Matsuda T, Hirano T, Kishimoto T
Institute for Molecular and Cellular Biology, Osaka University, Japan.
Eur J Immunol. 1988 Jun;18(6):951-6. doi: 10.1002/eji.1830180618.
Murine hybridomas producing monoclonal antibodies (mAb) specific to human interleukin 6 (IL 6/BSF-2) were established. One of these hybridomas (MH60.BSF2) was found to be dependent on IL 6 for its in vitro growth. None of the known biological factors tested, such as recombinant (r) human (Hu) IL 1 alpha, rHuIL 1 beta, rHuIL 2, rHuIL 3, rHuIL 4, rHu interferon (IFN)-gamma, HuIFN-beta, rHuG-CSF, or recombinant murine (Mu) IL 3, MuIL 4, rMuIL 5, could induce the in vitro growth of MH60.BSF2 cells. The half-maximum tritiated thymidine uptake by MH60.BSF2 cells could be achieved by picogram amounts of rIL 6, making this hybridoma clone an indicator cell for specific and sensitive detection of the IL 6 activity in test samples. The MH166.BSF2 clone was found to produce IgG1,chi type mAb (alpha BSF2-166) capable of neutralizing IL 6 activity. The other clone, MH60.BSF2, produced IgM,chi type mAb (alpha BSF2-60) unable to neutralize IL 6 activity. Both mAb specifically reacted with IL 6 as demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting analysis. An enzyme-linked immunosorbent assay (ELISA) utilizing alpha BSF2-166 and rabbit anti-IL 6 antibodies was established which could detect as low as 50 pg/ml of IL 6. Since both the ELISA and MH60.BSF2 hybridoma could detect small amounts of IL 6 in biological fluids, they constitute powerful tools in exploring the presence or the role of IL 6 in various immunological disorders.
建立了产生针对人白细胞介素6（IL-6/BSF-2）的单克隆抗体（mAb）的小鼠杂交瘤。发现其中一种杂交瘤（MH60.BSF2）在体外生长依赖于IL-6。所测试的已知生物因子，如重组（r）人（Hu）IL-1α、rHuIL-1β、rHuIL-2、rHuIL-3、rHuIL-4、rHu干扰素（IFN）-γ、HuIFN-β、rHuG-CSF，或重组小鼠（Mu）IL-3、MuIL-4、rMuIL-5，均不能诱导MH60.BSF2细胞的体外生长。皮克量的rIL-6就能使MH60.BSF2细胞摄取氚化胸腺嘧啶达到半数最大值，这使得该杂交瘤克隆成为检测测试样品中IL-6活性的特异且灵敏的指示细胞。发现MH166.BSF2克隆产生能够中和IL-6活性的IgG1、κ型mAb（αBSF2-166）。另一个克隆MH60.BSF2产生不能中和IL-6活性的IgM、κ型mAb（αBSF2-60）。如十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和蛋白质印迹分析所示，两种mAb均能与IL-6特异性反应。利用αBSF2-166和兔抗IL-6抗体建立了酶联免疫吸附测定（ELISA），其可检测低至50 pg/ml的IL-6。由于ELISA和MH60.BSF2杂交瘤都能检测生物体液中的少量IL-6，它们构成了探索IL-6在各种免疫紊乱中的存在或作用的有力工具。
