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目的:研究阿糖胞苷(cytarabine,Ara-C)对白血病细胞共刺激分子B7表达的影响,以及联合双功能抗体anti-CD3/anti-Pgp介导T细胞对耐药白血病细胞的杀伤作用。方法:应用流式细胞术检测白血病细胞株K562和多药耐药白血病细胞株K562/A02细胞经Ara-C刺激不同时间后B7-1、B7-2分子的表达,RT-PCR方法检测B7-1mRNA、B7-2mRNA的表达,MTT法检测经Ara-C刺激的K562和K562/A02细胞对T淋巴细胞增殖的影响。CytoTox96非放射性细胞毒性分析检测Ara-C联合anti-CD3/anti-Pgp微型双功能抗体对人外周血淋巴细胞杀伤K562和K562/A02靶细胞的影响。结果:经Ara-C刺激的K562和K562/A02细胞B7-1、B7-2分子的表达较对照组明显升高;MTT结果显示,经Ara-C刺激的K562和K562/A02细胞能促进T淋巴细胞增殖;Ara-C联合anti-CD3/anti-Pgp双功能抗体在0.39:1～25:1效靶比范围内,随着效靶比的升高,介导T淋巴细胞对K562和K562/A02细胞的杀伤率随之提高,对高表达Pgp的耐药K562/A02细胞尤为明显。结论:Ara-C可上调白血病细胞B7分子的表达,从而增强anti-CD3/anti-Pgp双功能抗体介导的T细胞对靶细胞的体外杀伤作用。
Objective: To investigate the effect of cytarabine (Ara-C) on the expression of costimulatory molecule B7 in leukemia cells and the anti-drug-resistant anti-CD3 / anti-Pgp mediated cytotoxicity of T cells against drug resistant leukemia cells. Methods: The expression of B7-1 and B7-2 in leukemia cell line K562 and multidrug-resistant leukemia cell line K562 / A02 were detected by flow cytometry (FCM) at different time after Ara-C stimulation. The expression of B7- 1 mRNA and B7-2 mRNA were detected by MTT assay. The proliferation of T lymphocytes in K562 and K562 / A02 cells stimulated by Ara-C was detected by MTT assay. CytoTox96 non-radioactive cytotoxicity assay was used to detect the effect of Ara-C combined with anti-CD3 / anti-Pgp micro-diabodies on the killing of K562 and K562 / A02 target cells by human peripheral blood lymphocytes. Results: The expressions of B7-1 and B7-2 in K562 and K562 / A02 cells stimulated by Ara-C were significantly higher than those in control group. MTT results showed that K562 and K562 / A02 cells stimulated by Ara-C could promote T Lymphocyte proliferation was induced by Ara-C combined with anti-CD3 / anti-Pgp bifunctional antibody in the range of 0.39: 1 ~ 25: 1 effective target ratio. / A02 cell killing rate increased, the high expression of Pgp resistant K562 / A02 cells is particularly evident. CONCLUSION: Ara-C can up-regulate the expression of B7 in leukemia cells and enhance the killing effect of anti-CD3 / anti-Pgp bifunctional antibody on target cells in vitro.