Expression and co-cytokine function of murine thioredoxin adult T cell leukaemia-derived factor (ADF)
Expression and co-cytokine function of murine thioredoxin adult T cell leukaemia-derived factor (ADF)
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DOI：
10.1006/cyto.1996.0002
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发表时间：
1996-01-01
期刊：
影响因子：
3.8
通讯作者：
Gessner, A
中科院分区：
文献类型：
--
作者：
Blum, H;Rollinghoff, M;Gessner, A
人ADF(成人T细胞白血病衍生因子)是硫氧还蛋白的一种异构体，可促进某些人淋巴样细胞系的增殖，并参与许多硫醇依赖的还原反应。为了研究小鼠ADF同源物的功能，我们建立了小鼠ADF在大肠杆菌中的诱导性过表达，并建立了一种纯化方法，从而获得了明显均一的14 kDa蛋白。用小鼠Th2细胞(D10、G4、1)和CTLL-2细胞对重组ADF进行增殖实验。在IL-2、IL-4、IL-7和IL-9协同作用下，ADF具有协同细胞因子活性。亲和纯化的兔抗ADF多克隆抗血清可中和上述促增殖作用。ADF的作用严重依赖于2-巯基乙醇的存在。细菌硫氧还蛋白对小鼠T细胞的增殖也有类似的作用。因此，这些蛋白质与硫醇相关的还原能力对于它们的生长促进活性是必不可少的。在几个小鼠细胞克隆和细胞系以及小鼠组织中，ADF在mRNA和蛋白质水平上普遍表达。竞争性聚合酶链式反应证实，与细胞因子mRNAs(如IL-4和IL-13)相比，小鼠Th2克隆体和脾细胞的ADF mRNA的表达不受T细胞受体复合体刺激的影响。因此，小鼠ADF是一种在多种细胞中结构性表达的蛋白质，具有增强几种细胞因子对小鼠T细胞的增殖作用的能力。(C)1996年学术出版社有限公司
Human ADF (adult T cell leukaemia-derived factor), an isoform of thioredoxin, promotes proliferation of certain human lymphoid cell lines and is involved in many thiol-dependent reducing reactions. To study functional aspects of the murine homologue, we established inducible overexpression of murine ADF in E, coli and a purification method which led to an apparently homogeneous 14 kDa protein. This recombinant ADF was tested in proliferation assays with murine Th2 cells (D10,G4,1) and CTLL-2 cells. In synergy with IL-2, IL-4, IL-7 and IL-9 ADF displayed co-cytokine activity. These proliferative effects were neutralized by an affinity-purified polyclonal rabbit anti-ADF antiserum. The effects of ADF were critically dependent on the presence of 2-mercaptoethanol. Bacterial thioredoxin had similar effects on the proliferation of murine T cells. Thus, the thiol-related reducing capacity of these proteins is essential for their growth promoting activity. As investigated at the levels of mRNA and protein in several murine cell clones and lines as well as in mouse tissues ADF is expressed ubiquitously. Finally it could be demonstrated by competitive PCR that in contrast to cytokine mRNAs (e,g, IL-4 and IL-13) the expression of ADF mRNA in murine Th2 clones and spleen cells is not influenced by stimulation of these cells through the T cell receptor complex. Murine ADF therefore represents a protein constitutively expressed in a wide variety of cells,vith the capacity to enhance the proliferative effect of several cytokines on murine T cells. (C) 1996 Academic Press Limited