Functional and inducible expression of a transfected murine class II major histocompatibility complex gene.

Functional and inducible expression of a transfected murine class II major histocompatibility complex gene.
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转染的小鼠 II 类主要组织相容性复合体基因的功能和诱导表达。

DOI:
10.1073/pnas.81.7.2045
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发表时间:
1984
影响因子:
11.1
通讯作者:
Tonegawa,S
Tonegawa,S
中科院分区:
综合性期刊1区
文献类型:
--
作者:
Folsom,V;Gold,DP;White,J;Marrack,P;Kappler,J;Tonegawa,S

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利用原生质球融合技术,我们将克隆的E β B基因导入两个d单倍型细胞系,B淋巴瘤细胞系A20- 2 J和巨噬细胞肿瘤细胞系P388 D1。单克隆抗体分析表明,转染的E β B基因产物与内源性E α链结合形成E α dE β B复合物。虽然E α dE β B b的表达在用E β B基因转染的A20- 2 J细胞中是组成性的,但仅在用来自伴刀豆球蛋白A(Con A)刺激的T细胞的培养上清液处理细胞后,才在转染的巨噬细胞中检测到E α dE β B b的表面表达。转染的B淋巴瘤细胞和转染的Con A上清液处理的巨噬细胞获得了将抗原呈递给E α dE β b限制性T细胞杂交瘤的能力。在巨噬细胞宿主中观察到的转染基因的诱导型表达表明,负责调节E β B基因表达的序列可能与转染基因相关。与定向诱变相结合,本文所述的系统提供了一种研究(i)E β B序列的方法,该序列在确定E分子的限制特异性中很重要,和(ii)与E β基因相关的序列,该序列可能在调节E β链表达中很重要。
Using the spheroplast fusion technique, we have introduced the cloned E beta b gene into two d haplotype cell lines, the B lymphoma line A20-2J and the macrophage tumor line P388D1. Analysis with a monoclonal antibody indicates that the product of the transfected E beta b gene associates with the endogenous E alpha chain to form an E alpha dE beta b complex. While expression of E alpha dE beta b is constitutive in A20-2J cells transfected with the E beta b gene, surface expression of E alpha dE beta b is detected in transfected macrophage cells only after treatment of cells with culture supernatants from concanavalin A (Con A)-stimulated T cells. Transfected B lymphoma cells and transfected Con A supernatant-treated macrophage cells have acquired the ability to present antigen to E alpha dE beta b-restricted T-cell hybridomas. The observed inducible expression of the transfected gene in the macrophage host indicates that sequences responsible for regulated expression of the E beta b gene may be associated with the transfected gene. In combination with directed mutagenesis, the system described here provides a means to study (i) E beta b sequences that are important in determining the restriction specificity of the E molecule and (ii) sequences associated with the E beta gene that may be important in the regulation of E beta chain expression.
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