아토피피부염 환자에서 인터루킨-12에 의한 CD4+ 세포 단백 변화에 관한 프로테옴 분석

Abstract

The pathogenesis of atopic dermatitis is based on an inflammatory mechanism involving type 2 cytokines such as interleukin (IL)-4 and IL-13. CD4+ T cells in atopic dermatitis express predominantly T helper (Th)2 phenotype and they down-regulates IL-12Rβ2. IL-12 is a major cytokine in the differentiation of naive CD4+ T cells into Th1 cells. This mechanism is closely related with the expression of IL-12Rβ2. However, it remains unclear that IL-12 signaling mechanism in polarized Th2 cells. The aim of this study was to identify IL-12 responsiveness in CD4+ T cells of patients with atopic dermatitis using a proteomic tools. CD4+ T cells isolated from peripheral blood of patients with atopic dermatitis were treated with neutralizing anti-IL-4 antibody (200 ng/ml) and IL-12 (2 ng/ml) and parallel cultures of untreated cells were also prepared. On day 3, surface phenotype change was examined using IL-12Rβ2 antibody by FACS analysis and separate CD4+ T cells by two-dimensional electrophoresis (2-DE). About 1500 protein spots were detected in the 2-DE gels by modified silver staining. Several areas of the 2-DE map exhibited quantitative and qualitative changes after IL-12 treatment. Several regions including actins showed variations according to the samples. A group of three spots detected in the area of pI 6.0-10.0 with molecular weight about 40 kDa. In addition, decreased or increased spots were observed in the regions of pI 6.5-7.0 with molecular weight about 20 kDa. The identification of these spots must be established in further study in order to find targets regulated by IL-12 in CD4+ T cells of atopic patients.