Slobodin Gleb, Kessel Aharon, Peri Regina, Zaigraikin Natalia, Rozenbaum Michael, Rosner Itzhak, Toubi Elias
Department of Internal Medicine A, Bnai Zion Medical Center, Ruth and Bruce Rappaport Faculty of Medicine, Technion, Haifa, Israel.
Inflammation. 2009 Jun;32(3):146-50. doi: 10.1007/s10753-009-9113-7.
The success of anti-tumor necrosis factor alpha (TNFalpha) therapies has led to increased interest as to the mechanisms and consequences of TNFalpha blockade. The aim of the study was to examine the effects of TNFalpha blockade by etanercept on lipopolysaccharide (LPS) or peptidoglycan (PG)-induced maturation of human monocyte-derived dendritic cells (MDDCs). MDDCs grown from peripheral blood of healthy donors were stimulated by LPS or PG with/without the presence of etanercept. Concentrations of TNFalpha in cell supernatants were assessed by ELISA, while the cells were stained with monoclonal antibodies to CD83, CD80, CD86, CD11c, CD40, HLA-DR, and annexin-V and acquired using a flow cytometer. Etanercept significantly decreased the stimulated cell surface expression of HLA-DR, CD80, CD86, CD40 and CD83 on MDDCs in all examined samples. Etanercept in the same dose, but denatured to loss of specificity for TNFalpha, failed to change any of the aforementioned markers. In the presence of etanercept, concentrations of TNFalpha in cell supernatants were decreased by 53% on average, with a range of 25%-87%. Etanercept impaired the stimulated maturation of MDDCs by neutralizing the induced TNFalpha, produced by the same MDDCs after antigenic stimulation. The reported data confirms that TNFalpha blockade may have a direct effect on DCs, with a wide spectrum of potential secondary effects downstream. The data also suggests the presence of TNFalpha-mediated autocrine signaling, serving to accelerate or catalyze the maturation process of MDDCs.
抗肿瘤坏死因子α（TNFα）疗法的成功引发了人们对TNFα阻断机制及后果的更多关注。本研究的目的是检测依那西普阻断TNFα对脂多糖（LPS）或肽聚糖（PG）诱导的人单核细胞衍生树突状细胞（MDDC）成熟的影响。从健康供体外周血培养的MDDC在有或无依那西普存在的情况下，用LPS或PG进行刺激。通过酶联免疫吸附测定（ELISA）评估细胞上清液中TNFα的浓度，同时用针对CD83、CD80、CD86、CD11c、CD40、HLA-DR和膜联蛋白-V的单克隆抗体对细胞进行染色，并使用流式细胞仪采集数据。在所有检测样本中，依那西普显著降低了MDDC上HLA-DR、CD80、CD86、CD40和CD83的刺激细胞表面表达。相同剂量但对TNFα失去特异性的变性依那西普未能改变上述任何标志物。在依那西普存在的情况下，细胞上清液中TNFα的浓度平均降低了53%，范围为25% - 87%。依那西普通过中和抗原刺激后同一MDDC产生的诱导性TNFα，损害了MDDC的刺激成熟。报告的数据证实，TNFα阻断可能对树突状细胞有直接影响，并在下游产生广泛的潜在继发效应。数据还表明存在TNFα介导的自分泌信号，有助于加速或催化MDDC的成熟过程。
