Horwitz J I, Toner M, Tompkins R G, Yarmush M L
Surgical Services, Massachusetts General Hospital, Boston 02114.
Mol Immunol. 1993 Aug;30(11):1041-8. doi: 10.1016/0161-5890(93)90129-y.
The mouse T-cell line CTLL-2 is known to be dependent on interleukin-2 (IL-2) for both growth and viability. These cells possess high affinity IL-2 receptors and have been shown to internalize IL-2 after binding. To determine if internalization of IL-2 is required for the mediation of its signal, IL-2 was covalently coupled to an insoluble matrix via glutaraldehyde cross-linking and CTLL-2 cells were incubated with the immobilized lymphokine matrix. This covalent cross-linking prevents the free lateral diffusion and internalization of the bound IL-2 receptors (IL-2R) while still permitting specific binding between the cells and the immobilized ligands. Although only very limited proliferation was observed during the incubation as assessed by 3H-thymidine incorporation, the viability of the CTLL-2 cells on the immobilized IL-2 matrix was preserved. Cells incubated on the immobilized IL-2 surface could proliferate in response to exogenous soluble IL-2 that was added to the cultures after 36 hours whereas control cultures incubated with an immobilized BSA matrix had died. This indicates that immobilized IL-2 can mediate some of the activity of soluble IL-2 and that internalization of the IL-2 receptor may not be required for at least part of the IL-2 mediated effect.
已知小鼠T细胞系CTLL-2的生长和存活均依赖于白细胞介素-2(IL-2)。这些细胞具有高亲和力的IL-2受体,并且已证明在结合IL-2后会将其内化。为了确定IL-2的内化是否是其信号传导所必需的,通过戊二醛交联将IL-2共价偶联到不溶性基质上,并将CTLL-2细胞与固定化的淋巴因子基质一起孵育。这种共价交联可防止结合的IL-2受体(IL-2R)自由侧向扩散和内化,同时仍允许细胞与固定化配体之间的特异性结合。尽管通过3H-胸苷掺入评估,在孵育过程中仅观察到非常有限的增殖,但CTLL-2细胞在固定化IL-2基质上的活力得以保留。在固定化IL-2表面孵育的细胞在36小时后添加到培养物中的外源性可溶性IL-2刺激下可以增殖,而与固定化牛血清白蛋白(BSA)基质孵育的对照培养物则死亡。这表明固定化的IL-2可以介导可溶性IL-2的某些活性并且至少部分IL-2介导的效应可能不需要IL-2受体的内化。