Maghazachi A A, Vujanovic N L, Herberman R B, Hiserodt J C
Pittsburgh Cancer Institute, Department of Pathology, Pittsburgh 15213.
J Immunol. 1988 Apr 15;140(8):2846-52.
The developmental relationships among large agranular lymphocytes (LAL) large granular lymphocytes (LGL) and the activation of these cells into lymphokine-activated killer (LAK) cells by rIL-2 was investigated. Highly enriched populations of LAL were isolated from Fischer 344 spleen cells by a combination of nylon-wool filtration (to remove B cells and macrophages), treatment with a pan T cell antibody plus complement (to remove T cells) and incubation in L-leucine methyl ester (to remove LGL). The resultant cells were highly enriched in morphologically identifiable LAL which expressed asialo GM1 and partially expressed the OX8 surface marker. The enriched LAL did not contain detectable NK cytotoxic activity, did not express pan T cell (OX19), Ia, Ig, or laminin surface markers and contained less than 0.2% LGL. Incubation of LAL in a low dose of rIL-2 (100 U/ml) induced the generation of LGL having NK activity within 24 h of culture. Longer culture periods (48 h) resulted in a continued increase in the percentage of LGL and higher levels of NK activity. However, with this low dose of rIL-2, little or no LAK activity (i.e., reactivity against NK-resistant target cells) was generated. With a high dose of rIL-2 (500 U/ml), LAL responded by first generating LGL with NK activity (within 24 h), with subsequent generation of LAK activity by 48 h. Evidence that the development of granular lymphocytes from LAL was responsible first for NK activity and then LAK activity was demonstrated by depletion of the generated granular NK or LAK effector cells by second treatments with L-leucine methyl ester. Concomitant with the induction of LGL with NK or LAK activity, rIL-2 also caused LGL to proliferate and expand four- to five-fold in 48 h. This occurred in the presence of high or low dose rIL-2. These results indicate that LAL are the precursors of LGL/NK cells, that LAL, LGL/NK cells and LAK cells appear to represent sequential developmental or activation stages and that LAL may comprise major source of LAK progenitors in lymphoid populations having few LGL or mature active NK cells.
研究了大颗粒淋巴细胞（LAL）、大颗粒淋巴细胞（LGL）之间的发育关系，以及重组白细胞介素-2（rIL-2）将这些细胞激活为淋巴因子激活的杀伤细胞（LAK细胞）的过程。通过尼龙毛过滤（去除B细胞和巨噬细胞）、用泛T细胞抗体加补体处理（去除T细胞）以及在L-亮氨酸甲酯中孵育（去除LGL）相结合的方法，从Fischer 344脾细胞中分离出高度富集的LAL群体。所得细胞在形态上高度富集可识别的LAL，其表达无唾液酸GM1并部分表达OX8表面标志物。富集的LAL不含有可检测到的NK细胞毒性活性，不表达泛T细胞（OX19）、Ia、Ig或层粘连蛋白表面标志物，且LGL含量低于0.2%。将LAL在低剂量rIL-2（100 U/ml）中孵育，在培养24小时内诱导产生具有NK活性的LGL。更长的培养时间（48小时）导致LGL百分比持续增加和NK活性水平升高。然而，使用这种低剂量的rIL-2，几乎不产生或不产生LAK活性（即对NK抗性靶细胞的反应性）。使用高剂量rIL-2（500 U/ml）时，LAL的反应先是在24小时内产生具有NK活性的LGL，随后在48小时产生LAK活性。通过用L-亮氨酸甲酯进行第二次处理耗尽产生的颗粒状NK或LAK效应细胞，证明了从LAL发育而来的颗粒淋巴细胞首先负责NK活性然后是LAK活性。与具有NK或LAK活性的LGL诱导同时发生的是，rIL-2还导致LGL在48小时内增殖并扩增4至5倍。这在高剂量或低剂量rIL-2存在的情况下都会发生。这些结果表明LAL是LGL/NK细胞的前体，LAL、LGL/NK细胞和LAK细胞似乎代表了连续的发育或激活阶段，并且LAL可能是在LGL或成熟活性NK细胞较少的淋巴群体中LAK祖细胞的主要来源。
