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目的:探讨视黄酸早期转录因子1ε(retinoic acid early transcript 1ε,RAE-1ε)和膜型IL-15对小鼠NK细胞杀伤功能的影响。方法:前期研究以小鼠原B淋巴细胞株BaF3为基础构建了3株BaF3工程细胞株,即表达膜型IL-15的BaF3/mb15细胞、表达RAE-1ε的BaF3/RAE细胞和同时表达膜型IL-15和RAE-1ε的BaF3/mb15/RAE细胞。将γ射线灭活后的3株BaF3工程细胞株作为刺激细胞,分别刺激小鼠NK细胞。流式细胞术检测刺激后NK细胞表面分子的表达,胞内染色法检测NK细胞穿孔素和颗粒酶B的分泌,流式细胞术检测NK细胞对小鼠淋巴瘤YAC1细胞的杀伤活性。结果:与BaF3/mb15和BaF3/RAE细胞相比,BaF3/mb15/RAE细胞可有效上调NK细胞表面CD25、CD44、FasL和CD107a的表达,但对穿孔素和颗粒酶B的分泌没有明显刺激作用。当效靶比为20:1时,BaF3/mb15、BaF3/RAE和BaF3/mb15/RAE细胞刺激后NK细胞对靶细胞YAC1的杀伤率分别为(39.7±2.9)%、(45.3±2.3)%和(59.0±6.9)%,均高于BaF3组的(28.3±1.5)%(P<0.01),且BaF3/mb15/RAE组高于BaF3/mb15和BaF3/RAE组(P<0.05)。结论:膜型IL-15联合RAE-1ε可促进NK细胞活化并增强NK细胞的杀伤活性。
AIM: To investigate the effects of retinoic acid early transcript 1ε (RAE-1ε) and membrane-type IL-15 on the cytotoxicity of murine NK cells. Methods: Three BaF3-engineered cell lines were constructed on the basis of the mouse original B lymphocyte cell line BaF3, namely BaF3 / mb15 cells expressing membrane type IL-15, BaF3 / RAE cells expressing RAE-1ε, Type IL-15 and RAE-1 epsilon of BaF3 / mb15 / RAE cells. Three BaF3-engineered cell lines inactivated by γ-rays were used as stimulator cells to stimulate mouse NK cells respectively. Flow cytometry was used to detect the expression of NK cell surface molecules after stimulation. The secretion of perforin and granzyme B of NK cells was detected by intracellular staining. The cytotoxicity of NK cells to YAC1 cells was detected by flow cytometry. Results: Compared with BaF3 / mb15 and BaF3 / RAE cells, BaF3 / mb15 / RAE cells could effectively up-regulate the expressions of CD25, CD44, FasL and CD107a on NK cells but not on the secretion of perforin and granzyme B . When the target ratio was 20: 1, the lethal rates of NK cells to target cells YAC1 were (39.7 ± 2.9)% and (45.3 ± 2.3)%, respectively, after being stimulated by BaF3 / mb15, BaF3 / RAE and BaF3 / mb15 / And (59.0 ± 6.9)%, respectively, higher than those in BaF3 group (28.3 ± 1.5)% (P <0.01), and higher in BaF3 / mb15 / RAE group than those in BaF3 / mb15 and BaF3 / RAE group. Conclusion: Membrane-type IL-15 combined with RAE-1ε can promote NK cell activation and enhance NK cell cytotoxicity.