K562及K562-AO2细胞HLA I 类分子及MICA-B表达及其对NK细胞杀伤.docVIP

K562及K562/AO2细胞HLA I 类分子及MICA/B表达及其对NK细胞杀伤 　 作者：梅家转　牛新清　郭坤元　周健　魏红梅 【摘要】 　　本研究探讨K562亲本细胞株及多药耐药细胞株K562/AO2细胞表面HLA I 类分子和MHC I 类链相关分子（MICA/B）的表达及其对NK细胞杀伤活性的影响。用流式细胞仪检测K562和K562/AO2细胞株HLA I类分子和MICA/B的表达情况；LDH释放法测定3例健康人NK细胞在不同效靶比时对K562和K562/AO2细胞的杀伤活性；效靶比10∶1时，用抗HLA I 类分子单克隆抗体（W6/32）和抗MICA/B单克隆抗体（BAMO1）分别封闭MHC Ⅰ类分子和MICA/B，观察NK细胞杀伤K562及K562/AO2细胞活性的变化。结果表明： K562和K562/AO2细胞均不表达HLA I 类分子，K562细胞的MICA/B表达较K562/AO2明显增高（Plt;0.01）。效靶比5∶1、10∶1、20∶1、30∶1时NK细胞杀伤K562和K562/AO2细胞的活性分别为 (29.32±0.12)%、(45.33±0.78 )%、(58.37± 0.87)%、(72.37±0.96)和(12.47±0.91)%、(24.36±1.11)%、(33.29±1.03)%、(53.87 ±1.27)%，各效靶比时NK细胞对K562细胞的杀伤活性较K562/ AO2细胞明显增强（P=0.000）；效靶比10∶1时W6/32不抑制NK细胞杀伤K562和K562 /AO2细胞的活性，BAMO1能明显抑制NK细胞杀伤K562和K562/AO2细胞的活性。结论： MICA/B在K562和K562/AO2细胞表达的差异与NK细胞的杀伤活性有关；NK细胞对K562/AO2细胞杀伤活性降低与HLA I类分子无关；MICA/B在多药耐药肿瘤细胞的低表达导致耐药肿瘤细胞对NK细胞杀伤敏感性下降。 【关键词】 自然杀伤细胞; MHC I 类链相关分子A/B; K562细胞 　　Expression of HLA Class I Molecules and MHC Class I Chainrelated Molecules A/B in K562 and K562/AO2 Cell Lines and Their Effects on Cytotoxicity of NK Cells Abstract The study was aimed to investigate the expression of HLA class I molecules and MHC class I chainrelated molecules A/B (MICA/MICB) in K562 and adriamycin (ADM)resistant K562 cell lines (K562/AO2) and their effect on cytotoxicity of NK cells. Expression of HLA class I molecules and MICA/MICB on the surface of K562 and K562/AO2 cell lines were analyzed by flow cytometry. Cytotoxicity of NK cells (isolated from 3 healthy persons) against K562 and K562/AO2 cells were detected by LDH releasing assay at different effecttotarget cell ratios (E∶T). In blocking experiments, antiMHC class I monoclonal antibody (McAb) (W6/32, a pan antiHLA class I antibody) and antiMHC class I chainrelated molecules McAb (BAMO1, specificly against MICA and MICB) were added to the target cells at E∶T of 10∶1. The results showed that the expression of MHC class I chainrelated molecules on K562 was higher than that on K562/AO2 (P=0.000), and HLA class I molecules were