Characterization of epidermal cytokine profiles in sensitization and elicitation phases of allergic contact dermatitis as well as irritant contact dermatitis in mouse skin.

Lymphokine and cytokine research Pub Date : 1994-12-01
S Kondo, S Pastore, G M Shivji, R C McKenzie, D N Sauder
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Abstract

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)

小鼠皮肤过敏性接触性皮炎和刺激性接触性皮炎致敏和诱发期表皮细胞因子谱的表征。
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)和刺激性接触性皮炎(ICD)致敏和激发阶段的初始分子事件。用0.5%二硝基氟苯(DNFB)致敏BALB/c小鼠背部皮肤,6天后用0.2% DNFB刺激耳部,引起过敏性接触性超敏反应(ACDe)。为了从同一解剖区域检查致敏阶段的细胞因子谱,其他动物在耳朵而不是背部皮肤上致敏。用0.5% DNFB涂敷幼鼠耳,诱导ACD致敏期。将十二烷基硫酸钠(SLS)作为刺激对照,应用于另一组小鼠耳部诱导ICD。在每次处理后的不同时间点从处理过的耳朵表皮提取总RNA,逆转录为cDNA,并使用放射性标记的细胞因子特异性引物进行PCR扩增。扩增产物通过电泳和放射自显影测定大小,并通过密度测定半定量。射线自显像相对于-肌动蛋白信号归一化。ACDs和ACDe细胞因子mRNA表达模式相似;也就是说,在半抗原应用后6小时,白细胞介素(IL)-1 β、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,这种上调一直持续到治疗后24小时。(摘要删节250字)
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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