中文摘要：

目的：探讨单独或共同表达视黄酸早期转录因子1ε（retinoic acid early transcript 1ε,RAE1ε）和膜型IL-15的B淋巴细胞BaF3对小鼠产生IFN的杀伤性DC（IFN producing killer DC,IKDC）功能分子表达的影响。方法：以小鼠原B淋巴细胞株BaF3为基础,分别构建表达膜型IL-15的BaF3-mb15细胞和表达RAE1ε的BaF3-RAE细胞及同时表达膜型IL-15和RAE1ε的BaF3-mb15-RAE细胞。通过体外细胞因子诱导产生骨髓来源的小鼠成熟DC,将丝裂霉素灭活后的3株BaF3工程细胞株作为刺激细胞,分别与DC共培养,流式细胞术检测其对DC中CD11clowB220＋NK1.1＋IKDC比例和DC表面CD40、CD80表达的影响。流式细胞术分选DC中的IKDC,将丝裂霉素灭活后的3株BaF3工程细胞株与其共培养24 h,流式细胞术检测共培养对IKDC表面CD40、CD80、CD86、NKG2D、MHCⅡ类分子、CD107a和FasL表达的影响。结果：成功获得骨髓来源的小鼠成熟DC。与BaF3-mb15细胞和BaF3-RAE细胞相比,BaF3-mb15-RAE细胞刺激显著提高DC中IKDC比例[（50.0±5.6）%vs（30.3±8.2）%、（36.0±4.6）%,均P〈0.05],并且有效刺激IKDC表面CD40（180.1±28.2 vs 44.7±7.8、36.0±3.1,P〈0.01）和FasL（P〈0.05）表达上调;与BaF3细胞和BaF3-mb15细胞相比,BaF3-mb15-RAE细胞有效刺激IKDC表面CD80（P〈0.05）表达上调,而3株BaF3工程细胞刺激均不影响DC表面CD40、CD80的表达（P〉0.05）;同时,3株BaF3工程细胞对IKDC表面CD86、MHCⅡ类分子、NKG2D和CD107a的表达也没有显著影响（P〉0.05）。结论：RAE-1ε和膜型IL-15协同作用可促进IKDC增殖并诱导其高表达CD40和FasL。

英文摘要：

Objective： To study the effects of murine bone marrow-derived pro-B（ BaF3） cells overexpressing membrane-bound form of IL-15（ mbIL-15） and retinoic acid early transcript 1ε（ RAE1ε）,either each alone or two in combination,on the expression of functional molecules in murine interferon producing killer dendritic cells（ IKDCs） in vitro.Methods： Clones of BaF3 cells overexpressing mbIL-15,RAE1ε or both mbIL-15 and RAE1ε were established and inactivated by mitomycin. Mouse bone marrow mononuclear cells were isolated and induced into mature DCs by GM-CSF and IL-4. Mature DCs were co-cultured with the three mitomycin inactivated-BaF3 derivatives respectively. At 24 and 72 hours after co-culture,the percentage of CD11clowB220＋NK1. 1＋IKDCs and levels of CD40 and CD80 in DCs and levels of CD40,CD80,CD86,NKG2 D,MHC class Ⅱ molecule,CD107 a and FasL in IKDCs were determined by flow cytometry.Results： BaF3 cells overexpressing both mbIL-15 and RAE were more effective than BaF3 cells overexpressing mbIL-15 or RAE alone to stimulate the DC to IKDC conversion（ [50. 0 ± 5. 6]% vs [30. 3 ± 8. 2]%,[36. 0 ± 4. 6]%,P 0. 05）.IKDCs co-cultured with BaF3 cells overexpressing mbIL-15 and RAE had higher levels of CD40（ 180. 1 ± 28. 2 vs 44. 7 ±7. 8,36. 0 ± 3. 1,P 0. 01） and FasL（ P 0. 05） as compared to IKDCs co-cultured with mbIL-15- or RAE-overexpressing BaF3 cells. IKDCs co-cultured with BaF3 cells overexpressing mbIL-15 and RAE had higher levels of CD80（ P 0. 05） as compared to IKDCs co-cultured with BaF3 cells or mbIL-15-overexpressing BaF3 cells. In contrast,co-culture with BaF3 derivatives showed no effects either on CD40 and CD80 expression in DCs（ P 0. 05） or on CD86,MHC classⅡ molecule,NKG2 D and CD107 a expression in IKDCs. Conclusion： IL-15 combined with RAE1ε may promote the proliferation and CD40 and FasL expression in IKDCs.