Kondo S, Pastore S, Shivji G M, McKenzie R C, Sauder D N
Division of Dermatology, Sunnybrook Health Science Centre, University of Toronto, Ontario, Canada.
Lymphokine Cytokine Res. 1994 Dec;13(6):367-75.
Epidermal cytokines are known to participate in the initiation of immune and inflammatory processes in the skin. In the present study, we examined epidermal cytokine mRNA levels to elucidate the initial molecular events in the sensitization and elicitation phases of allergic contact dermatitis (ACD) as well as in irritant contact dermatitis (ICD). BALB/c mice were sensitized on the dorsal skin with 0.5% dinitrofluorobenzene (DNFB) and challenged with 0.2% DNFB on the ears 6 days later to elicit allergic contact hypersensitivity (ACDe), the elicitation phase. To examine cytokine profiles during the sensitization phase from the same anatomic area, other animals were sensitized on ear instead of dorsal skin. The sensitization phase of ACD (ACDs) was induced by painting the ears of naive mice with 0.5% DNFB. Sodium lauryl sulfate (SLS), utilized as an irritant control, was also applied to the ears of another group of mice to induce ICD. Total RNA was extracted from the epidermis of the treated ears at various time points after each treatment, reverse transcribed to cDNA, and amplified by PCR using radiolabeled cytokine-specific primers. Amplified products were sized by electrophoresis and autoradiography and semiquantitated by densitometry. Autoradiographs were normalized relative to beta-actin signals. ACDs and ACDe showed similar patterns of cytokine mRNA levels; that is, at 6 h after hapten application, interleukin (IL)-1 beta, IL-6, IL-10, and granulocyte-macrophage colony-stimulating factor (GM-CSF) mRNA levels were upregulated, and this upregulation was observed until 24 h after treatment.(ABSTRACT TRUNCATED AT 250 WORDS)
已知表皮细胞因子参与皮肤免疫和炎症过程的启动。在本研究中，我们检测了表皮细胞因子mRNA水平，以阐明过敏性接触性皮炎（ACD）以及刺激性接触性皮炎（ICD）致敏和激发阶段的初始分子事件。用0.5%二硝基氟苯（DNFB）对BALB/c小鼠背部皮肤进行致敏，6天后用0.2%DNFB刺激耳部以引发过敏性接触超敏反应（ACDe），即激发阶段。为了检测来自同一解剖区域致敏阶段的细胞因子谱，对其他动物的耳部而非背部皮肤进行致敏。用0.5%DNFB涂抹未致敏小鼠的耳部诱导ACD的致敏阶段（ACDs）。将十二烷基硫酸钠（SLS）用作刺激对照，也应用于另一组小鼠的耳部以诱导ICD。在每次处理后的不同时间点，从处理过的耳部表皮中提取总RNA，逆转录为cDNA，并使用放射性标记的细胞因子特异性引物通过PCR进行扩增。扩增产物通过电泳和放射自显影进行大小测定，并通过光密度测定进行半定量。放射自显影片相对于β-肌动蛋白信号进行标准化。ACDs和ACDe显示出相似的细胞因子mRNA水平模式；也就是说，在应用半抗原后6小时，白细胞介素（IL）-1β、IL-6、IL-10和粒细胞-巨噬细胞集落刺激因子（GM-CSF）mRNA水平上调，并且这种上调在处理后24小时内都能观察到。（摘要截断于250字）
