摘要
目的研究靶向肝脏过表达细胞因子IL-15对机体免疫系统的影响。方法构建IL-15重组表达质粒pLIVE-IL-15,采用高压注射的方式尾静脉注射C57BL/6小鼠,ELISA检测小鼠血清中IL-15的表达水平,流式检测小鼠脾脏和肝脏淋巴细胞亚群变化。结果成功构建pLIVE-IL-15质粒,pLIVE-IL-15质粒高压注射小鼠后血清中能检测到IL-15持续表达。与pLIVE-EGFP对照组相比,pLIVE-IL-15质粒注射鼠天然免疫系统NK细胞和NKT细胞以及获得性免疫系统的CD4+T细胞和CD8+T细胞数目均显著增加(P<0.05)。体内过表达IL-15促进NK细胞高表达CD69和IFN-γ,而NKT细胞活化状态改变不明显,IL-15能显著诱导CD8+T细胞高分泌IFN-γ(P<0.05),而对CD4+T细胞IFN-γ分泌影响较小。结论 pLIVE-IL-15靶向肝脏后能在体内持续表达,并且能调节天然免疫系统和获得性免疫系统反应。
Objective To determine the effect of persistent over-expression of IL-15 in the liver on immune system. Methods Recombinant plasmid pLIVE-IL-15 encoding IL-15 was constructed, and the plas- mid of 10 p,g was then delivered into the liver of C57BL/6 mice by hydrodynamic tail vein injection. Serum IL-15 was detected by EL1SA in 1 and 3 d, and 1,2, 3 and 4 weeks after injection. Spleen and liver lympho- cytes were analyzed by flow cytometry. Results p-LIVE-IL-15 plasmid was successfully constructed, and hydrodynamic injection of the plasmid pLIVE-IL-15 resulted in a persistent expression of IL-15 in the serum, reaching the peak in 1 week and keeping in a high level till in 4 weeks after injection. Compared with pLIVEEGFP group, pLIVE-IL-15 treatment significantly increased the numbers of innate immune cells （ NK cells and NKT cells） and acquired immune cells （ CIM ＋ T cells and CD8 T cells） significantly （ P 〈 0. 05 ）. In vivo over-expression of IL-15 enhanced the expression of CD69 and IFN-y in NK cells, but had no effect on the activation status of NKT ceils. IL-15 dramatically stimulated the production of IFN-y by CD8 T cells （P 〈 0. 05）, and mildly affected the intracellular expression of IFN-y by CD4 T cells. Conclusion Hydrodynamic injection of the plasmid pL1VE-IL-15 results in a persistent and high-level expression of IL-15 in mouse serum, and modulation in the immune responses of innate and acquired immune cells.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2013年第16期1692-1697,共6页
Journal of Third Military Medical University
基金
国家自然科学基金面上项目(81171560)~~
