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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。
OBJECTIVE: To investigate the effect of IFN producing killer DC (IKDC) on mice induced by BaF3, a single lymphocyte molecule that co-expressed retinoic acid early transcript 1ε (RAE1ε) and membrane-bound IL-15 ) Functional molecular expression. METHODS: BaF3-mb15 cells expressing membrane type IL-15 and BaF3-RAE cells expressing RAE1ε and BaF3-mb15-RA cells expressing RAE1ε and membrane-type IL-15 were constructed on the basis of the mouse primary B lymphocyte cell line BaF3. RAE cells. Bone marrow-derived mouse mature DCs were induced by cytokines in vitro. Three BaF3-engineered cell lines inactivated by mitomycin were used as stimulator cells and co-cultured with DC respectively. Flow cytometry was used to detect CD11clowB220 + NK1.1 + IKDC ratio and the expression of CD40 and CD80 on DC surface. IKDC was sorted by flow cytometry. Three BaF3-engineered cell lines inactivated by mitomycin C were cultured for 24 hours. Flow cytometry was used to detect the expression of CD40, CD80, CD86, NKG2D, MHC class II molecules, CD107a and FasL expression. RESULTS: Bone marrow-derived mouse mature DCs were successfully obtained. Compared with BaF3-mb15 cells and BaF3-RAE cells, stimulation of BaF3-mb15-RAE cells significantly increased the percentage of IKDC in DCs [(50.0 ± 5.6)% vs (30.3 ± 8.2%, (36.0 ± 4.6% <0.05], and effectively upregulated the expression of CD40 (180.1 ± 28.2 vs 44.7 ± 7.8, 36.0 ± 3.1, P <0.01) and FasL (P <0.05) on the surface of IKDC. Compared with BaF3 and BaF3-mb15 cells, mb15-RAE cells effectively stimulated the up-regulation of CD80 (P <0.05) on the surface of IKDC, while none of the three BaF3-treated cells stimulated the expression of CD40 and CD80 on DCs (P> 0.05) CD86, MHC class II molecules, NKG2D and CD107a expression had no significant effect (P> 0.05). Conclusion: The synergistic effect of RAE-1ε and membrane-type IL-15 can promote the proliferation of IKDC and induce its expression of CD40 and FasL.