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[层粘连蛋白参与下自然杀伤细胞对肿瘤细胞的识别与裂解作用]

[Recognition and lysis by natural killers of tumor cells with participation of laminin].

作者信息

Filatova N A, Tiuriaeva I I, Ivanov V A

出版信息

Tsitologiia. 2008;50(1):72-8.

Abstract

According to the data obtained in the present work, the receptor complex of mouse natural killers (NK) includes laminin, antibody to which blocks EK-activity (NKA regardless of the presence of complement. Preincubation of mouse splenocytes with anti-laminin serum led to a decrease in their NKA towards tumor cells-targets (CT), the NKA activity decreasing 2 times with respect to cultivated cells of rat hepatoma HTC, while 10 times - to cultivated cells of human erythroblastosis K562. Pretreatment of aplenocytes with noraml nonimmune serum did not lead to a change of NKA. Quite different was the pattern after the tumor cell preincubation with anti-laminin serum: pretreatment of CT K562 led to a twofold decrease in sensitivity of these cells to NK-lysis, whereas the pretreatment of CT K562, on the contrary, made them twice sensitive to NK-lysis. Electrophoretic separation of protein of CT plasma membranes with subsequent immunoblotting with anti-laminin immune serum revealed the presence oflaminin on HTC cell plasma membrane, which was identified as laminin 8/9 by the mass-spectrometry method, while no laminin was detected on K562 cells. Preincubation of splenocytes with laminin did nor affect NKA with respect to CT K562 and HTC. Pretreatment of CT K562 and HTC with laminin decreased the NKA to zero. The obtained data allow suggesting a doubtless participation of laminin and its receptors in CT cytolysis by NK.

摘要

根据本研究获得的数据,小鼠自然杀伤细胞(NK)的受体复合物包含层粘连蛋白,针对该蛋白的抗体可阻断EK活性(无论是否存在补体,NKA均受影响)。用抗层粘连蛋白血清预孵育小鼠脾细胞会导致其对肿瘤细胞靶标(CT)的NKA降低,相对于大鼠肝癌HTC的培养细胞,NKA活性降低了2倍,而相对于人成红细胞增多症K562的培养细胞则降低了10倍。用正常非免疫血清预处理脾细胞不会导致NKA发生变化。肿瘤细胞与抗层粘连蛋白血清预孵育后的情况则大不相同:对K562 CT进行预处理导致这些细胞对NK裂解的敏感性降低两倍,而对HTC CT进行预处理则相反,使其对NK裂解的敏感性提高了两倍。对CT质膜蛋白进行电泳分离,随后用抗层粘连蛋白免疫血清进行免疫印迹,结果显示HTC细胞质膜上存在层粘连蛋白,通过质谱法鉴定为层粘连蛋白8/9,而在K562细胞上未检测到层粘连蛋白。用层粘连蛋白预孵育脾细胞对K562 CT和HTC的NKA没有影响。用层粘连蛋白预处理K562 CT和HTC可使NKA降至零。所得数据表明层粘连蛋白及其受体无疑参与了NK对CT的细胞溶解过程。

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