Vav-Cre介导的YFP报告基因系统标记小鼠NK细胞的特异性和效率检测

The Specificity and Efficiency of YFP Labeled NK Cells through Vav-Cre Induced YFP Reporter System in Mice

目的探索Vav-Cre介导的黄色荧光蛋白(YFP)报告基因系统标记小鼠体内自然杀伤(NK)细胞的特异性和效率。方法通过基因型分型筛选ROSA26R-YFP与Vav-Cre小鼠杂交后代中双阳性基因型小鼠,流式细胞术分析免疫器官淋巴结、脾、胸腺以及骨髓细胞的YFP表达效率,通过细胞表面抗体标记淋巴结、脾及骨髓的NK细胞,流式细胞术分析NK细胞群体中YFP阳性细胞百分比。结果 ROSA26R-YFP与Vav-Cre小鼠杂交后代共17只,双阳性基因型小鼠(ROSA26R-YFP(+/-)Vav-Cre)11只;流式细胞术分析淋巴结、脾、胸腺、骨髓细胞中YFP阳性细胞的百分比(%)分别为73.87±1.51、56.07±1.47、86.17±1.74、53.60±3.56,阴性对照组相应器官分别为0.27±0.01、1.33±0.91、0.11±0.01,0.29±0.03,差异有统计学意义(均P<0.01),两种类型小鼠非免疫器官肾中均无明显YFP表达(双阳性鼠0.72%±0.43%,对照鼠0.92%±0.27%,P>0.05);淋巴结、脾和骨髓中NK细胞YFP阳性百分比(%)76.94±0.84、81.66±1.18、88.92±0.77,与阴性对照组比较均明显增高(均P<0.01)。结论 Vav-Cre介导的YFP报告基因系统标记小鼠体内NK细胞具有特异性及高效性。

Objective To explore the specificity and efficiency of YFP labeled natural killer （NK） cells through Vav-Cre induced YFP reporter system in mice. Methods ROSA26R-YFP and Vav-Cre mice were crossed, and their YFP and Cre gene double positive progeny were screened by genotyping. The specificity of YFP in hematopoietic cells from im-mune organs including lymph nodes, spleen, thymus and bone marrow were analyzed by flow cytometry. The percentages of YFP positive cells in NK cells from lymph nodes, spleen and bone marrow were also analyzed by flow cytometry. Results A total of 11 double positive mice （ROSA26R-YFP-（＋/-）VavCre） were obtained in 17 mouse offspring by crossing ROSA26R-YFP mice with Vav-Cre mice. The percentages of YFP positive cells in immune organs including lymph nodes, spleen, thy-mus and bone marrow were 73.87%± 1.51%, 56.07%±1.47%, 86.17%± 1.74%and 53.60%± 3.56%, and there were signifi-cant differences compared with the corresponding negative control cells（0.27%±0.01%, 1.33%±0.91%, 0.11%±0.01%and 0.29%± 0.03%, P〈0.01）. There were no YFP expressions in non-immune organs in double positive mice and in negative control mice （0.72%±0.43%vs 0.92%±0.27%, P〈0.05）. The positive rates of YFP were significantly higher in NK cells in lymph nodes, spleen and bone marrow （76.94%±0.84%、81.66%±1.18%and 88.92%±0.77%） compared with those of control （P〈0.01）. Conclusion YFP marked NK cells through Vav-Cre induced YFP reporter system in mice have high specificity and efficiency.