Cloning of mouse Ox40: a T cell activation marker that may mediate T-B cell interactions.
Cloning of mouse Ox40: a T cell activation marker that may mediate T-B cell interactions.
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DOI：
10.4049/jimmunol.151.10.5261
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发表时间：
1993-11
影响因子：
4.4
通讯作者：
D. Calderhead;J. Buhlmann;A. V. D. van den Eertwegh;E. Claassen;R. Noelle;H. P. Fell
中科院分区：
文献类型：
--
作者：
D. Calderhead;J. Buhlmann;A. V. D. van den Eertwegh;E. Claassen;R. Noelle;H. P. Fell
从小鼠Th细胞系Th 2 D.10制备cDNA文库，并通过聚合酶链反应用于克隆小鼠Ox 40同源物。小鼠序列与大鼠序列的比较显示，在DNA和蛋白质水平上，两个序列之间的同源性大于90%。北方印迹分析发现，在大鼠中，Ox 40的表达似乎仅限于活化的T细胞。制备嵌合受体球蛋白以包括与人IgG 1的铰链-CH 2-CH 3结构域偶联的小鼠Ox 40胞外结构域（Ox 40-IG）。这种分子的可溶性形式然后用于鉴定携带Ox 40配体的细胞。流式细胞仪分析显示，Ox 40-IG结合到腹膜B细胞的一个子集，以及一部分的LPS激活的脾B细胞。使用Ag特异性缀合物和Ox 40-IG对脾切片进行免疫染色，发现显著比例的抗体形成细胞与Ox 40-IG共染色。细胞表面放射性标记的腹膜B细胞的免疫沉淀表明与70 kDa蛋白质的特异性相互作用。
A cDNA library was prepared from the murine Th cell line Th2 D.10 and used to clone the murine homologue of Ox40 by polymerase chain reaction. Comparison of the mouse sequence with the rat revealed greater than 90% homology between the two sequences at both the DNA and protein level. Northern blot analysis found that, as in the rat, Ox40 expression appears to be restricted to activated T cells. A chimeric receptor globulin was prepared to include the mouse Ox40 extracellular domain coupled to the hinge-CH2-CH3 domains of human IgG1 (Ox40-Ig). This soluble form of the molecular was then used to identify cells bearing a ligand for Ox40. FACS analysis revealed that Ox40-Ig bound to a subset of peritoneal B cells as well as to a fraction of LPS-activated splenic B cells. Immunostaining of spleen sections using an Ag-specific conjugate and Ox40-Ig found a significant proportion of antibody-forming cells co-stained with Ox40-Ig. Immunoprecipitation of cell-surface radiolabeled peritoneal B cells suggests a specific interaction with a protein of 70 kDa.