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〖DK〗∶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。

Objective : To study the effects of membrane-bound IL-15 in combination with retinoic acid early transcript 1ε (RAE1ε) on cytotoxicity of NK cells. Methods: Three derivatives of mouse pro-B lymphocyte cell line BaF3, expressing membrane-bound IL-15 (termed BaF3/mb15), or RAE1ε (termed BaF3/RAE), or both membrane-bound IL-15 and RAE1ε (termed BaF3/mb15/RAE) were respectively constructed in the previous study. Mouse NK cells were isolated and stimulated with irradiated BaF3 derivatives. Phenotype analysis of NK cells was performed by flow cytometry. Meanwhile, perforin and granzyme B expressions were detected in NK cells by intracellular staining; the cytotoxicity of NK cells against YAC1 lymphoma target cells was evaluated by flow cytometry. Results: NK cells stimulated with BaF3-mb15-RAE cells expressed higher levels of CD25, CD44, FasL and CD107a compared to NK cells stimulated with BaF3/mb15 or BaF3/RAE cells. However, BaF3-mb15-RAE cells showed no effects on perforin and granzyme B production of NK cells. When the effector to target ratio was 20∶1, the cytotoxicity rates of BaF3/mb15-stimulated NK cells, BaF3/RAE-stimulated NK cells and BaF3/mb15/RAE-stimulated NK cells against YAC1 cells were (39.7±2.9)%, (45.3±23)% and (59.0±6.9)%, respectively, and significantly increased compared with that of BaF3-stimulated NK cells （\[28.3±1.5\]%, P<0.01). Furthermore, BaF3/mb15/RAE-stimulated NK cells exhibited higher cytotoxicity on YAC1 cells than BaF3/mb15-stimulated or BaF3/RAE-stimulated NK cells (P<0.05). Conclusion: IL-15 combined with RAE1ε promotes the activation and cytotoxicity of NK cells.

国家自然科学基金资助项目（No.81001308，No.30800182，No.31100619）；江苏省自然科学基金资助项目（No. BK2010315）；上海市教委晨光计划资助项目（No.11CG48）