猪树突状细胞的体外培养及其主要免疫生物学特性研究

In vitro cultivation and characterization of dendritic cells from porcine peripheral blood monocytes

为建立体外诱导、培养猪外周血单核细胞来源树突状细胞(DC)的方法,并对其进行免疫生物学特性鉴定,本实验从猪外周血无菌分离单核细胞—DC前体细胞,以猪源重组GM-CSF和IL-4联合诱导培养,从形态学、表型及功能方面对其进行检测。结果证实,经体外诱导培养的猪DC具有典型的树突状形态;培养6d的未成熟DC表面SLA-II-DR、CD1、CD172a表达的阳性率分别为61.50%、83.10%和24.90%;培养8d的成熟DC表面SLA-II-DR、CD1、CD172a表达的阳性率分别为71.70%、50.10%和19.30%;培养的DC具有吞噬异物的功能,以及刺激同种异体淋巴细胞增殖的能力。本文首次在国内成功地建立了体外培养猪外周血单核细胞来源DC的方法,为进一步研究DC在猪传染性疾病致病机制中的作用奠定了基础。

To investigate the immunological characteristics of induced dendritic cells（DC） from porcine peripheral blood monocytes（PBMs）,the porcine PBMs were isolated and cultured in the presence of granulocyte-macrophage colony-stimulating factor（GM-CSF）,interleukin-4（IL-4） and lipopolysaccharide（LPS） for 8 days.The results showed that the cultured porcine DCs had the typical morphological characterization of dendritic cells.At 6 days＇ cultivation,61.50 % porcine DCs expressed MHC-II,83.10 % expressed CD1 and 24.90 % showed CD172a expression.The matured porcine DCs（8 days cultivation） showed a higher MHC-II expression rate of 71.70 %,but decreased expression rate in CD1（50.10 %） and CD172a（19.3 %）.The cultured DCs exhibited phagocytic ability and stimulated proliferation of naive allergenic T cells.This work was the first in China that the porcine DCs were successfully induced from porcine PBM.