CD_3AK细胞的体外诱导及免疫生物学特征的实验研究

An experimental study on inducement in vitro andimmunobiological features of CD_3AK

目的 :研究CD3AK细胞的体外诱导方法及其免疫生物学特征。方法 :采用抗单克隆抗体(anti CD3McAb)和基因重组人白细胞介素2(rIL 2)、植物血凝素(PHA)共同诱导人外周血单核细胞(PBMCS)制备CD3AK细胞 ,应用APAAP法、吉姆萨染色法分析CD3AK细胞的表型、核型等免疫生物学特征。结果 :微量的an ti CD3McAb(终浓度30～5000ng/ml)辅以少量的rIL 2(100u/ml)和PHA(100μg/ml)共同培养 ,就能诱导和大量扩增CD3AK细胞 ,细胞扩增倍数随着Anti_CD3McAb的使用浓度提高到5000ng/ml时 ,CD3AK细胞扩增倍数可达800倍以上 ,显著高于LAK细胞的扩增倍数 ,细胞常呈集落生长。CD3AK细胞的表型为CD3、CD4、CD8 的CD3AK细胞比率分别为(72±14) %、(29±4) %、(47±23) %。结论 :CD3AK细胞是一类以CD3、CD4、CD8 细胞为主的异质细胞群 ,并具有淋巴母胞样特征 ,CD3AK细胞为正常二倍体细胞。CD3AK细胞易于体外诱导和大量扩增 ,是一种很有应用前景的肿瘤免疫细胞。

Objective:To investigate the inducement method in vitro and immunobiological features of CD3AK.Method:We adopted anti CD3 monoclonal antibody(anti CD3McAb)、recombinant human interleukin 2 (rIL 2) and phytohemagglutinin(PHA) to induce jointly human peripheral blood mononucleus cells(PBMCs) and used APAAP method、Giemsa's Stain method to assay the immunobiological features of CD3AK,such as phenotype and nucleiotype.Results:CD3AK could be induced and proliferated by trace anti CD3McAb(end concentration 30～500ng/ml) plus rIL 2(100u/ml) and PHA(100μg/ml),and cell proliferation multiple increased along with anti CD3McAb concentration to be enhanced.When anti CD3McAb rose to 500ng/ml,proliferation multiple of CD3AK could reach more than 800 times,and markedly higher than that of LAK.CD3AK grows in colony;the cell ratio with CD3、CD4、CD8 of CD3AK were separately (72±14)%、(29±4)%、(47±23)%,it shows that CD3AK is a group of heterogeneous cells with lymphoblast like features.Conclusion:CD3AK is easily to be induced and proliferate in vitro and it is a kind of promising tumor immunocompetence cell.