Lohoff M, Prechtl S, Sommer F, Roellinghoff M, Schmitt E, Gradehandt G, Rohwer P, Stride B D, Cole S P, Deeley R G
Institut für Klinische Mikrobiologie und Immunologie, University of Erlangen, 91054 Erlangen, Germany.
J Clin Invest. 1998 Feb 1;101(3):703-10. doi: 10.1172/JCI824.
A transmembrane pump for organic anions was identified in resting murine T helper (Th) 2, but not Th1 lymphocyte cell clones, as revealed by extrusion of a fluorescent dye. Dye extrusion inhibition studies suggested that the pump may be the multidrug-resistance protein (MRP). The different expression of the pump in resting Th1 and Th2 cell clones correlated with their respective levels of MRP mRNA. The pump was inducible in Th1 cells by antigenic stimulation in vitro leading to equal expression in activated Th1 and Th2 cell clones. This suggested that dye extrusion might allow the detection of Th2 (resting or activated) or of activated Th1 cells ex vivo based on a functional parameter. To test this, mice were infected with Leishmania major parasites to activate L. major-specific T cells of either Th1 (C57BL/6 mice) or Th2 (BALB/c mice) phenotype: 2-3% of CD4+ lymph node T cells of both strains of mice extruded the dye, defining a cell subset that did not coincide with subsets defined by other activation markers. Fluorescence-activated cell-sorting revealed that the lymphokine response (Th1 or Th2, respectively) to L. major antigens was restricted to this dye-extruding subset.
通过荧光染料的外排发现,在静息的小鼠辅助性T细胞(Th)2而非Th1淋巴细胞克隆中存在一种有机阴离子跨膜泵。染料外排抑制研究表明,该泵可能是多药耐药蛋白(MRP)。静息的Th1和Th2细胞克隆中该泵的不同表达与其各自的MRP mRNA水平相关。体外抗原刺激可诱导Th1细胞中的该泵表达,导致活化的Th1和Th2细胞克隆中表达水平相等。这表明基于功能参数,染料外排可能允许在体外检测Th2(静息或活化)或活化的Th1细胞。为了验证这一点,用硕大利什曼原虫感染小鼠以激活Th1(C57BL/6小鼠)或Th2(BALB/c小鼠)表型的硕大利什曼原虫特异性T细胞:两种品系小鼠的2-3%的CD4+淋巴结T细胞可外排染料,确定了一个与由其他活化标志物定义的亚群不一致的细胞亚群。荧光激活细胞分选显示,对硕大利什曼原虫抗原的淋巴因子反应(分别为Th1或Th2)仅限于这个染料外排亚群。