NKG2D配体RAE1ε对乳腺癌细胞4T1衍生MDSC功能的影响

发布时间：2018-05-06 05:36

本文选题：视黄酸早期转录因子ε + 髓系抑制性细胞　；参考：《中国肿瘤生物治疗杂志》2017年07期

【摘要】：目的:探讨表达小鼠NKG2D配体之一视黄酸早期转录因子1ε(retinoic acid early transcript 1ε,RAE1ε)的原B淋巴细胞Ba F3对乳腺癌细胞株4T1成瘤小鼠来源的髓系抑制性细胞(myeloid-derived suppressor cell,MDSC)功能的影响。方法:以小鼠原B淋巴细胞株Ba F3为基础,构建表达RAE1ε的Ba F3-RAE1ε细胞以及表达空质粒的Ba F3-mock对照细胞。通过4T1原位肿瘤模型诱导产生CD11b+Gr-1+MDSC,将Ba F3-mock细胞和Ba F3-RAE1ε细胞作为刺激细胞,分别与脾MDSC共培养,流式细胞术检测MDSC表面CD40、CD80、F4/80、CD11c的表达和MDSC内活性氧(ROS)的水平;ELISA法检测共培养上清液IL-10、IL-4和IFN-γ的含量;Griess法检测共培养上清液一氮化氮(NO)的浓度。磁珠分选共培养体系中的MDSC,检测裂解后精氨酸酶的活性;另外,将分选后的MDSC与抗CD3/抗CD28抗体活化的脾细胞共培养,CFSE法检测活化的CD3+CD8+T细胞增殖情况。结果:成功获得4T1原位肿瘤模型来源的小鼠脾MDSC。与Ba F3-mock细胞相比,Ba F3-RAE1ε细胞刺激对MDSC分泌IL-4、IFN-γ、IL-10和NO的水平没有明显影响(P0.05);对MDSC表达CD40、CD80、F4/80、CD11c和ROS也没有显著影响(P0.05)。与Ba F3-mock细胞相比,Ba F3-RAE1ε细胞刺激显著提高MDSC的精氨酸酶活性(156.166±1.209 vs 110.135±7.356,P0.01),并明显增强MDSC对CD8+T细胞增殖的抑制作用。结论:RAE-1ε在体外增强4T1成瘤小鼠来源的MDSC的抑制功能。
[Abstract]:Aim: to investigate the effect of Baf3, one of the mouse NKG2D ligands, on the function of myeloid-derived suppressor cell line (MDSCC) derived from mouse breast cancer cell line 4T1, and to investigate the effect of BaF3 on the function of the proto-B lymphocytes expressing retinoic acid early transcript 1 蔚 -RAE1 蔚. Methods: Ba F3-RAE1 蔚 cells expressing RAE1 蔚 and Ba F3-mock control cells expressing empty plasmids were constructed on the basis of mouse B lymphocyte line BaF3. CD11b Gr-1 MDSC was induced by 4T1 in situ tumor model. Ba F3-mock cells and Ba F3-RAE1 蔚 cells were used as stimulating cells and co-cultured with splenic MDSC, respectively. Flow cytometry was used to detect the expression of CD40, CD80, F4 / 80, CD11c on the surface of MDSC and the level of reactive oxygen species (Ros) in MDSC. The levels of IL-10, IL-4 and IFN- 纬 in co-cultured supernatants were detected by Elisa. The activity of arginase was detected by magnetic bead sorting co-culture system, and the proliferation of activated CD3 CD8 T cells was detected by co-culture of separated MDSC and spleen cells activated by anti CD3/ anti CD28 antibody. Results: the mouse spleen derived from 4T1 in situ tumor model was successfully obtained. Compared with Ba F3-mock cells, the stimulation of F3-RAE1 蔚 cells had no significant effect on the levels of IL-10 and no secreted by MDSC, nor on the expression of CD40 / CD80F4 / 80 / CD11c and ROS in MDSC. Compared with Ba F3-mock cells, the arginase activity of MDSC was significantly increased by the stimulation of Ba F3-RAE1 蔚 cells (156.166 卤1.209 vs 110.135 卤7.356p 0.01), and the inhibitory effect of MDSC on the proliferation of CD8 T cells was significantly enhanced. Conclusion in vitro, the inhibitory function of MDSC derived from 4T1 tumorigenic mice is enhanced by 1: RAE-1 蔚.
【作者单位】：扬州大学医学院病原生物学与免疫学教研室;
【基金】：国家自然科学基金资助项目(No.81373130,No.81001308) 江苏省自然科学基金资助项目(No.BK2010315) 扬州大学大学生创新创业训练计划资助项目~~
【分类号】：R737.9

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