畜牧兽医学报 ›› 2016, Vol. 47 ›› Issue (9): 1924-1930.doi: 10.11843/j.issn.0366-6964.2016.09.022

• 基础兽医 • 上一篇    下一篇

不同内吞方式中的关键因子抗体对树突状细胞捕获猪细小病毒样颗粒的影响

申小强,左玉柱*,邸晶美,顾文源,范京惠*   

  1. (河北农业大学动物医学院,保定 071001)
  • 收稿日期:2016-03-09 出版日期:2016-09-23 发布日期:2016-09-29
  • 通讯作者: 左玉柱,男,副教授,硕导,主要从事动物传染病学的教学、科研与社会服务工作,E-mail:zuoyuzhu@163.com|范京惠,女,博士,副教授,硕导,主要从事兽医微生物与免疫学的教学与科研工作,E-mail:jinghui76@163.com
  • 作者简介:申小强(1989-),男,河北邯郸人,硕士生,主要从事动物传染病学研究
  • 基金资助:

    国家自然基金资助项目(31101847)

The Influence of Antibodies against Key Factors of Different Endocytic Pathways on Dendritic Cells Uptake Recombinant Parvovirus-like Particles

SHEN Xiao-qiang,ZUO Yu-zhu*,DI Jing-mei,GU Wen-yuan,FAN Jing-hui*   

  1. (College of Veterinary Medicine,Agricultural University of Hebei,Baoding 071001,China)
  • Received:2016-03-09 Online:2016-09-23 Published:2016-09-29

摘要:

 为了探明猪脾树突状细胞(DC)捕获猪细小病毒样颗粒的方式,通过磁性筛选的方法从猪的脾分离DC,DC与肌动蛋白抗体小窝蛋白caveolin-1抗体、猪IgG受体FcγRI(CD64)的抗体及DEC205抗体分别在37 ℃下作用1 h后,再与荧光素FITC标记的猪细小病毒样颗粒(FITC-PPV-VLPs-E290)在37 ℃下作用1 h,应用流式细胞仪及共聚焦显微镜检测体外DC对FITC-PPV-VLPs-E290的捕获情况。结果显示,经肌动蛋白抗体处理的DC几乎不能捕获FITC-PPV-VLPs-E290,经caveolin-1抗体处理的DC 对PPV-VLPs-E290的捕获效率明显下降。而CD64 及DEC205抗体则对DC的捕获效率无影响,表明DC对PPV-VLPs-E290的捕获依赖于肌动蛋白,与巨胞饮、小窝蛋白介导的内吞方式有关,而与吞噬作用及DEC205依赖的网格蛋白介导的方式无关。

关键词: 病毒样颗粒, 树突状细胞, 捕获方式

Abstract:

 Recombinant parvovirus-like particles (PPV-VLPs) are particulate exogenous antigens that induce strong CTL response.In order to decipher the mechanisms responsible for CTL activation by such exogenous antigen and the probable endocytic pathways in PPV-VLP capture,we analyzed the influence of different antibodies on dendritic cells (DCs) endocytic uptaking recombinant parvovirus-like particles.The results analyzed by FACStar cytometer and confocal microscopy showed that the PPV-VLPs capture efficiency of spleen DCs pretreated with anti-CD64 and anti-DEC205 antibodies was not affected.However,anti-caveolin-1 antibodies treated DCs was severely reduced.DCs pretreated with anti-actin antibodies were barely to uptake PPV-VLPs.These results showed that the uptake of PPV-VLPs seems to be related to macropinocytosis and caveolae-mediated endocytosis in an actin-dependent pathway,not clathrin-coated pits or phagocytosis.

Key words:  parvovirus-like particles, dendritic cells, uptake pathways

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