Lombardi V R, García M, Rey L, Cacabelos R
Biotechnology Division, EUROESPES, Basic and Clinical Neurosciences Research Center, Bergondo La Coruña, Spain.
J Neuroimmunol. 1999 Jun 1;97(1-2):163-71. doi: 10.1016/s0165-5728(99)00046-6.
In order to investigate the possibility of whether or not the lymphocytes of patients with Alzheimer's Disease (AD) are in an activated state, blood mononuclear cells from 45 AD patients and 45 healthy age matched controls were immunophenotyped by measuring the expression of CD3, CD4, CD7, CD8, CD25, CD28, CD56 and HLA-DR by flow cytometry. Circulating and in-vitro-produced cytokines were also measured by ELISA tests. CD7 and CD8 were significantly decreased in AD patients (48.3% and 18.2%, respectively) when compared to healthy subjects (63.2% and 28.3%, respectively). A significant increase in the CD4, CD25 and CD28 antigen expression was also observed in the AD group (55.3% 24.8% and 65.1%) with respect to healthy subjects (44.5%, 10.3% and 54.3%). In addition there was a significant difference in the extent of apoptosis in lymphocyte culture, as measured by mean fluorescence intensity (MFI) of Fas antigen (CD95) expression on CD4+ T cells in 6 AD patients (MFI = 36% and 43%, by anti-CD3 and hyperthermia mediated-apoptosis, respectively) with respect to 6 healthy individuals (MFI = 24% and 31%, by anti-CD3 and hyperthermia mediated-apoptosis, respectively), as well as in T-cell proliferation assay. A decline of Fas antigen expression on CD8+ subset was observed in the AD group with both stimuli (19% and 28%) comparing to the control group (29% and 39%). No differences were observed on circulating cytokines and spontaneous in vitro production of proinflammatory interleukin 1beta (IL-1beta), Tumor Necrosis Factor-alpha (TNF-alpha), IL-6 and IL-10 cytokines. Lipopolysaccharide (LPS)-stimulated in vitro production of IL-1beta, TNF-alpha, IL-6 and IL-10 measured by a whole blood culture system was significantly higher in AD patients comparing to controls. Furthermore, the observed differences were more evident at late stages of disease. These findings suggest that immunological tests, based on lymphocyte immunophenotyping combined with pro-inflammatory cytokine determinations and measurement of apoptosis in peripheral blood might represent a useful tool to obtain more insight into the pathogenesis of AD and into the level of immune activation which could characterize the pathological state of lymphocytes from individual AD patients.
为了研究阿尔茨海默病（AD）患者的淋巴细胞是否处于激活状态，通过流式细胞术检测45例AD患者和45例年龄匹配的健康对照者的血液单核细胞中CD3、CD4、CD7、CD8、CD25、CD28、CD56和HLA - DR的表达，进行免疫表型分析。还通过ELISA试验检测循环和体外产生的细胞因子。与健康受试者（分别为63.2%和28.3%）相比，AD患者的CD7和CD8显著降低（分别为48.3%和18.2%）。与健康受试者（44.5%、10.3%和54.3%）相比，AD组中CD4、CD25和CD28抗原表达也显著增加（分别为55.3%、24.8%和65.1%）。此外，通过6例AD患者CD4 + T细胞上Fas抗原（CD95）表达的平均荧光强度（MFI）测量淋巴细胞培养中的凋亡程度存在显著差异（通过抗CD3和热介导凋亡，MFI分别为36%和43%），而6例健康个体（通过抗CD3和热介导凋亡，MFI分别为24%和31%）以及在T细胞增殖试验中也是如此。与对照组（分别为29%和39%）相比，AD组在两种刺激下CD8 +亚群上Fas抗原表达均下降（分别为19%和28%）。在循环细胞因子以及促炎白细胞介素1β（IL - 1β）、肿瘤坏死因子 - α（TNF - α）、IL - 6和IL - 10细胞因子的体外自发产生方面未观察到差异。通过全血培养系统测量，脂多糖（LPS）刺激下AD患者体外产生的IL - 1β、TNF - α、IL - 6和IL - 10显著高于对照组。此外，观察到的差异在疾病晚期更为明显。这些发现表明，基于淋巴细胞免疫表型分析、促炎细胞因子测定和外周血凋亡测量的免疫学检测可能是一种有用的工具，有助于更深入了解AD的发病机制以及免疫激活水平，而免疫激活水平可能是个体AD患者淋巴细胞病理状态的特征。
