K562细胞转染HLA-B分子对NK细胞杀伤功能的影响

Expression of HLA-B Antigen on K562 Cell and Analysis of Its Influence on Cytotoxicity Mediated by NK from PBMC

为了研究不同HLA B分子对NK细胞杀伤活性的影响 ,我们分别构建pcDNA3 HLA B 390 5 2、B 2 70 4、B 5 1 0 2 2基因真核表达载体 ;借助脂质体将各质粒转染入K5 6 2细胞 ,经G4 1 8筛选 ,分别获得阳性表达细胞株 ;并应用LDH法检测转染细胞对不同个体外周血NK细胞杀伤活性的抑制效应。结果显示 :与转染了空质粒的对照组相比 ,外周血NK细胞对K5 6 2 B39的杀伤率无明显影响 ,而对K5 6 2 B2 7,K5 6 2 B5 1的杀伤率降低。当使用针对NK细胞受体KIR3DL1的单抗DX9封闭NK细胞后 ,此抑制效应大部分消失。提示靶细胞表达HLA Bw4分子可明显抑制NK细胞的杀伤效应 ,而表达HLA

At first we constructed the recombinant expression vectors of HLA B*39052,B*2704 and B*51022,respectively;then the vectors were transfected into the human K562 cell line with the help of lipofects The modified cell line with effective expression of respective corresponding protein by the screen of G418 was obtained NK cytotoxicity was observed in peripheral blood mononuclear cells with the LDH method In contrast with empty K562 pcDNA3,K562 B27 and K562 B51 could inhibit NK cytotoxicity significantly,whereas K562 B39 had no inhibition;furthermore this inhibition was specific for KIR3DL1 because the protection could be mainly reversed by blocking the KIR3DL1 molecules with a specific monoclonal antibody (mAb) DX9 So we could get the conclusion that HLA Bw4 and HLA Bw6 molecules had different influences on NK cell cytotoxicity