respond to native AChR or AChR subunit challenge in an in vitro proliferation assay, as well as to perform as helper T cells (TH) in an in vitro anti-AChR antibody response. Results indicated a significant degree of subunit-to-subunit crossreactivity at the T cell level. Moreover, helper function could be generated by stimulation of T cells immune to any of the AChR subunits, although alpha-immune TH cells appeared to provide a quantifiably increased level of specific antibody production by AChR-immune B cells in vitro when compared to antibody produced by the same B cells in conjunction when beta-, gamma-, or delta-immune TH cells.     

重症肌无力T细胞受体基因重排的检测及其临床意义

目的胸腺病变情况与重症肌无力患者的治疗及预后密切相关，为了能早期诊断胸腺瘤。方法用Ｓｏｕｔｈｅｒｎ杂交，分别以ＴＣＲ－α、β及γ为探针检测了一组重症肌力无力患者Ｔ细胞受体（ＴＣＲ）基因重排。结果６例患者ＤＮＡ经ＥｃｏＲＩ酶切，与ＴＣＲ－α、β探针杂交时有一条异常的重排带。这６例患者已４例经手术证实为胸腺肿瘤。结论ＴＣＲ基因重排可能为早期诊断胸腺瘤提供帮助。     

Human muscle acetylcholine receptor reactive T and B lymphocytes in the peripheral blood of patients with myasthenia growth

The prevalence of T and B cells reactive with the acetylcholine receptor (AChR) of human skeletal muscle was studied in 33 patients with myasthenia gravis (MG), 18 patients with other neurological diseases (OND) or autoimmune disorders (AD) and 27 age- and sex-matched healthy controls. T cell stimulation was estimated by enumerating cells secreting interferon (IFN)-γ and interleukin (IL)-2 in response to the AChR, whereas B cell reactivity was estimated by enumerating cells secreting IgG antibodies binding to the AChR. AChR-reactive T cells were increased in the peripheral blood of patients with MG as compared to patients with OND, AD and healthy individuals. Of the patients with MG, 29/33 (87.7%) had numbers of IFN-γ secreting cells higher than the mean ± 2 SD of the mean of controls as compared to 4/18 (22.2%) of patients with OND or AD and 2/27 (7.4%) of the controls. The mean value of the numbers of AChR-reactive T cells in the patients with MG was 19.6/10⁵ PBMC, corresponding to 1/5100 PBMC. Comparable results were obtained also for IL-2-secreting cells. Anti-AChR IgG antibody-secreting cells were detected in the blood of 30/33 (91%) of the patients with MG, 3/18 (16.7%) of the patients with OND or AD and 2/25 (8%) of the controls. The mean value of the antibody-secreting cells in MG was 11.7 cells/10⁶ PBMC corresponding to 1/70400 PBMC in the patients with MG, compared to a mean value of antibody-secreting cells in the patients with OND or AD of 0.33 and controls of 0.16 cells/10⁶ PBMC.     

流式细胞仪检测成年人脑脊液T淋巴细胞亚群百分比参考范围

目的初步建立脑脊液T淋巴细胞亚群正常参考范围。方法使用四色流式细胞仪检测49例无神经系统疾病人群脑脊液和血液的T淋巴细胞亚群(CD3+CD4+、CD3+CD8+、CD3+CD69+)百分比水平。结果 (1)正常脑脊液T淋巴细胞亚群构成以T辅助细胞(CD3+CD4+T细胞)为主。T辅助细胞平均占脑脊液T淋巴细胞总数的68.27%,参考范围为54.77%～81.77%。T抑制细胞(CD3+CD8+T细胞)平均占脑脊液T淋巴细胞总数的31.73%,参考范围为18.23%～45.23%。活化T细胞(CD3+CD69+T细胞)平均占脑脊液T淋巴细胞总数的2.76%,参考范围为0.29%～26.29%。脑脊液CD4/CD8比值平均为2.21,参考范围为1.16～4.21。脑脊液的T辅助细胞百分比、活化T细胞百分比和CD4/CD8比值高于血液,T抑制细胞百分比低于血液(P<0.05)。(2)脑脊液与血液各T淋巴细胞亚群在各年龄组间无统计学差异。(3)脑脊液和血液各T淋巴细胞亚群在男女之间亦无统计学差异。结论本文初步建立了脑脊液T淋巴细胞亚群百分比的参考范围;正常脑脊液T淋巴细胞中以T辅助细胞为主,且其亚群分布特点与血液T淋巴细胞亚群不同。     

Recombinant interleukin 2 (IL-2) promotes T cell line-mediated neuroautoimmune disease

Autoimmunity is dependent on a delicate balance of cellular interactions preventing activation of autoaggressive T cells. Possible side effects of therapeutically injected recombinant interleukins on latent or overt autoimmune disease are uncertain.Using a T cell transfer model of autoimmune central nervous system (CNS) disease, we investigated the in vivo effect of recombinant IL-2.We observed that recombinant IL-2 strongly promotes autoimmune disease.     

Influence of T cell specificity on the heterogeneity and disease-causing capability of antibody against the acetylcholine receptor

Adoptive secondary anti-acetylcholine receptor (AChR) antibody responses were examined in rats to evaluate the influence of helper T cell specificity on the nature and disease-causing potential of antibody produced. Mixtures of B cells reactive with the intact AChR plus T cells reactive with purified AChR subunits (alpha, beta, gamma, delta) were transferred and antigen-challenged in immunologically naive recipient rats; the serum anti-AChR antibody produced was assessed by radioimmunoassay for differences in titers and by isoelectric focusing for differences in clonal heterogeneity as a function of the subunit specificity of T cells transferred. In addition, rats receiving different sources of AChR or AChR subunit-reactive T cells were examined for AChR-dependent muscle dysfunction. The results indicated a clear reduction in anti-AChR antibody concentrations and clonal heterogeneity in recipient rats receiving T cells of specificities restricted to individual subunits. However, except for a clear relationship between serum anti-AChR antibody concentration and disease induction, no particular AChR subunit-reactive helper T cell specificity appeared to preferentially cause muscle dysfunction. We conclude that if such relationships exists, T cells with specificities more restricted than those described here will have to be used.     

眼肌型及全身型重症肌无力患者外周血T淋巴细胞Fas的表达

目的 探讨Fas介导的细胞凋亡与眼肌型(ocular myasthenia gravis,OMG)及全身型重症肌无力(generalized myasthenia gravis,GMG)发病的关系.方法 采用流式细胞技术检测4例OMG、13例GMG患者及13例健康对照组外周血淋巴细胞中CD4、CD8及Fas的表达.结果 OMG、GMG组与对照组外周血T淋巴细胞表面CD4、CD8分子表达的差异无统计学意义(P>0.05).GMG组与对照组外周血T淋巴细胞中Fas+细胞比例的差异有统计学意义(41.72%±8.73%、31.22%±13.00%,P:0.017).GMG组与对照组Fas表达增高者比例的差异有统计学意义(61.5%、15,4%,P=0.041).Fas表达增高的GMG患者病情较重.病程较长.胸腺瘤发生率较高.OMG与GMG组外周血T淋巴细胞中Fa8+、CD4+Fas+、CD8+Fas+细胞比例差异无统计学意义(P>0.05).结论 GMG患者外周血T淋巴细胞中Fas的表达升高,OMG与GMG患者外周血T淋巴细胞中Fas的表达无显著差异,二者可能同属一种系统性疾病.     

重症肌无力患者细胞免疫及免疫调节 与临床疗效关系的分析

目的探讨周围血中ＮＫ细胞、Ｔ细胞亚群、细胞膜白细胞介素－２受体（ｍＩＬ－２Ｒ）阳性细胞与重症肌无力（ＭＧ）发病及临床疗效的关系。方法用流式细胞仪计数３９例ＭＧ患者ＣＤ３、ＣＤ４、ＣＤ８、ＣＤ２５和ＣＤ５６抗体阳性细胞的百分率,应用许氏临床评分法对病情的严重程度和转归进行评定。结果ＭＧ患者的ｍＩＬ－２Ｒ（ＣＤ２５）阳性细胞无明显改变。ＣＤ３阳性细胞在肾上腺皮素激素治疗前、后无明显改变。ＮＫ细胞（ＣＤ５６）和ＣＤ４阳性细胞在治疗前均明显高于正常（Ｐ＜００１）,治疗２个月后均明显下降（Ｐ＜０．０１,０．０５）。ＣＤ８阳性细胞治疗前后明显降低和增高（Ｐ＜００５）。结论ｍＩＬ－２Ｒ阳性细胞百分率不能直接反映ＭＧ的免疫功能紊乱,并对ＮＫ细胞和Ｔ细胞亚群的变化无直接调节作用。ＮＫ细胞增高,可能为其功能下降的代偿性变化。ＣＤ４增高,ＣＤ８下降提示免疫功能紊乱,符合自身免疫性疾病的特点。     

Efficacy of intranasal administration of neostigmine in myasthenic patients

Summary The efficacy of intranasally administered neostigmine was tested in 22 patients with generalized myasthenia gravis (MG). Topical therapy to the highly vascularized oropharynx proved to be quickly effective in 5–15 min both clinically and electrophysiologically. Twenty-eight MG patients were then recruited from different centres and their morning doses of oral pyridostigmine were substituted with intranasal neostigmine over a period of 2 or 3 weeks. Intranasal neostigmine proved to be equally efficacious in this regimen. No side-effect was noted even in 4 patients treated in this way for 1 year. Intranasal administration of anti-acetylcholinesterase may be very beneficial: (1) for patients with irregular absorption of oral doses; (2) early in the morning and every time a fast and temporary effect is needed; (3) in bulbar impairment and emergencies, in which a handy atomizer may be life-saving.Presented in part at the XIV World Congress of Neurology, New Delhi, 22–27 October 1989