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膜蛋白在生长受抑制的Daudi细胞中受α干扰素刺激,但在抗性Daudi细胞中不受刺激。

Membrane proteins stimulated by interferon-alpha in growth-inhibited, but not in resistant, Daudi cells.

作者信息

Quek A, Goding J, Hoogenraad N

机构信息

Department of Biochemistry, La Trobe University, Bundoora, Vic., Australia.

出版信息

Immunol Cell Biol. 1988 Oct-Dec;66 ( Pt 5-6):353-9. doi: 10.1038/icb.1988.46.

Abstract

Interferon-alpha (alpha-IFN) stimulates the expression of a number of plasma membrane proteins in wild-type Daudi cells. These proteins are not induced in a variant subline of Daudi cells, which is insensitive to growth inhibition, nor in another which is partially growth-inhibited by alpha-IFN. Two-dimensional polyacrylamide gel electrophoresis of [125I]-surface labelled Daudi membrane proteins indicates that the major protein induced de novo has a molecular weight of 16 kD (p16), and proteins of 20 kD (p20), 28 kD (p28) and 106 kD (p106) which are present in untreated cells are also induced by alpha-IFN. A monoclonal antibody was produced against p106 by immunizing mice with cells treated for 16 h with alpha-IFN followed by differential screening against treated and untreated cells. The antibody (A96/G8) immunoprecipitated p106 from cell lysates and was used to show that p106 is induced approximately two-fold by alpha-IFN in wild-type Daudi cells, but not at all in Daudi cells which show a complete or partial resistance to growth inhibition by alpha-IFN; gamma-IFN does not induce p16 or p106 and also does not inhibit cell growth. The induction of p16 and p106 precedes the observed inhibition of growth by IFN. The levels of p16 and p106 are elevated by 8.5 h and in the case of p16 continue to rise for 16 h after the addition of IFN, whereas p106 reaches a peak by 8.5 h. In contrast, growth inhibition is not observed by 8 h and does not become clear-cut until 24 h after the addition of IFN.

摘要

α-干扰素(α-IFN)可刺激野生型Daudi细胞中多种质膜蛋白的表达。这些蛋白在对生长抑制不敏感的Daudi细胞变异亚系中未被诱导,在另一个对α-IFN有部分生长抑制作用的亚系中也未被诱导。对经[125I]表面标记的Daudi膜蛋白进行二维聚丙烯酰胺凝胶电泳表明,新诱导产生的主要蛋白分子量为16kD(p16),未处理细胞中存在的20kD(p20)、28kD(p28)和106kD(p106)蛋白也被α-IFN诱导。通过用α-IFN处理16小时后的细胞免疫小鼠,然后对处理过和未处理的细胞进行差异筛选,制备了一种针对p106的单克隆抗体。该抗体(A96/G8)可从细胞裂解物中免疫沉淀p106,用于表明在野生型Daudi细胞中,α-IFN可使p106诱导增加约两倍,但在对α-IFN生长抑制完全或部分耐药的Daudi细胞中则完全不诱导;γ-IFN不诱导p16或p106,也不抑制细胞生长。p16和p106的诱导先于IFN观察到的生长抑制。加入IFN后8.5小时,p16和p106的水平升高,对于p16而言,在加入IFN后16小时仍继续上升,而p106在8.5小时达到峰值。相比之下,在8小时时未观察到生长抑制,直到加入IFN后24小时才变得明显。

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