Kohda H, Uede T, Yuasa H, Yamaki T, Osawa H, Diamantstein T, Yodoi J, Kikuchi K
J Immunol. 1986 Sep 1;137(5):1557-63.
Spleen cells obtained from Lewis rats were cultured with 4 micrograms/ml Con A for 96 hr, and activated cells were fused with BW5147 mouse T lymphoma cells. Seven clones obtained by fusion expressed rat IL 2R. The expression of rat IL 2R on those hybrid cells could be up-regulated by IL 2 itself, ATL-derived factor, and TPA and CA++ ionophore. Those IL 2R could be down-regulated by murine MoAb ART-18 that detects rat IL 2R. All hybrid clones produced IL 2 constitutively. IL 2 produced by hybrid cells bound to its receptor and promoted the proliferation of hybrid cells in some clones. Incubation of cells with exogenous IL 2 resulted in the proliferation of hybrid cells, whereas the proliferation of some clones was inhibited by exogenous IL 2, indicating that IL 2 had bifunctional properties on cell growth. Rat IL 2R from 6H2-F9 hybrid cells was studied by both one- and two-dimensional SDS-PAGE with ART-18. The IL 2R derived from 6H2-F9 cells had 72,000 to 77,000 and 40,000 to 48,000 m.w. major components under nonreducing conditions, and had 50,000 to 56,000 m.w. major and 35,000 to 38,000 m.w. minor components under reducing conditions. The 110,000 m.w. component, the third component of IL 2R, was constantly observed in 6H2-F9 hybrid cells.
从Lewis大鼠获得的脾细胞用4微克/毫升刀豆蛋白A培养96小时,然后将活化细胞与BW5147小鼠T淋巴瘤细胞融合。通过融合获得的7个克隆表达大鼠白细胞介素2受体(IL 2R)。这些杂交细胞上大鼠IL 2R的表达可被IL 2自身、成人T细胞白血病衍生因子、佛波酯和钙离子载体上调。那些IL 2R可被检测大鼠IL 2R的鼠单克隆抗体ART - 18下调。所有杂交克隆均组成性产生IL 2。杂交细胞产生的IL 2与其受体结合,并促进一些克隆中杂交细胞的增殖。用外源性IL 2孵育细胞导致杂交细胞增殖,而一些克隆的增殖被外源性IL 2抑制,表明IL 2对细胞生长具有双功能特性。用ART - 18通过一维和二维SDS - PAGE研究了6H2 - F9杂交细胞的大鼠IL 2R。在非还原条件下,源自6H2 - F9细胞的IL 2R具有72,000至77,000和40,000至48,000道尔顿的主要成分,在还原条件下具有50,000至56,000道尔顿的主要成分和35,000至38,000道尔顿的次要成分。在6H2 - F9杂交细胞中持续观察到IL 2R的110,000道尔顿成分,即第三成分。