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目的:探讨表达NKG2D配体视黄酸早期转录因子1ε(retinoic acid early transcript 1ε,RAE1ε)的原B细胞株BaF3诱导产生的髓系抑制性细胞(myeloid-derived suppressor cell,MDSC)的杀伤功能及对NK细胞功能的影响。方法:以小鼠原B细胞株BaF3为基础,构建表达空质粒的BaF3-mock对照细胞和表达RAE1ε的BaF3-RAE1ε细胞。将BaF3-mock和BaF3-RAE1ε细胞分别注射小鼠后诱导产生CD11b~+Gr-1~+MDSC,磁珠分选MDSC后与NK细胞共培养后,流式细胞术检测其对NK细胞表面NKG2D和CD107a表达的影响,ELISA法检测其对NK细胞分泌IFN-γ的影响。将MDSC分别与BaF3-mock和BaF3-RAE1ε细胞共培养后,乳酸脱氢酶释放法检测其对靶细胞BaF3-mock和BaF3-RAE1ε细胞的杀伤作用。结果:与BaF3-mock细胞相比,BaF3-RAE1ε细胞诱导产生的MDSC对NK细胞表面NKG2D和CD107a的表达没有明显影响(P>0.05),对NK细胞分泌IFN-γ的水平也没有显著影响(P>0.05)。与BaF3-mock细胞相比,BaF3-RAE1ε细胞诱导产生的MDSC对靶细胞BaF3-mock和BaF3-RAE1ε的杀伤作用明显增强[(1.99±0.39)%vs(8.63±1.45)%、(5.09±0.67)%vs(17.33±0.41)%,P<0.01]。[(1.20±0.09)%vs(10.31±0.69)%、(5.52±1.64)%vs(18.91±3.04)%,P<0.01]。结论:RAE-1ε增强MDSC对靶细胞的杀伤功能。
OBJECTIVE: To investigate the cytotoxicity of myeloid-derived suppressor cells (MDSCs) induced by BaF3 in naive B cell line expressing NKG2D ligand retinoic acid early transcript 1ε (RAE1ε) Effect on NK cell function. Methods: BaF3-mock control cells expressing empty plasmids and BaF3-RAE1ε cells expressing RAE1ε were constructed based on mouse primary B cell line BaF3. CD11b ~ + Gr-1 ~ + MDSCs were induced after mice were injected with BaF3-mock and BaF3-RAE1ε cells respectively. After being sorted with MDSCs, magnetic beads were co-cultured with NK cells. Flow cytometry was used to detect the expression of NKG2D And CD107a expression. The effect of IFN-γ on NK cells secreting IFN-γ was detected by ELISA. After co-cultured with BaF3-mock and BaF3-RAE1ε cells respectively, MDSCs were tested for cytotoxicity against BaF3-mock and BaF3-RAE1ε cells by lactate dehydrogenase release assay. Results: Compared with BaF3-mock cells, MDSC induced by BaF3-RAE1ε cells had no significant effect on the expression of NKG2D and CD107a on NK cells (P> 0.05), but also had no significant effect on NK cells secreting IFN-γ P> 0.05). Compared with BaF3-mock cells, the killing effect of BaF3-mock and BaF3-RAE1ε induced by BaF3-RAE1ε cells was significantly enhanced [(1.99 ± 0.39)% vs (8.63 ± 1.45)%, (5.09 ± 0.67 )% vs (17.33 ± 0.41)%, P <0.01]. [(1.20 ± 0.09)% vs (10.31 ± 0.69)%, (5.52 ± 1.64)% vs (18.91 ± 3.04)%, P <0.01]. Conclusion: RAE-1ε enhances the cytotoxicity of MDSC on target cells.