Cancer Research Center, (IBMCC; CSIC-USAL), IBSAL and Cytometry Service, University of Salamanca, Salamanca, Spain.
Alcohol Clin Exp Res. 2013 Aug;37(8):1361-9. doi: 10.1111/acer.12095. Epub 2013 Mar 29.
Development of alcoholic hepatitis (AH) may be favored by the activation of the innate immune response. Recently, decreased numbers of circulating regulatory T cells (Tregs) have been reported in diseases associated with an immune activation status, but no studies have focused so far, in investigating the distribution of Tregs in chronic alcoholism and its potential association with liver disease. Here, we analyzed for the first time the frequency of peripheral blood (PB) Tregs and Treg subsets in AH and its relationship with the production of inflammatory cytokines by PB monocytes and dendritic cells (DCs).
PB samples from 25 male patients with AH were studied; in parallel, 15 male chronic alcoholic patients without liver disease (AWLD) and 17 male healthy donors were also studied, as controls. The distribution of CD4⁺CD25hiCD127-/lo Tregs and their maturation subsets (naïve, central memory, and peripheral memory Tregs) was analyzed by flow cytometry. Spontaneous and in vitro-stimulated production of inflammatory cytokines by PB monocytes and DCs was analyzed by flow cytometry at the cytoplasmic level.
Patients with AH showed decreased (p < 0.05) numbers of PB CD4⁺CD25hiCD127-/lo Tregs at the expense of all maturation-associated subsets, while AWLD and healthy subjects showed a similar (p > 0.05) distribution of PB CD4⁺CD25hiCD127-/lo Tregs. Interestingly, significantly increased amounts of spontaneously produced inflammatory cytokines were found among circulating monocyte-derived DCs and monocytes from AH (and AWLD) patients in comparison with healthy donors. Conversely, the ability of these cell subsets to produce cytokines after in vitro stimulation was lower (p < 0.05) in AH versus the 2 control groups.
PB CD4⁺CD25hiCD127-/lo Tregs are significantly decreased in patients with AH when compared to both healthy and AWLD; this may contribute to explain the more pronounced activation of the innate immune response observed in AH, as reflected by an increased secretion of inflammatory cytokines by PB DCs and monocytes, and could facilitate the development of liver disease.
酒精性肝炎（AH）的发展可能有利于先天免疫反应的激活。最近，在与免疫激活状态相关的疾病中，循环调节性 T 细胞（Tregs）的数量减少，但迄今为止尚无研究集中在慢性酒精中毒中 Tregs 的分布及其与肝病的潜在关联。在这里，我们首次分析了外周血（PB）Tregs 和 Treg 亚群在 AH 中的频率，并研究了其与 PB 单核细胞和树突状细胞（DC）产生炎症细胞因子的关系。
研究了 25 名男性 AH 患者的 PB 样本；同时，还研究了 15 名男性慢性酒精性肝病（AWLD）患者和 17 名男性健康供体作为对照。通过流式细胞术分析 CD4⁺CD25hiCD127-/lo Tregs 及其成熟亚群（幼稚、中央记忆和外周记忆 Tregs）的分布。通过流式细胞术在细胞质水平分析 PB 单核细胞和 DC 自发和体外刺激产生炎症细胞因子的情况。
AH 患者 PB CD4⁺CD25hiCD127-/lo Tregs 数量减少（p < 0.05），所有成熟相关亚群均受累，而 AWLD 和健康受试者 PB CD4⁺CD25hiCD127-/lo Tregs 分布相似（p > 0.05）。有趣的是，与健康供体相比，AH（和 AWLD）患者循环单核细胞衍生 DC 和单核细胞中自发产生的炎症细胞因子量显著增加。相反，与这两个对照组相比，这些细胞亚群在体外刺激后产生细胞因子的能力较低（p < 0.05）。
与健康和 AWLD 相比，AH 患者的 PB CD4⁺CD25hiCD127-/lo Tregs 明显减少；这可能有助于解释 AH 中观察到的先天免疫反应更明显的激活，这反映在 PB DC 和单核细胞中炎症细胞因子的分泌增加，并且可能促进肝病的发展。
