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同基因突变体B6.C-H-2bm-1(H-2bm-1)对EL4上T细胞受体的血清学反应。

The congenic mutant B6.C-H-2bm-1 (H-2bm-1) serological response to the T-cell receptor on EL4.

作者信息

Saunders P H, McIntyre B W, Lawlor D A, Bridges W J, Allison J P

出版信息

Cell Immunol. 1986 Sep;101(2):586-92. doi: 10.1016/0008-8749(86)90168-1.

Abstract

Antisera reactive with the C57Bl lymphoma EL4 were induced by hyperimmunization of B6.C-H-2bm-1 (H-2bm-1) mice, a congenic mutant strain of C57Bl/6 (B6). Molecular weight determinations of EL4 surface structures detected with the congenic mutant antibodies were accomplished by electrophoretic analyses (one and two dimensions) on sodium dodecyl sulfate-polyacrylamide gels (SDS-PAGE). Analysis of immunoprecipitates in the first dimension under reducing conditions revealed protein bands corresponding to gp 70 and its 33-kDa cleavage fragment, and two-three bands (40-45 kDa) that overlapped with those precipitated from EL4 with AS 8177, an antiserum which detects framework determinants on the T-cell heterodimeric receptor. Further analysis by diagonal electrophoresis confirmed identity of the 40- to 45-kDa proteins precipitated from EL4 with either AS 8177 or H-2bm-1 anti-EL4 sera. Additional diagonal electrophoretic studies showed that the congenic mutant antibodies do not precipitate T-cell receptor molecules from C6VL, an in vitro line derived from the C6XL T-cell lymphoma of C57Bl/ka origin, which was previously shown by some of us to express the heterodimeric T-cell receptor. Together these results demonstrate detection of the heterodimeric T-cell receptor on EL4 with congenic mutant antibodies directed against nonconstant region determinant(s), and indicate a possible linkage between MHC haplotype and the immune response to intraspecies variable regions of the T-cell receptor.

摘要

通过对B6.C-H-2bm-1(H-2bm-1)小鼠(C57Bl/6(B6)的一个同源突变株)进行超免疫,诱导出了与C57Bl淋巴瘤EL4发生反应的抗血清。用同源突变抗体检测到的EL4表面结构的分子量测定,是通过在十二烷基硫酸钠-聚丙烯酰胺凝胶(SDS-PAGE)上进行电泳分析(一维和二维)来完成的。在还原条件下对一维免疫沉淀物的分析揭示了与gp 70及其33-kDa裂解片段相对应的蛋白条带,以及两到三条(40 - 45 kDa)与用AS 8177(一种检测T细胞异二聚体受体上构架决定簇的抗血清)从EL4沉淀出的条带重叠的条带。通过对角线电泳的进一步分析证实了用AS 8177或H-2bm-1抗EL4血清从EL4沉淀出的40至45 kDa蛋白质的同一性。额外的对角线电泳研究表明,同源突变抗体不会从C6VL沉淀T细胞受体分子,C6VL是一种源自C57Bl/ka品系的C6XL T细胞淋巴瘤的体外细胞系,我们中的一些人之前已证明其表达异二聚体T细胞受体。这些结果共同证明了用针对非恒定区决定簇的同源突变抗体检测到了EL4上的异二聚体T细胞受体,并表明MHC单倍型与对T细胞受体种内可变区的免疫反应之间可能存在联系。

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