Helinski E H, Bielat K L, Ovak G M, Meenaghan M A, Wirth J E, Pauly J L
Department of Molecular Immunology, Roswell Park Memorial Institute, Buffalo, NY 14263.
J Med. 1988;19(5-6):353-68.
Results of studies demonstrating the identification of the interleukin-2 receptor (IL-2R; i.e., anti-Tac) on the membrane ultra-structure of human leukemic T cells with an antibody carrying an electron dense colloidal gold microsphere (e.g., immunogold) that was visualized using a scanning electron microscope (SEM) are reported. Our IL-2R model system employed HTLV-1 retrovirus-infected lymphoblastoid cells of the long-term human leukemic T cell line HUT-102B2. The presence of the IL-2R on these cells was defined using a double antibody procedure that employed as the primary antibody a purified mouse monoclonal anti-Leu-IL-2R antibody (mIgGlk, anti-Tac, CD25), and used as the secondary antibody a goat anti-mouse IgG (gamma-chain specific) antibody that had been covalently bonded to a 40 nm colloidal gold particle. More than 95% of the HUT-102B2 were IL-2R+, and there was a uniform distribution of the IL-2R over the surface of the cells. Corresponding controls were employed in all examinations and included IL-2R- Jurkat human leukemic T cells and isotype identical immunoglobulins. The primary and secondary antibody reagents contained whole human serum and bovine serum albumin, and there was no evidence of the non-specific binding of these antibodies. These studies are the first to demonstrate the presence of a lymphokine receptor on the surface architecture of a cell. We anticipate no difficulty in applying the immunogold/SEM technology to define both normal and malignant cell membrane receptors for other cytokines.
报道了有关研究结果,该研究利用携带电子致密胶体金微球的抗体(如免疫金)在人白血病T细胞膜超微结构上鉴定白细胞介素-2受体(IL-2R;即抗Tac),并通过扫描电子显微镜(SEM)进行观察。我们的IL-2R模型系统采用了感染HTLV-1逆转录病毒的长期人白血病T细胞系HUT-102B2的淋巴母细胞样细胞。这些细胞上IL-2R的存在通过双抗体程序确定,该程序使用纯化的小鼠单克隆抗-Leu-IL-2R抗体(mIgGlk,抗Tac,CD25)作为一抗,使用与40 nm胶体金颗粒共价结合的山羊抗小鼠IgG(γ链特异性)抗体作为二抗。超过95%的HUT-102B2细胞IL-2R呈阳性,且IL-2R在细胞表面均匀分布。所有检查均采用了相应的对照,包括IL-2R阴性的Jurkat人白血病T细胞和同型的免疫球蛋白。一抗和二抗试剂中含有全人血清和牛血清白蛋白,没有证据表明这些抗体会发生非特异性结合。这些研究首次证明了细胞表面结构上存在淋巴因子受体。我们预计在应用免疫金/SEM技术来确定其他细胞因子的正常和恶性细胞膜受体方面不会有困难。