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通过转染小鼠β2-微球蛋白基因挽救Daudi细胞的HLA表达。

Rescue of Daudi cell HLA expression by transfection of the mouse beta 2-microglobulin gene.

作者信息

Seong R H, Clayberger C A, Krensky A M, Parnes J R

机构信息

Department of Medicine, Stanford University Medical Center, California 94305.

出版信息

J Exp Med. 1988 Feb 1;167(2):288-99. doi: 10.1084/jem.167.2.288.

Abstract

The Daudi cell line is a B-lymphoblastoid line derived from a Burkitt lymphoma. Daudi cells lack cell surface expression of class I HLA molecules despite the presence of intracellular class I heavy chains. They have a defect in the gene encoding beta 2-microglobulin (beta 2m), resulting in lack of translatable mRNA for this protein. It has been thought that this deficiency is responsible for the lack of cell surface class I expression. However, data have recently been presented demonstrating that at least one mouse class I heavy chain can be expressed on the cell surface in the absence of beta 2m. These results raised the questions of whether the lack of beta 2m is the only defect in Daudi and whether transfer of this single gene could restore surface class I expression. We found that transfection of the mouse beta 2m gene into Daudi indeed rescued cell surface expression of class I HLA molecules, and that these molecules could be recognized both by monomorphic and allospecific mAbs. CTL clones specific for HLA-B17 or a determinant shared by HLA-B17 and HLA-A2 killed the Daudi cells transfected with the beta 2m gene, but not untransfected Daudi or Daudi transfected with vector alone. Mouse beta 2m on the transfected Daudi cells could exchange with human beta 2m when the cells were incubated in human serum. This exchange did not alter the ability of the cells to be killed by the specific CTLs. These results demonstrate that the lack of beta 2m is the sole reason for lack of surface class I molecules in Daudi cells, and that beta 2m is required for cell surface expression of the specific class I heavy chains of Daudi.

摘要

Daudi细胞系是一种源自伯基特淋巴瘤的B淋巴母细胞系。尽管细胞内存在I类重链,但Daudi细胞缺乏I类HLA分子的细胞表面表达。它们在编码β2微球蛋白(β2m)的基因上存在缺陷,导致该蛋白缺乏可翻译的mRNA。一直以来人们认为这种缺陷是细胞表面缺乏I类表达的原因。然而,最近有数据表明,在缺乏β2m的情况下,至少一种小鼠I类重链能够在细胞表面表达。这些结果引发了以下问题:β2m的缺乏是否是Daudi细胞的唯一缺陷,以及转染这个单一基因是否能够恢复表面I类表达。我们发现,将小鼠β2m基因转染到Daudi细胞中确实挽救了I类HLA分子的细胞表面表达,并且这些分子能够被单克隆和同种特异性单克隆抗体识别。对HLA - B17或HLA - B17与HLA - A2共有的一个决定簇具有特异性的CTL克隆能够杀死转染了β2m基因的Daudi细胞,但不能杀死未转染的Daudi细胞或仅转染了载体的Daudi细胞。当转染的Daudi细胞在人血清中孵育时,其上的小鼠β2m能够与人β2m交换。这种交换并没有改变细胞被特异性CTL杀死的能力。这些结果表明,β2m的缺乏是Daudi细胞表面缺乏I类分子的唯一原因,并表明β2m是Daudi细胞特异性I类重链细胞表面表达所必需的。

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