Ginsburg H, Ben-David E, Kinarty A, Rofolovitch M, Amitay M, Chriqui E, Kedar E, Davidson S
Immunology. 1983 Aug;49(4):571-83.
Colonies of cells termed 'giant granular leucocytes' (GGL) displaying natural killer (NK) activity were generated in cell culture. The prominent feature of these cells was the formation of large cytoplasmic pool--the 'theca'--filled up with glycogen. This was demonstrated by the strong positive red staining of the theca with periodic acid Schiff reagent (PAS) which was abolished by prior treatment with amylase. Two different procedures were employed for obtaining colonies of NK-GGL. In the first, mice were injected either with killed Corynebacterium parvum or with killed Bordetella pertusis preparations and their mesenteric lymph-node cells were grown on syngeneic X-irradiated embryonic skin fibroblast monolayers. At the foci of GGL formation the fibroblasts were killed and the cleared areas thus formed were populated by adherent GGL. In the second procedure, supernates from rat or mouse spleen cultures stimulated with concanavalin A (Con A)--Interleukin-2 (IL-2)--were added to cultures of spleen and lymph-node cells prepared from either ordinary or from athymic nude mice. Richest GGL populations developed when rat IL-2 was added to cells of nude mice. Mouse IL-2 was less consistent. With nude mouse cells it stimulated, either mast cells or GGL, or both; rat IL-2 did not stimulate mast-cell differentiation in nude mouse cultures. In contrast, supernates from lymph-node cell cultures prepared from mice infected with Schistosoma mansoni. Mucosal mast cell-stimulating factor (MMSF) stimulated the formation of colonies of mast cells but not GGL. When MMSF was added as late as 23 days, colonies of young mast cells appeared and mast cells progressively increased in number. When rat IL-2 was added to such mature mast-cell cultures on the 30th day, colonies of cytolytic-GGL appeared. These observations indicate that precursors of mast cells and GGL persist in the cultures and preserve their potential to be stimulated by T-cell factors. GGL-NK cells developed on monolayers prepared from whole embryos released substance that displayed morphology and staining characteristic of mucus. Evidence gathered from in-vitro and in-vivo studies links the in-vitro GGL-NK cells to motile cells that inhabit the mucosal epithelium. Based on the observations, a hypothesis on the function of NK cytotoxicity is brought forward. It proposes the replacement of ordinary epithelial cells, which are killed during a proliferative and differentiative response of other cells at the onset of an infection course.
在细胞培养中产生了具有自然杀伤（NK）活性的被称为“巨型颗粒白细胞”（GGL）的细胞集落。这些细胞的显著特征是形成了充满糖原的大细胞质池——“膜鞘”。用高碘酸希夫试剂（PAS）对膜鞘进行强烈的阳性红色染色证明了这一点，而淀粉酶预处理可消除这种染色。采用了两种不同的方法来获得NK - GGL集落。第一种方法是，给小鼠注射灭活的微小棒状杆菌或灭活的百日咳博德特氏菌制剂，然后将它们的肠系膜淋巴结细胞接种在同基因X射线照射的胚胎皮肤成纤维细胞单层上。在GGL形成的部位，成纤维细胞被杀死，由此形成的空白区域被贴壁的GGL占据。在第二种方法中，将用刀豆蛋白A（Con A） - 白细胞介素 - 2（IL - 2）刺激的大鼠或小鼠脾细胞培养上清液添加到从普通小鼠或无胸腺裸鼠制备的脾细胞和淋巴结细胞培养物中。当将大鼠IL - 2添加到裸鼠细胞中时，会形成最丰富的GGL群体。小鼠IL - 2的效果不太稳定。对于裸鼠细胞，它要么刺激肥大细胞，要么刺激GGL，或者两者都刺激；而大鼠IL - 2不会刺激裸鼠培养物中的肥大细胞分化。相比之下，来自感染曼氏血吸虫的小鼠制备的淋巴结细胞培养上清液。黏膜肥大细胞刺激因子（MMSF）刺激肥大细胞集落的形成，但不刺激GGL。当在第23天很晚的时候添加MMSF时，会出现年轻肥大细胞集落，并且肥大细胞数量逐渐增加。当在第30天向这种成熟的肥大细胞培养物中添加大鼠IL - 2时，会出现溶细胞性GGL集落。这些观察结果表明，肥大细胞和GGL的前体细胞存在于培养物中，并保留了被T细胞因子刺激的潜力。在由全胚胎制备的单层上发育的GGL - NK细胞释放出具有黏液形态和染色特征的物质。从体外和体内研究收集的证据将体外GGL - NK细胞与栖息在黏膜上皮的运动细胞联系起来。基于这些观察结果，提出了一个关于NK细胞毒性功能的假说。它提出在感染过程开始时，在其他细胞的增殖和分化反应中被杀死的普通上皮细胞会被替代。
