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Vav-Cre介导的YFP报告基因系统标记小鼠NK细胞的

李丹丹1,孟歆怿2,雷蕾2,尹洁3,王玺2,张玲4   

  1. 1. 天津医科大学基础医学院生理教研室
    2. 天津医科大学基础医学院细胞生物学系
    3. 天津医科大学基础医学研究中心
    4. 天津医科大学基础医学院生理及病生理学系
  • 收稿日期:2013-12-05 修回日期:2014-04-25 出版日期:2014-09-15 发布日期:2014-09-15
  • 通讯作者: 张玲

The specificity and efficiency of YFP labeled natural killer cell through Vav-Cre induced YFP reporter system in mice

  • Received:2013-12-05 Revised:2014-04-25 Published:2014-09-15 Online:2014-09-15

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摘要： 目的 探索Vav-Cre介导的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黄色荧光蛋白（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，与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相比，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。

关键词: Cre重组酶, YFP报告基因, NK细胞, 免疫器官

Abstract: Objective Explore the specificity and efficiency of YFP labeled natural killer cells through Vav-Cre induced YFP reporter system in mice. Methods We crossed ROSA26R-YFP and Vav-Cre mice and screened their YFP and Cre gene double positive progeny by genotyping. Then we analyzed the specificity of YFP in hematopoietic cells from immune organs including lymph nodes, spleen, thymus and bone marrow through flow cytometry. Next we analyzed the percentage of YFP positive cells in natural killer cells from lymph nodes, spleen and bone marrow. Results We obtained 11 double positive mice among 17 offsprings by crossing ROSA26R-YFP mice with Vav-Cre mice. The percentages of YFP positive cells in the immune organs including lymph nodes, spleen, thymus and bone marrow are 73.87%±1.51, 56.07%±1.47, 86.17%±1.74 and 53.60%±3.56, there are significant differences compared with the corresponding negative control cell（0.27%±0.01, 1.33%±0.91, 0.11%±0.01, 0.29%±0.03）,（P<0.01; double positive mice vs CTRL）; nevertheless in non-immune organ kidney, cells were almost YFP negative（double positive mice 0.72%±0.43 vs CTRL0.92%±0.27; P>0.05）. NK cells from lymph nodes, spleen and bone marrow of these double positive mice were 76.94%±0.84, 81.66%±1.18 and 88.92%±0.77 labeled by YFP (P<0.01; double positive mice vs CTRL). Conclusion YFP marked natural killer cells through Vav-Cre induced YFP reporter system in mice has strong specificity and high efficiency.

Key words: Cre recombinase, YFP reporter gene, natural killer cell, immune organ