Schäfer H, Burger R
Department of Immunology, Robert Koch Institute, BGA, Berlin, Germany.
Scand J Immunol. 1992 Oct;36(4):587-95. doi: 10.1111/j.1365-3083.1992.tb03227.x.
Rats were immunized with guinea pig T lymphocytes and the spleen cells were fused with cells of a mouse myeloma line. The resulting hybrids were screened for the production of antibodies selectively reacting with guinea pig T cells. The monoclonal antibody (MoAb) H159 was analysed in detail because it bound to T lymphocytes, but not to B lymphocytes or macrophages. Cellular ELISA, cytofluorometry and immunohistology revealed that the antigen detected by H159 is selectively expressed on the majority of peripheral mature T lymphocytes (about 95%). In contrast, it stained only a minor population of thymocytes in FACS analysis. H159 precipitated from NP40 lysate of T cells a protein with a molecular weight of about 90 kDa when separated under non-reducing conditions in SDS-PAGE. Under reducing conditions bands with molecular weights of about 50 kDa were found. After binding to anti-rat Ig coated beads, the MoAb H159 had a mitogenic effect for guinea pig T lymphocytes whereas soluble MoAb H159 in the presence or absence of macrophages was not mitogenic. The cellular expression and molecular characteristics of the H159 antigen together with the mitogenic activity of the antibody for T cells indicate that the MoAb H159 recognizes the guinea pig T-cell receptor for antigen via a constant region determinant.
用豚鼠T淋巴细胞免疫大鼠，然后将脾细胞与小鼠骨髓瘤细胞系的细胞进行融合。对产生的杂交瘤进行筛选，以获得能选择性地与豚鼠T细胞反应的抗体。对单克隆抗体（MoAb）H159进行了详细分析，因为它能与T淋巴细胞结合，但不与B淋巴细胞或巨噬细胞结合。细胞酶联免疫吸附测定、细胞荧光测定和免疫组织学研究表明，H159检测到的抗原选择性地表达于大多数外周成熟T淋巴细胞（约95%）。相比之下，在荧光激活细胞分选分析中，它仅使少数胸腺细胞着色。当在SDS-PAGE中于非还原条件下分离时，H159从T细胞的NP40裂解物中沉淀出一种分子量约为90 kDa的蛋白质。在还原条件下，发现了分子量约为50 kDa的条带。与抗大鼠Ig包被的珠子结合后，单克隆抗体H159对豚鼠T淋巴细胞具有促有丝分裂作用，而在有或没有巨噬细胞存在的情况下，可溶性单克隆抗体H159没有促有丝分裂作用。H159抗原的细胞表达和分子特征以及该抗体对T细胞的促有丝分裂活性表明，单克隆抗体H159通过恒定区决定簇识别豚鼠T细胞抗原受体。
