Ahrens P B, Ankel H
Department of Biochemistry, Medical College of Wisconsin, Milwaukee 53226.
Biochimie. 1988 Nov;70(11):1619-25. doi: 10.1016/0300-9084(88)90297-0.
Chinese hamster ovary cell lines with different types of N-linked oligosaccharides were tested as targets for control and lymphokine treated natural killer (NK) cells. The targets tested were parent cells, Lec1 mutants and Lec4 mutants. Due to an apparent defect in GlcNAc transferase V, Lec4 cells produce complex-type N-linked oligosaccharides devoid of GlcNAc beta(1-6) linked branches. Lec1 cells form only high mannose-type N-linked oligosaccharides because they lack GlcNAc transferase I activity. Lec1 cells are very sensitive to lysis by beta-interferon treated human NK cells, but both parent and Lec4 cells are resistant to NK lysis. The ability to discriminate between parent and Lec1 targets was demonstrated with untreated control effectors as well as those which were pretreated with either beta-interferon, gamma-interferon or interleukin-2. Both control and lymphokine-boosted NK cells exhibit much greater lytic activity against targets having only high mannose-type N-linked oligosaccharides. Five oligosaccharide structures resembling those found on N-linked glycoproteins were tested for their ability to block NK lysis of Lec1 targets. Only the high mannose-type glycopeptide from 7S soybean glycoprotein was inhibitory in the mu molar range. At the same concentration, none of the complex-type oligosaccharides had any effect on lytic activity. The results suggest that a high mannose-type N-linked oligosaccharides is recognized at some step in NK cell-mediated lysis.
将具有不同类型N-连接寡糖的中国仓鼠卵巢细胞系作为对照和经淋巴因子处理的自然杀伤(NK)细胞的靶标进行测试。所测试的靶标为亲本细胞、Lec1突变体和Lec4突变体。由于N-乙酰葡糖胺转移酶V存在明显缺陷,Lec4细胞产生缺乏N-乙酰葡糖胺β(1-6)连接分支的复合型N-连接寡糖。Lec1细胞仅形成高甘露糖型N-连接寡糖,因为它们缺乏N-乙酰葡糖胺转移酶I活性。Lec1细胞对经β-干扰素处理的人NK细胞的裂解非常敏感,但亲本细胞和Lec4细胞均对NK裂解具有抗性。未处理的对照效应细胞以及经β-干扰素、γ-干扰素或白细胞介素-2预处理的效应细胞均表现出区分亲本细胞和Lec1靶标的能力。对照NK细胞和经淋巴因子增强的NK细胞对仅具有高甘露糖型N-连接寡糖的靶标均表现出更强的裂解活性。测试了五种类似于N-连接糖蛋白上发现的寡糖结构阻断Lec1靶标NK裂解的能力。只有来自7S大豆糖蛋白的高甘露糖型糖肽在微摩尔范围内具有抑制作用。在相同浓度下,复合型寡糖均对裂解活性没有任何影响。结果表明,高甘露糖型N-连接寡糖在NK细胞介导的裂解的某个步骤中被识别。