恒河猴外周血pDC_1/pDC_2的分离和细胞表现型的研究

Study on the isolation and phenotype of peripheral blood pDC_1 and pDC_2 in rhesus monkey

目的 研究恒河猴外周血树突状细胞 (DC)的亚群 pDC1和pDC2 的分离方法 ,并探讨其细胞表现型。方法 采集健康、SIV阴性的恒河猴外周血 ,用Ficoll Hypaque梯度离心法提取外周血单核细胞 (PBMC) ,利用与恒河猴有交叉反应的人单克隆抗体以及三色流式细胞仪分离 pDC1和pDC2 ,并对其进行细胞表现型的鉴定。结果 首次分离出恒河猴 pDC1和 pDC2 细胞 ;并保持其活性。pDC1细胞表现型为Lineage-,HLA DR+和BDCA1+;pDC2 的细胞表现型为Lineage-,HLA DR+和IL 3Rα+。结论 成功分离和鉴定了与人相似的恒河猴pDC1和pDC2 细胞 ,对今后异种移植排斥反应的研究具有一定的意义。

Objective To study the isolation and phenotype of peripheral blood pDC 1 and pDC 2 in rhesus monkey.Methods Peripheral blood monoclear cells (PBMCs) were isolated from healthy, SIV-negative rhesus monkeys (M.mulatta) using Ficoll-Hypaque density separation. DC precursors were identified and were sorted by 3 color rare-event cytometric flow analysis using human monoclonal antibodies cross reactive with rhesus monkey. Results DC subsets were identified within the lineage- HLA-DR+ fraction of PBMCs and maintained activities. Myeloid DC (pDC 1) showed the phenotype lineage-, HLA-DR+ and BDCA 1+; Lymphoid DC (pDC 2)showed the phenotype lineage-, HLA-DR+ and CD123+ (IL-3Rα+).Conclusions We have identified the rhesus monkey pDC 1 and pDC 2 similar to those from human beings. Identification of pDC 1 and pDC 2 is an important first step towards testing of these important immunomodulatory APC in the therapy of allograft rejection in non-human primates.