Human T cell lines differing in phenotype and specificity are reactive with the same anti-idiotypic antibody.
Human T cell lines differing in phenotype and specificity are reactive with the same anti-idiotypic antibody.
复制标题
表型和特异性不同的人类 T 细胞系可与相同的抗独特型抗体发生反应。
DOI：
10.4049/jimmunol.136.2.601
复制
发表时间：
1986
影响因子：
4.4
通讯作者：
H. Spits
中科院分区：
文献类型：
--
作者：
J. Borst;A. Boylston;J. D. de Vries;H. Spits
3D 6是一种选择与T白血病细胞系HPB-ALL上的T细胞抗原受体反应的单克隆抗体，发现与15名正常供体中10名的3 - 13%的外周血T淋巴细胞反应。用同种异体细胞刺激两个供体的外周T细胞，并通过用山羊抗小鼠偶联的人红细胞玫瑰花结3D 6包被的细胞来富集3D 6+细胞，并在含白细胞介素2的培养基中扩增。以这种方式，获得90至100%的3D 6+细胞系，其对同种异体刺激细胞具有细胞毒性。3D 6抗体可以阻断抗原特异性细胞毒性，以及诱导针对在不存在3D 6抗体的情况下不能被杀死的靶细胞的非特异性细胞毒性。3D 6+细胞群含有T4+以及T8+细胞，表明3D 6抗体定义了可能具有各种抗原特异性的T细胞受体群。将一个3D 6+细胞系分成T4+ T8-和T4- T8+群体。采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、等电聚焦和V8蛋白酶肽图等方法，对HPB-ALL和正常细胞系中分离的3D 6反应性T细胞受体进行了生化分析。等电聚焦分析为3D 6抗体检测到许多结构上不同的T细胞受体的想法提供了额外的证据，因为T细胞受体α链在克隆肿瘤系HPB-ALL上去唾液化后电荷均匀，但在3D 6+正常细胞系上电荷保持异质。从T_4 ~+和T_8 ~+3D_6 ~+细胞系中分离的T细胞受体的电荷没有明显差异，但它们的等电聚焦模式不同。V8蛋白酶肽图谱揭示了一方面从HPB-ALL分离的T细胞受体α链与另一方面从正常3D 6+系分离的T细胞受体α链之间的结构差异，而根据该分析，β链在一级结构上没有很大差异。此外，肽图谱表明T4+群体上存在的T细胞受体与T8+群体上存在的T细胞受体之间的一级结构存在差异。
3D6, a monoclonal antibody selected for reactivity with the T cell antigen receptor on the T leukemic cell line HPB-ALL, was found to react with 3 to 13% of peripheral blood T lymphocytes of 10 out of 15 normal donors. Peripheral T cells of two donors were stimulated with allogeneic cells, and the 3D6+ cells were enriched by rosetting 3D6-coated cells with goat anti-mouse-coupled human red blood cells and were expanded in interleukin 2-containing medium. In this way, 90 to 100% 3D6+ cell lines were obtained that were cytotoxic for the allogeneic stimulator cells. 3D6 antibody could block antigen-specific cytotoxicity, as well as induce nonspecific cytotoxicity toward target cells that could not be killed in the absence of the 3D6 antibody. The 3D6+ cell populations contained T4+, as well as T8+ cells, indicating that 3D6 antibody defined a T cell receptor population that might harbor various antigenic specificities. One 3D6+ cell line was separated into T4+ T8- and T4- T8+ populations. 3D6 reactive T cell receptors isolated from HPB-ALL and normal cell lines were analyzed biochemically by means of sodium dodecyl sulfate-polyacrylamide gel electrophoresis, isoelectric focusing, and V8 protease peptide mapping. Isoelectric focusing analysis provided additional evidence for the idea that 3D6 antibody detected a number of structurally distinct T cell receptors, because the T cell receptor alpha-chain was homogeneous in charge after desialation on the clonal tumor line HPB-ALL, but remained heterogeneous in charge on the 3D6+ normal cell lines. No great differences in charge were found between T cell receptors isolated from T4+ and T8+ 3D6+ lines, but their isoelectric focusing patterns were not identical. V8 protease peptide mapping revealed structural differences between the T cell receptor alpha-chain isolated from HPB-ALL on one hand and from the normal 3D6+ lines on the other, whereas the beta-chains did not differ greatly in primary structure according to this analysis. In addition, the peptide mapping suggested differences in primary structure between T cell receptors present on the T4+ population vs those present on the T8+ populations.