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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

24卷

期数:

2018年01期

页码:

10-13

栏目:

基础医学

出版日期:

2018-01-20

文章信息/Info

Title:

Specificity and efficiency of YFP labeled NK cells by NKp46-Cre induced YFP reporter system in mice

文章编号:

1006-8147(2018)01-0010-04

作者:

于明航¹; 李 杨¹; 阎 晗¹; 王 瑾¹; 黄 珊¹; 袁顺宗²; 尹 洁¹; 李泽兴¹; 王 玺¹

(1.天津医科大学细胞生物学系，天津300070；2.中国人民解放军307医院头颈肿瘤科, 北京100853)

Author(s):

YU Ming-hang¹; LI Yang¹; YAN Han¹; WANG Jin¹; HUANG Shan¹; YUAN Shun-zong²; YIN Jie¹; LI Ze-xing¹; WANG Xi¹

(1.Department of Cell Biology，Tianjin Medical University, Tianjin 300070,China; 2.Department of Head and Neck Oncology，Chinese People’s Liberation Army 307 Hospital, Beijing 100853, China)

关键词:

免疫器官; 自然杀伤细胞; 转基因小鼠; 荧光报告系统; Cre重组酶

Keywords:

immune organs; natural killer cells; transgenic mice; fluorescence reporting system; crerecombinase

分类号:

R392

DOI:

-

文献标志码:

A

摘要:

目的: 检测NKp46-Cre介导的黄色荧光蛋白（YFP）报告基因系统在小鼠体内自然杀伤细胞的效率以及特异性。方法: 通过ROSA26R-YFP与NKp46-Cre小鼠杂交产生后代并且利用基因型鉴定的方法筛选出双阳性的基因型小鼠，流式细胞术检测小鼠体内免疫器官淋巴结，脾脏以及骨髓中的YFP的表达效率，然后用细胞表面抗体标记脾脏的淋巴细胞，分析淋巴细胞群体中YFP阳性细胞的百分比。结果：选取ROSA26R-YFP与NKp46-Cre小鼠杂交后代中基因型为ROSA26R-YFP（+/+）Cre（+/-）的小鼠为实验组，ROSA26R-YFP（-/-）Cre（-/-）的小鼠为对照组；流式细胞术分析免疫器官（淋巴结、骨髓、脾脏）YFP的阳性细胞的比例分别为0.589%±1.02%、1.89%±1.28%、4.53%±1.54%，对照组YFP的阳性细胞的比例分别为0.008%±0.003%、0.126%±0.08%、0.12%±0.004%；两种小鼠的非免疫器官（心脏）无明显YFP阳性的细胞0.009%±0.0002%、0.03%±0.012%；脾脏淋巴细胞系中各免疫细胞（T细胞、B细胞、NK细胞）YFP阳性百分比为89.4%±1.08%、0.89%±0.56%、0.82%±0.82%。结论：NKp46-cre介导的YFP报告基因系统标记NK细胞具有明显的特异性。

Abstract:

Objective: To detect the efficiency and specificity of YFP labeled NK cells by the NKp46-cremediatedYFPreporting system in mice. Methods: Two-positive genotype mice were screened by ROSA26R-YFPand NKp46-cre mice using genotype identification. Flow cytometry was used to detect the expression efficiency of YFP in the immune organs, spleen and bone marrow of mice, and then the lymphocytes of spleen were labeled by cell surface antibody, and the percentageof YFP positive cells in lymphocyte population was analyzed. Results: The mice with genotype ROSA26R-YFP (+/+)Cre(+/-) from ROSA26R-YFP and Nkp46-cre mice were selected as experimental group, the mice selected ROSA26R-YFP(-/-) Cre(-/-) were the control group and the proportions of the positive cells of the immune organs (lymph node, bone marrow, spleen) were 0.589%±1.02%,1.89%± 1.28%,4.53%±1.54%, the proportion of YFP positive cells in control group were 0.008%±0.003%,0.126%±0.08%,and 0.12%±0.004%;there were no distinct YFP positive cells 0.009%±0.0002%,0.03%±0.012% in the non-immune organs (heart) of the two mice, and the YFP positive percentages of each immune cell(T cell, B cell, NK cell) in Splenic lymphocyte system were 89.4%±1.08%,0.89%±0.56%,and 0.82%±0.82%.Conclusion: Nkp46-Cremediated YFP reported that the gene system labeled NK cells may have distinct specificity.

参考文献/References:

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相似文献/References:

[1]王 玲 综 述,徐燕颖 审 校.自然杀伤细胞表面NKp30受体的研究进展[J].天津医科大学学报,2019,25(05):544.

备注/Memo

备注/Memo:

基金项目 国家重点基础研究发展计划基金资助（2014CB910100）；国家自然科学基金资助项目（31600705，31600693）

作者简介 于明航（1991-），男，硕士在读，研究方向：肿瘤免疫；通信作者：王玺,E-mail：wangxi@tmu.edu.cn。

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更新日期/Last Update: 2018-01-19