ラットインターロイキソ2レセプター発現T細胞ハイブリドーマの確立とその調節機構の解析
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Spleen cells obtained from Lewis rats were cultured with 4μg/ml Con A for 96 hr and the activated cells were fused with BW5147, mouse T lymphoma cells. Three clones were obtained by fusion and expressed the rat interleukin 2 receptor (IL-2R). The expression of rat IL-2R on those hybrid cells could be up-regulated by supernatants of Con A activated spleen cells (condition medium; CM), IL-2 absorbed CM and IL-2 itself. IL-2R on hybrid cells could be down-regulated by murine monoclonal antibody, ART-18 that detects rat IL-2R. T cell hybridomas could express the rat IL-2R with high (Kd=109.6 pM) and low (Kd=5.35 nM) affinity for recombinant IL-2. Incubation of hybrid clones with IL-2 resulted in the proliferation of hybrid cells, whereas the proliferation of some clones was inhibited by IL-2, indicating that IL-2 had bifunctional properties on cell growth. All hybrid clones produced IL-2 constitutively. IL-2 produced by hybrid cells were bound to its receptor and promoted the proliferation of hybrid cells in some clones. The increased expression of IL-2R and secretion of IL-2 induced by the synergistic action of Ca++ inonophores and TPA indicated the importance of the Ca++ dependent protein kinase C in the T cell activation. Rat IL-2R from Con A activated rat T cells and T cell hybridomas were studied by both one-and two-dimensional SDS-PAGE with ART-18. The IL-2R derived from Con A activated rat T cells had 72-77 kd minor and 40-48 kd major components under non-reducing conditions and had 50-56 kd major and 35-38 kd minor components under reducing conditions. The 35-38 kd minor components in reducing conditions were derived from 40-48 kd major components in non-reducing conditions. The IL-2R derived from T cell hybridomas had 110 kd, 72-77 kd and 45-48 kd components under non-reducing conditions and had 110 kd, 50-56 kd and 35-38 kd components under reducing conditions. The 110 kd components, the third component of rat IL-2R was constantly observed in T cell ?hybridomas. The rat IL-2R derived from Con A activated normal rat T cells was found to have a pI between 5.0-5.6 and 6.0-6.4, whereas the rat IL-2R derived from T cell hybridomas was found to have a pI between 6.0-6.4. T cell hybridomas expressed an aberrant IL-2R on their cell surface. The aberrant IL-2R had a molecular weight of 40 kd under reducing conditions and a pI between 5.6-6.2.
- 札幌医科大学の論文
- 1986-10-01
札幌医科大学 | 論文
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