恒河猴树突状细胞生物学特性的初步研究

Preliminary Studies on Biological Characteristics of Rhesus Monkey(Macaca mulatta) Dendritic Cells

树突状细胞(dendritic cell,DC)在HIV初次感染和潜伏感染中发挥着关键作用。恒河猴的生物学性质与人类非常接近,所以恒河猴是研究HIV感染的常用动物模型。以恒河猴为实验动物进行DC的研究首先需要对恒河猴DC的生物学性质有初步的了解。本研究从健康恒河猴外周血中分离单个核细胞,用rhGM-CSF、rhIL-4和rhTNF-α等细胞因子组合体外扩增诱导恒河猴DC,成功诱导出具有典型形态特征的DC,所获DC高表达MHCII类分子和CD86共刺激分子,在混合淋巴细胞反应中能有效刺激T细胞增殖,培养至第9天表达CD1a的细胞比例为60.5%±2.0%,表达CD83的比例为43.8%±1.5%,辣根过氧化物酶(horseradish peroxidase,HRP)内吞实验说明诱导的猴DC在第6天吞噬能力最强。本研究为以恒河猴为实验动物进行DC的研究打下了基础。

Dendritic cell （DC） play a crucial role both in the primary HIV infection stage and in the latent HIV infection period. The rhesus macaque, whose biological characteristics are remarkably similar to those of humans, is a good animal model to study the AIDS pathogenesis. The biological characteristics of the rhesus monkey dendritic cells need to be known before the immune function of rhesus monkey dendritic cells in HIV infection will be studied. Rhesus monkey （Macaca mulatta） monocytes were separated from peripheral blood. Monocytes were proliferated and induced to differentiate toward DC by using recombinant human granulocyte- rhGM-CSF, rhlL-4, and impulsed by using inactive SV40 antigen, and promoted for mature DC by using rhTNF-α. The morphocytology of DC was observed under ordinary light microscope and scanning electron microscope. The characteristic dendritic morphology was observed on some DC surface. High expression of HLA-DR and CD86 on rhesus monkey DC were mensurated. Rhesus monkey DC derived from inducement in vitro can cause auto T cell proliferation effectively in mixed lymphocyte reaction （MLR）. At day 9 of cell culture, flow cytometric analysis showed that cells with CDla account for 60.5%±2.0%, and cells with CD83 account for 43.8%±1.5%. DC endocytosis of horseradish peroxidase （HRP） showed that the peak value of endocytic capacity of DC lies at the sixth day of cell culture. This study laid the foundation for the research and development of DC by using rhesus monkey as animal model.