粘附分子在实验性变态反应性脑脊髓炎发生中的作用

目的 探讨粘附分子ＣＤ１１ａ，ＣＤ５４，ＣＤ６２在实验性变态反应性脑脊髓炎（ＥＡＥ）发生中的作用。方法 采用Ｗｉｓｔａｒ大鼠建立了ＥＡＥ动物模型，应用免疫荧光法检测下沉大鼠及ＥＡＥ动物周围血和脑组织中Ｔ细胞亚群及ＣＤ１１ａ，ＣＤ５４，ＣＤ６２的表达，结果 发病大鼠脑组织局灶周围大量淋巴细胞浸润，主要为ＣＤ４Ｔ细胞，发病大鼠周围血淋巴细胞上及大鼠脑组织粘附分子的表达明显升高，结论 粘附分子ＣＤ１１ａ     

EAN血清可溶性细胞间粘附分子—1的测定

目的探讨血清可溶性细胞间粘附分子－１（ｓＩＣＡＭ－１）在实验性变态反应性神经炎（ＥＡＮ）中的作用。方法用兔坐骨神经匀浆加完全福氏佐剂（ＣＦＡ）免疫大鼠，建立ＥＡＮ模型；同时用抗细胞间粘附分子－１（ＩＣＡＭ－１）单克隆抗体注入大鼠体内后再诱导ＥＡＮ；观察自然病程组、抗体注射组及对照组的发病情况与病理特点；应用双抗体夹心ＥＬＩＳＡ法检测不同发病程度ＥＡＮ动物血清中ｓＩＣＡＭ－１的浓度。结果抗体注射组发病率及发病程度明显低于自然病程组；自然病程组ｓＩＣＡＭ－１的浓度高于抗体注射组及对照组；ＥＡＮ发病程度与ｓＩＣＡＭ－１的浓度呈正比。结论ｓＩＣＡＭ－１与ＥＡＮ的发病关系密切，ｓＩＣＡＭ－１的测定是观测自身免疫性疾病的一个有用的指标；抗ＩＣＡＭ－１抗体能够减轻或预防ＥＡＮ的发生。     

脑缺血抗细胞间粘附分子—1抗体作用的实验研究

目的探讨抗细胞间粘附分子－１（ＩＣＡＭ－１）抗体保护神经元缺血性损伤的作用机制。方法分离培养鼠脑毛细血管内皮细胞（ＣＣＥＣ）和多形核白细胞（ＰＭＮ），利用微管吸吮技术，观察ＰＭＮ与ＣＣＥＣ间粘附力学特性的变化。结果脑缺血－再灌注后各时间点，ＰＭＮ与ＣＣＥＣ的粘附力和粘附应力均明显高于正常对照组和伪手术组（Ｐ＜０．０１）；加抗ＩＣＡＭ－１抗体后，细胞粘附力和粘附应力均明显下降（Ｐ＜０．０５或Ｐ＜０．０１）。结论脑缺血－再灌注损伤后抗ＩＣＡＭ－１抗体使ＰＭＮ与ＣＣＥＣ粘附力减小，粘附应力下降；抗粘附分子抗体将可能成为治疗缺血性脑血管疾病的一条新的有效途径     

急、慢性炎性脱鞘性多神经根神经病患者CD+4T细胞、CD+8T细胞及NK细胞活性的动态研究

目的ＣＤ＋４Ｔ细胞、ＣＤ－８细胞以及ＮＫ细胞杀伤活性在炎性脱鞘性神经病中的变化规律和意义。方法分离外周单核淋巴细胞,采用ＡＰＡＡＰ法研究Ｔ细胞亚群、ＬＤＨ释放试验检测ＮＫ细胞活性。结果。ＡＩＤＰ发病初期ＣＤ４＋Ｔ细胞为主．在疾病的恢复期以ＣＤ８－Ｔ细胞为主,ＮＫ细胞的活性早于Ｔ细胞亚群的改变。ＣＩＤＰ病人ＣＤ＋８Ｔ细胞远远超过ＣＤ－４Ｔ细胞,在整个临床疾病过程中ＮＫ细胞活性持续较低。结论,ＡＩＤＰ和ＣＩＤＰ存在着不同的免疫损伤机制在疾病的不同时期ＣＤ４＋Ｔ和ＣＤ８－Ｔ细胞发挥了不同的作用,ＮＫ细胞可能具有重要的调节作用。     

重组白细胞介素2及T细胞亚群对重症肌无力患者AChR—ab的影响

对１９例全身型重症肌无力（ＭＧ）患者周围血Ｂ细胞分化能力进行了研究，并观察重组白细胞介素２（ｒＩＬ－２）及Ｔ细胞亚群对其影响。方法分离外周血纯Ｂ细胞，体外与电鳗乙酰胆碱受体（ＡＣｈＲ）一起培养，并分别加入去ＣＤ４＋Ｔ细胞，去ＣＤ８＋Ｔ细胞及ｒＩＬ－２。用酶联免疫－生物素法检测其上清液的乙酰胆碱受体抗体（ＡＣｈＲ－ａｂ）。同时以１０例健康人为对照组。结果去ＣＤ８＋Ｔ细胞后ＡＣｈＲ－ａｂ的产生无明显增加；去ＣＤ４＋Ｔ细胞加入ｒＩＬ－２后ＡＣｈＲ－ａｂ产生减少。结论提示ｒＩＬ－２对ＭＧ患者有潜在治疗价值。     

Alzheimer病的T淋巴细胞功能活性检测

目的　探讨外周Ｔ淋巴细胞功能活性与Ａｌｚｈｅｉｍ ｅｒ病（ＡＤ）病情的关系。方法　流式细胞仪测定Ｔ细胞亚群;ＭＴＴ法检测ＣｏｎＡ刺激ＡＤ外周淋巴细胞的增殖功能;ＥＬＩＳＡ法检测细胞因子及其受体表达。结果　（１）ＣＤ４＋ 、ＣＤ８＋ Ｔ细胞比例和ＣＤ４＋ ／ＣＤ８＋ 比值各组间无统计学差异。（２）淋巴细胞增值在轻、重ＡＤ组略下降,但刺激指数（ＳＩ）都大于２,与对照无明显差异。（３）ＩＬ－２的分泌在重度ＡＤ组和ｓＩＬ－２Ｒ表达在轻、重度ＡＤ组明显升高（Ｐ＜ ０．０５）,ＩＬ－２Ｒ的表达各组间差异无显著性。结论　Ｔ淋巴细胞亚群变化与ＡＤ病情的联系不明显,细胞因子分泌功能与ＡＤ进展有关。     

急,慢性炎性脱鞘性多神经根神经病患者CD^＋4T细胞,CD^＋ …

目的 ＣＤ＾＋４Ｔ细胞,ＣＤ＾＋８细胞以及ＮＫ细胞杀伤活性在炎性脱鞘性神经病中的变化规律和意义。方法 分离外周单核淋巴细胞,采用ＡＰＡＡＰ法研究了Ｔ细胞亚群,ＬＤＨ释放试验检测ＮＫ细胞活性,结果 ＡＩＤＰ发病初期ＣＤ＾＋８Ｔ细胞为主,在疾病的恢复期以ＣＤ＾＋８Ｔ细胞为主,ＮＫ细胞的活性早于Ｔ细胞亚群的改变,ＣＩＤＰ病人ＣＤ＾＋８Ｔ细胞远远超过ＣＤ＾＋４Ｔ细胞,在整个临床疾病过程中ＮＫ细胞活性持续较     

实验性超负荷血糖大鼠脑缺血后脑毛细血管内皮细胞ICAM-1表达的研究

目的　了解超负荷血糖条件下，脑缺血后，脑毛细血管内皮细胞细胞间粘附分子１（ＩＣＡＭ－１）表达的情况。方法　采用ＳＤ大鼠尾静脉注射链脲霉素，建立超负荷血糖模型。用免疫组化方法动态观察大鼠超负荷血糖１ 月条件下以尼龙线栓堵大鼠大脑中动脉造成持续性局灶性脑缺血后不同时间，脑毛细血管内皮细胞ＩＣＡＭ－１ 的表达。结果　超负荷血糖１月脑缺血０．５ 小时ＩＣＡＭ－１ 的表达明显升高；１小时达高峰，表达范围弥漫整个缺血半球；缺血３～１２ 小时， ＩＣＡＭ－１表达仍很明显，但主要集中在颞、顶叶缺血区；缺血２４ 小时表达不明显；假手术组大鼠脑组织毛细血管内皮细胞ＩＣＡＭ－１只有较少的表达；正常血糖对照组及阴性对照者脑毛细血管内皮细胞均未见ＩＣＡＭ－１ 表达。结论　与正常血糖ＳＤ大鼠脑缺血相比，超负荷血糖大鼠脑缺血后， ＩＣＡＭ－１ 在脑毛细血管内皮细胞上表达出现的早而明显，提示对加重脑缺血后的病理损伤起到了重要作用。     

精神分裂症病人的外周血免疫学参数

为探讨不同亚型精神分型精神分裂症患者的免疫功能状态，对４１例病人Ｔ细胞亚群、ＮＫ细胞ＣＤ５６的的表达、白细胞介素４（ＩＬ－４）及可溶性白细胞介素２受体进行检测，并以１５例健康人为对照。结果显示，Ⅰ型患者ＣＤ３＋、ＣＤ４＋细胞数、ＣＤ４／ＣＤ８比值显著低于对照组，ＣＤ５６＋细胞数、ｓＩＬ－２Ｒ显著高于对照组；Ⅱ型患者ＣＤ８＋细胞数明显升高，ＣＤ４／ＣＤ８比值下降，而ＣＤ５６＋、ＣＤ３＋、ＣＤ４＋ＩＬ     

脑血管病患者细胞间粘附分子—1的测定及意义

目的：探讨脑血管病患者血清细胞间粘附分子－１（ＩＣＡＭ－１）水平和脑血管病发病与治疗的关系。方法：用ＥＬＩＳＡ法测定脑血管病患者和正常对照组血清可溶性细胞间粘附分子－１（ｓＩＣＡＭ－１）含量。结果：脑血栓形成或脑出血患者血清ｓＩＣＡＭ－１含量均较对照组显著增高（Ｐ〈０．０１;Ｐ〈０．０１）;Ⅲ伴有高血压的脑血栓形成、脑出血患者血清ｓＩＣＡＭ－１含量较正常血压者显著增高（Ｐ〈０．０５）。结论：ＩＣＡ     

GSNO attenuates EAE disease by S-nitrosylation-mediated modulation of endothelial-monocyte interactions

S-Nitrosoglutathione (GSNO) is an endogenous nitric oxide carrier and recently, has been documented for its anti-inflammatory effects in rat model of cerebral ischemia (Khan et al. (2005) J Cereb Blood Flow Metab 25:177-192). Here, we explored the neuroprotective effects mediated by GSNO in Lewis rat model of EAE and its mechanism of action using in vitro model of monocyte-endothelial cell interaction. Oral administration of GSNO attenuated the clinical disease course in EAE animals by inhibiting the infiltration of vascular immune cells in the CNS that subsequently led to the reduction in the expression of proinflammatory cytokines and consequently limited demyelination. Based on the inhibition in infiltration of immune cells, we hypothesized that GSNO modulated endothelial cell activation that led to reduce cellular infiltration in the CNS. Using an in vitro model, we established that GSNO inhibited monocyte adhesion to the activated endothelial cell, which was mediated by down regulation of endothelial cell adhesion molecules (CAMs). The mechanism by which GSNO modulated CAMs expression appeared to be via S-nitrosylation of p65, which consequently inhibited nuclear factor kappa B (NF-kappaB) activation in endothelial cells. These observations suggest that GSNO exerts its protective effects in EAE by inhibition of cellular infiltration into the CNS by S-nitrosylation of p65, thereby modulating NF-kappaB-CAMs pathway in endothelial cells.     

Serum and cerebrospinal fluid levels of soluble adhesion molecules in multiple sclerosis: predominant intrathecal release of vascular cell adhesion molecule-1

Activated cerebral vascular endothelial cells express leukocyte, vascular cell, and intracellular adhesion molecules (E-selectin, VCAM-1 and ICAM-1) which facilitate leukocyte adhesion to endothelium and migration into inflammatory lesions. Paired serum and cerebrospinal fluid (CSF) levels of soluble (s) E-selectin, sVCAM-1 and sICAM-1 were determined by ELISA in patients with clinically definite MS in relapse, and patients with other inflammatory (IND) and non-inflammatory neurological disease (NIND). CSF levels of sVCAM-1 and sICAM-1 were significantly increased in MS patients compared to IND and NIND patients. Elevation of CSF sVCAM-1 in MS patients was the most marked finding (P = 0.0001) and an increased sVCAM-1 index indicated that this was due to intrathecal release of sVCAM-1. There were no differences in serum and CSF sE-selectin levels between the study groups. Measurement of the sVCAM-1 index may provide a marker of disease activity in patients with clinically definite MS.     

NCAM1与致痫大鼠认知功能障碍发病机制之间相关性的研究

目的 研究神经细胞黏附分子1(NCAMl)与癫痫大鼠认知功能障碍发病机制之间的相关性,探讨其在癫痫认知功能障碍中发挥的作用. 方法 采用随机数字表法将120只Wistar大鼠分为实验组和对照组.实验组又分为致痫组、卡马西平治疗组、奥卡西平治疗组、茴拉西坦治疗组、盐酸多奈哌齐治疗组,每组20只；采用匹罗卡品诱导癫痫模型,后4组并灌服相应药物.对照组(n=20)不造模,灌服生理盐水造模.通过Morris水迷宫实验测试大鼠的学习记忆能力,并通过RT-PCR法检测大鼠海马组织中NCAM1 mRNA表达,免疫组化法检测大鼠海马组织中NCAM1蛋白表达. 结果 各组大鼠水迷宫实验逃逸潜伏期差异有统计学意义(F=91.920,P=0.000),按长短排序为:卡马西平组＞奥卡西平组＞茴拉西坦组＞致痫组＞盐酸多奈哌齐组＞对照组.各组大鼠免疫组化及RT-PCR结果差异亦有统计学意义(F=324.510,P=0.000; F=81.160,P=0.000),按表达量多少排序为:盐酸多奈哌齐组＞茴拉西坦组＞致痫组＞奥卡西平组＞卡马西平组＞对照组. 结论 癫痫发作30d后海马的NCAM1表达水平升高,参与了癫痫认知功能障碍的发病；抗癫痫药物能加重癫痫认知功能障碍的发生,促智药物可明显改善癫痫的认知功能.     

肺移植前后可溶性细胞间黏附分子1的变化

背景：应用生物素-链亲和素系统建立的可溶性细胞间黏附分子1测定法,是一种敏感度高、可测范围广的检测方法。 目的：建立可溶性细胞间黏附分子1生物素-链亲和素系统时间分辨荧光免疫分析方法并应用于临床,探讨肺移植前后血清可溶性细胞间黏附分子1的变化及临床意义。 方法：使用2株匹配的单克隆抗体分别作为捕获抗体和检测抗体,利用制备的铕标记链亲和素(SA-Eu3+)作为示踪物并与生物素化的检测抗体特异结合,建立双位点多层夹心法,通过对30例健康人血清可溶性细胞间黏附分子1检测和26例肺移植受者手术前后血清可溶性细胞间黏附分子1的变化测定,对可溶性细胞间黏附分子1生物素-链亲和素系统方法学和在肺移植术前后的临床应用价值进行评价。 结果与结论：30例健康对照测定结果为(348.63±69.12) µg/L。移植前可溶性细胞间黏附分子1与对照组无显著性差别;移植后,各组与对照组之间比较差异有显著性意义(P < 0.05),急性排斥反应时血清可溶性细胞间黏附分子1升高,并发感染时降低,但各组之间比较无明显差别。表明可溶性细胞间黏附分子1生物素-链亲和素系统是一种新型的高灵敏度、宽范围的非放射性标记免疫分析法,对肺移植受者移植前后监测血清可溶性细胞间黏附分子1可作为辅助诊断急性排斥反应的免疫学指标。     

Increased expression of adhesion molecule CD18 (LFA-1β) on the leukocytes of peripheral blood in patients with acute ischemic stroke

Objectives - The aim of this study was to investigate whether adhesion molecules play a role in acute ischemic stroke. Material and methods -Using immunofluorescence phenotyping and flow cytometry, the expression of leukocyte adhesion molecules CD54, CD11a, CD11b and CD18 in peripheral blood were measured within 12 h after onset of ischemia in 20 patients with stroke. Follow-up measurements were performed at 7 and 30 days after ictus. Results - CD 18 immunofluorescence was significantly increased on the leukocytes within 12 h after onset in patients with stroke compared with the age-matched control group (20 patients with other neurological diseases). Follow-up measurement of CD18 revealed normal results as found in the control group. Conclusion - Our data support the idea that adhesion molecules are involved in tissue injury in ischemic stroke.     

脑梗死血清可溶性内皮细胞白细胞粘附分子-1的变化△

目的　探讨急性脑梗死血清可溶性内皮细胞白细胞粘附分子－１（ｓＥＬＡＭ－１）的变化及其临床意义。方法　采用ＥＬＩＳＡ测定了６９ 例脑梗死患者血清ｓＥＬＡＭ－１的变化,并与２３ 例脑出血和２０ 例正常人对照比较。结果　脑梗死患者２４ ｈ 内血清ｓＥＬＡＭ－１ 水平明显高于脑出血和正常对照组（Ｐ＜ ０．０１）。血清ｓＥＬＡＭ－１ 水平在２４ｈ 至１４ 天（ｄ）呈下降趋势。大梗死灶组血清ｓＥＬＡＭ－１ 水平明显高于中、小梗死灶组。脑梗死后并发感染者血清ｓＥＬＡＭ－１ 水平明显升高。结论　ｓＥＬＡＭ－１ 与急性脑梗死密切相关,深入研究ｓＥＬＡＭ－１ 在脑梗死中的作用有重要的临床应用价值     

Up-regulation of intercellular adhesion molecule 1 in cerebral microvessels after cortical contusion trauma in a rat model

A study was made on the expression of the intercellular adhesion molecule 1 (ICAM-1) in cerebral microvessels after cortical contusion trauma of the rat brain. The trauma was produced by a free-falling weight on the exposed dura of one fronto-parietal lobe. Immunohistochemistry was done on cryostat sections using a monoclonal antibody and the reaction product was visualized using the avidin-biotin-peroxidase complex method. Control and sham-operated rats showed immunostaining of some penetrating arteries of the cerebral cortex, the epithelial cells of the choroid plexus and occasional microvessels of the brain parenchyma. The same pattern of immunostaining was seen in rats that were subjected to trauma and killed after 30 min. All rats with contusion trauma that were allowed to survive for 6–72 h showed a substantial increase in the number of immunostained capillaries throughout the site of the lesion. The ipsilateral hippocampus showed a mild to moderate increase in the number of immunostained microvascular profiles. This phenomenon was also present in the lateral thalamus of some rats. The staining was seen as an uninterrupted line at the position of the endothelial cells, indicating an up-regulation of this adhesion molecule after brain trauma. Up-regulation of ICAM-1 is a well-known phenomenon in inflammatory and ischemic lesions of the brain but has not previously been described in detail in traumatic brain injury. ICAM-1 may be involved in the production of several post-traumatic events such as leukocyte adhesion, microcirculatory disturbances and edema formation. Received: 23 September 1996 / Revised, accepted: 18 November 1996     

The cytoplasmic domain of NrCAM binds to PDZ domains of synapse-associated proteins SAP90/PSD95 and SAP97

NrCAM, a member of the L1 family of cell adhesion molecules, serves important functions during the development of the nervous system, e.g. in adhesion-dependent processes such as neurite outgrowth and axonal pathfinding. Complex homo- and heterophilic binding and several extracellular ligands of NrCAM have been described, but less is known about intracellular interaction partners. The cytoplasmic carboxy-terminus of NrCAM contains a typical sequence motif for binding to PDZ domains, making interactions with PDZ domain-containing scaffolding proteins quite conceivable. In this study, we identified specific interactions of the intracellular domain of NrCAM with class I PDZ domains of the membrane-associated guanylate kinases SAP90/PSD95 and SAP97. In contrast to NrCAM, the intracellular domains of the other mammalian L1 family molecules, e.g. L1, CHL1 and Neurofascin, did not interact with these PDZ domains. In transfected COS-7 cells, NrCAM-mediated recruitment of SAP97 to the plasma membrane was dependent on the PDZ binding motif. We show that NrCAM and SAP97 are colocalized, e.g. within photoreceptor terminals of the mammalian retina. In summary, our results confirm a functional PDZ domain binding motif at the carboxy-terminus of NrCAM and support potential functions of NrCAM during the assembly of highly organized protein complexes at the cell membrane.