Laso F J, Madruga J I, San Miguel J F, Ciudad J, López A, Alvarez Mon M, Orfao A
Servicio de Medicina Interna II, Hospital Universitario, Universidad de Salamanca, Spain.
Cytometry. 1996 Dec 15;26(4):275-80. doi: 10.1002/(SICI)1097-0320(19961215)26:4<275::AID-CYTO6>3.0.CO;2-B.
The aim of the present study was to analyze on chronic alcoholic patients the effect of ethanol (EtOH) withdrawal on the immune system through the investigation of the distribution of PB lymphoid subsets, using multiple-stainings with monoclonal antibodies and flow cytometry. For this purpose a group of 20 patients with active alcoholism without liver disease, negative for hepatitis virus, and without malnutrition was analyzed and followed for 9 months after alcohol consumption had been discontinued. Twenty-five age- and sex-matched healthy volunteers were included in the study. The following panel of monoclonal antibodies combinations (FITC/PE/PerCP or PE-Cy5) was used: TCR alpha beta/CD3/HLA DR, CD25/CD56/CD3, TCR gamma delta/CD3/HLA DR, CD45RA/CD45R0/CD4, CD3/CD8, CD19/CD5, and CD3/CD11c. Analysis was performed on at least 1,500 events/tube at flow cytometry using the Lysys II software program. During the alcohol intake period, the most striking findings were a significant (P < 0.05) expansion of the CD8+ T-lymphocyte subset, which coexpresses the activation associated antigens HLA DR and CD11c, as well as a significant increase in both NK-cells (CD3-/CD56+) and the T-cell subset with NK activity coexpressing CD3 and CD56 (P < 0.05 and P < 0.01, respectively). In addition, a decrease in the CD5+ B-cells (P < 0.05), associated with reduced serum gamma-globulin levels, was also observed. During alcohol withdrawal, a rapid decrease towards normal values of activated CD8+/HLA DR+ and CD11c+ T-lymphocytes was observed as well as a normalization of CD19+/CD5+ B-cells and gamma-globulin serum levels; these changes might be directly related to EtOH suppression. Surprisingly, however, new immunological imbalances emerged in spite of the absence of alcohol intake. Thus, a progressive and significant expansion (P < 0.05) of CD4+ T-cells associated with an increased expression of the CD25 activation-related antigen and a preferential use of the CD45R0 isoform by CD4+ T-cells were observed. In parallel, there was an even more evident increase (P < 0.01) in the number of PB NK-cells. Our results show that EtOH consumption induces changes in the immune system, its effects persisting or even becoming more evident after suppression of EtOH intake for a 9 month period.
本研究的目的是通过使用单克隆抗体多染色和流式细胞术研究外周血淋巴细胞亚群的分布,分析慢性酒精中毒患者乙醇(EtOH)戒断对免疫系统的影响。为此,对一组20例无肝病、肝炎病毒阴性且无营养不良的酒精依赖活跃期患者进行了分析,并在戒酒9个月后进行随访。25名年龄和性别匹配的健康志愿者纳入研究。使用了以下单克隆抗体组合面板(FITC/PE/PerCP或PE-Cy5):TCRαβ/CD3/HLA DR、CD25/CD56/CD3、TCRγδ/CD3/HLA DR、CD45RA/CD45R0/CD4、CD3/CD8、CD19/CD5和CD3/CD11c。使用Lysys II软件程序在流式细胞仪上对每个样本至少分析一千五百个事件。在饮酒期间,最显著的发现是共表达激活相关抗原HLA DR和CD11c的CD8 + T淋巴细胞亚群显著(P < 0.05)扩增,以及自然杀伤细胞(CD3-/CD56 +)和共表达CD3和CD56的具有自然杀伤活性的T细胞亚群均显著增加(分别为P < 0.05和P < 0.01)。此外,还观察到CD5 + B细胞减少(P < 0.05),同时血清γ球蛋白水平降低。在戒酒期间,观察到激活的CD8 + /HLA DR +和CD11c + T淋巴细胞迅速降至正常值,以及CD19 + /CD5 + B细胞和γ球蛋白血清水平恢复正常;这些变化可能与乙醇抑制直接相关。然而,令人惊讶的是,尽管不再饮酒,新的免疫失衡仍出现。因此,观察到CD4 + T细胞逐渐且显著(P < 0.05)扩增,同时与CD25激活相关抗原表达增加以及CD4 + T细胞优先使用CD45R0亚型有关。与此同时,外周血自然杀伤细胞数量有更明显的增加(P < 0.01)。我们的结果表明,饮酒会引起免疫系统变化,在戒酒9个月后,其影响仍然存在甚至更加明显。