| 顾小梅,许晨光,刘思辰,贺丽丽,王 静.PDK1对生发中心的生成、发育及功能的影响[J].现代生物医学进展英文版,2018,(3):401-406. |
| PDK1对生发中心的生成、发育及功能的影响 |
| Effect of PDK1 on the Formation, Development and Function of Germinal Center |
| Received:May 28, 2017 Revised:June 20, 2017 |
| DOI：10.13241/j.cnki.pmb.2018.03.001 |
| 中文关键词: PDK1 生发中心 B细胞 |
| 英文关键词: PDK1 Germinal center B cell |
| 基金项目:高等学校博士学科点专项科研基金项目（20131018988） |
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| 中文摘要: |
| 摘要 目的：研究PDK1对生发中心(GC)的生成、发育及其功能的影响。方法：通过配种小鼠得到在GC B细胞中特异性敲除PDK1的小鼠，然后采用共聚焦显微镜观察小鼠脾脏GC的大小，多色流式细胞分析方法观测PDK1的敲除是否会影响小鼠B细胞的发育，ELISA技术检测PDK1敲除的小鼠经免疫后其体内产生抗体的能力是否受影响，结合平面脂双层抗原呈递系统及全内反射荧光显微镜成像系统(TIRFM)观测PDK1的敲除是否会影响小鼠脾脏IgG细胞P85的磷酸化水平。结果：PDK1的缺失并不会影响小鼠B细胞的发育，细胞群中成熟与不成熟B细胞所占比例均无显著性变化，但在GC B细胞中条件性敲除PDK1会影响小鼠脾脏GC的生成以及T细胞依赖性抗原免疫反应，而且同等条件活化后，GC B细胞中条件性敲除PDK1的小鼠脾脏IgG细胞其胞内分子P85的磷酸化水平显著降低。结论：PDK1对GC的生成、发育及功能都具有重要作用。 |
| 英文摘要: |
| ABSTRACT Objective: To explore the role of PDK1 on the formation, development and function of germinal center(GC). Methods: Pdk1fl/fl mice was crossed with Aicda-Cre mice to get Pdk1fl/fl AicdaCre/+ mice, which conditionally deleted the PDK1 in the B cells of GC. Then multicolor flow cytometry analysis was used to check whether PDK1 deletion would influence the development of GC B cells. Confocal microscope was used to observe the size of GC in SRBC immunized Pdk1fl/fl AicdaCre/+ and Pdk1fl/fl control mice. And ELISA was used to check the ability of producing T cell-dependent antibodies by Pdk1fl/fl AicdaCre/+ mice mice after immunization. Meanwhile, combined planner lipid bilayers antigen presenting system and TIRFM (Total Internal Reflection Fluoresence Microscope) were performed to observe the phosphorylation level of P85, which was the regulatory subunit of PI3K. Results: PDK1 deletion in the B cells of GC didn't influence the development of B cells. However, GC formation as well as T cell-dependent immune responses were reduced. What's more, compare with the Pdk1fl/fl control mice, PI3K activation level of B cells from GC of Pdk1fl/fl AicdaCre/+ mice was impaired. Conclusion: PDK1 played a critical role in the formation, development and function of GC. |
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