Reijonen Helena, Novak Erik J, Kochik Sharon, Heninger Anne, Liu Andrew W, Kwok William W, Nepom Gerald T
Virginia Mason Research Center, Benaroya Research Institute, Seattle, Washington 98101-2795, USA.
Diabetes. 2002 May;51(5):1375-82. doi: 10.2337/diabetes.51.5.1375.
Soluble HLA-DR401 or -DR404 tetramers containing a peptide corresponding to an immunodominant epitope from human GAD65 were used to analyze peripheral blood T-cells of newly diagnosed type 1 diabetic patients and at-risk subjects. Peripheral blood mononuclear cells were expanded on antigen-presenting cells presenting GAD65 peptide and subsequently activated with specific plate-bound class II-peptide monomers. T-cell activation defined in flow cytometry by CD4(high) and/or CD25 markers were observed in all type 1 diabetic patients and some at-risk subjects, but not in normal control subjects. The activated T-cells stained positive with tetramers containing the GAD65 epitope 555-567. Tetramer-positive cells were CD4(high) T-cells with high avidity for an immunodominant GAD65 T-cell epitope. Phenotyping of T-cells utilizing HLA class II tetramers provides a new tool to characterize the autoimmune response in type 1 diabetes.
含有对应于人谷氨酸脱羧酶65（GAD65）免疫显性表位肽段的可溶性HLA - DR401或 - DR404四聚体，被用于分析新诊断的1型糖尿病患者和高危个体的外周血T细胞。外周血单个核细胞在呈递GAD65肽段的抗原呈递细胞上进行扩增，随后用特异性板结合II类肽单体激活。在所有1型糖尿病患者和一些高危个体中观察到通过流式细胞术由CD4（高表达）和/或CD25标志物定义的T细胞活化，但在正常对照个体中未观察到。活化的T细胞用含有GAD65表位555 - 567的四聚体染色呈阳性。四聚体阳性细胞是对免疫显性GAD65 T细胞表位具有高亲和力的CD4（高表达）T细胞。利用HLA II类四聚体对T细胞进行表型分析为表征1型糖尿病中的自身免疫反应提供了一种新工具。
