摘要
目的检测人角蛋白17对银屑病患者T细胞增殖和细胞因子分泌的影响。方法从银屑病皮损中提取总RNA,反转录成cDNA,通过PCR方法扩增人角蛋白17基因,利用GST融合表达载体pGEX-4T-1表达相应融合蛋白;梯度离心和绵羊红细胞花环沉淀法分离纯化T细胞;直接活细胞计数和MTT法检测T细胞增殖程度,ELISA检测培养上清液中干扰素γ浓度。结果和正常人对照相比,角蛋白17可以显著促进银屑病患者T细胞的增殖,并且促进其分泌干扰素γ(P值均<0.001)。结论角蛋白17对银屑病患者T细胞功能的促进作用提示其可能是引发银屑病的重要自身抗原之一。
Objective To investigate the immunological effects of human keratin 17 (K17) on T lymphocytes from psoriatic patients. Methods Total RNA was extracted from the epidermis of psoriatic lesions, and mRNA was reversely transcribed to cDNA. The K17 gene was amplified by PCR and inserted into pGEX-4T-1 expression vector, and then K17 was expressed and purified. T cells were isolated and purified by gra-dient centrifugation and AET-SRBC (2-aminoethylosothiouronium bromide-treated sheep red blood cells) precipitation. MTT assay and direct cell counting method were used to detect the proliferation of T cells. The concentration of IFN- in culture supernatant was measured by ELISA. Results Compared with the control group, K17 significantly enhanced the proliferation of T cells from psoriatic patients and increased the expression level of IFN- (P < 0.001 respectively). Conclusion K17 can enhance the function of T cells in psoriatic patients, and it may play a role as an autoantigen in the immunological pathogenesis of psoriasis.
出处
《中华皮肤科杂志》
CAS
CSCD
北大核心
2004年第11期659-661,共3页
Chinese Journal of Dermatology
基金
国家自然科学基金(30371650)
