Lavergne S N, Wang H, Callan H E, Park B K, Naisbitt D J
Department of Pharmacology, Centre for Drug Safety Science, The University of Liverpool, Liverpool, UK.
J Pharmacol Exp Ther. 2009 Nov;331(2):372-81. doi: 10.1124/jpet.109.155374. Epub 2009 Aug 7.
Antigen-presenting cells (APC) are thought to play an important role in the pathogenesis of drug-induced immune reactions. Various pathological factors can activate APC and therefore influence the immune equilibrium. It is interesting that several diseases have been associated with an increased rate of drug allergy. The aim of this project was to evaluate the impact of such "danger signals" on sulfamethoxazole (SMX) metabolism in human APC (peripheral blood mononuclear cells, Epstein-Barr virus-modified B lymphocytes, monocyte-derived dendritic cells, and two cell lines). APC were incubated with SMX (100 microM-2 mM; 5 min-24 h), in the presence of pathological factors: bacterial endotoxins (lipopolysaccharide and staphylococcal enterotoxin B), flu viral proteins, cytokines [interleukin (IL)-1beta, IL-6, IL-10; tumor necrosis factor-alpha; interferon-gamma; and transforming growth factor-beta], inflammatory molecules (prostaglandin E2, human serum complement, and activated protein C), oxidants (buthionine sulfoximine and H(2)O(2)), and hyperthermia (37.5-39.5 degrees C). Adduct formation was evaluated by enzyme-linked immunosorbent assay and confocal microscopy. SMX-protein adduct formation was time- and concentration-dependent for each cell type tested, in both physiological and danger conditions. A danger environment significantly increased the formation of SMX-protein adducts and significantly shortened the delay for their detection. An additive effect was observed with a combination of danger signals. Dimedone (chemical selectively binding cysteine sulfenic acid) and antioxidants decreased both baseline and danger-enhanced SMX-adduct formation. Various enzyme inhibitors were associated with a significant decrease in SMX-adduct levels, with a pattern varying depending on the cell type and the culture conditions. These results illustrate that danger signals enhance the formation of intracellular SMX-protein adducts in human APC. These findings might be relevant to the increased frequency of drug allergy in certain disease states.
抗原呈递细胞（APC）被认为在药物诱导的免疫反应发病机制中起重要作用。多种病理因素可激活APC，进而影响免疫平衡。有趣的是，几种疾病与药物过敏率增加有关。本项目的目的是评估此类“危险信号”对人APC（外周血单核细胞、爱泼斯坦-巴尔病毒修饰的B淋巴细胞、单核细胞衍生的树突状细胞和两种细胞系）中磺胺甲恶唑（SMX）代谢的影响。将APC与SMX（100微摩尔/升 - 2毫摩尔/升；5分钟 - 24小时）一起孵育，同时存在以下病理因素：细菌内毒素（脂多糖和葡萄球菌肠毒素B）、流感病毒蛋白、细胞因子[白细胞介素（IL）-1β、IL-6、IL-10；肿瘤坏死因子-α；干扰素-γ；以及转化生长因子-β]、炎症分子（前列腺素E2、人血清补体和活化蛋白C）、氧化剂（丁硫氨酸亚砜胺和H₂O₂）和热疗（37.5 - 39.5摄氏度）。通过酶联免疫吸附测定和共聚焦显微镜评估加合物的形成。在生理和危险条件下，对于每种测试的细胞类型，SMX-蛋白质加合物的形成均呈时间和浓度依赖性。危险环境显著增加了SMX-蛋白质加合物的形成，并显著缩短了检测到它们的延迟时间。观察到危险信号组合具有累加效应。二甲酮（化学选择性结合半胱氨酸亚磺酸）和抗氧化剂可降低基线和危险增强的SMX-加合物形成。各种酶抑制剂与SMX-加合物水平的显著降低有关，其模式因细胞类型和培养条件而异。这些结果表明，危险信号增强了人APC中细胞内SMX-蛋白质加合物的形成。这些发现可能与某些疾病状态下药物过敏频率增加有关。
