中文摘要：

目的从体外诱导的骨髓来源的树突状细胞（dendritic cells,DC）中分离分泌IFN-γ的杀伤性树突状细胞（IFN-γproducing killer dendritic cells,IKDC）.方法取C57BL/6小鼠骨髓细胞,以小鼠重组GM-CSF和IL-4协同诱导下培养.培养第6天,加LPS刺激.培养第7天,收集细胞,通过流式细胞仪（flowcytometer,FCM）分选出CD11clowB220＋NK1.1＋的细胞,FCM进一步鉴定其表型.并将其与肿瘤细胞系B16F10共培养24 h后,CBA（cytometric bead array）法检测共培养上清中IFN-γ的分泌水平.结果体外培养的骨髓来源的DC中,FCM检测存在CD11clowB220＋NK1.1＋的细胞,进一步鉴定发现其高表达CD49b,低表达MHC II类分子,不表达Gr-1分子;并且与肿瘤细胞B16F10共培养后可分泌大量IFN-γ.结论通过FCM分选的方法可从体外培养的骨髓来源的DC中获得IKDC.

英文摘要：

Objective To isolate the IFN-γ producing killer dendritic cells（IKDC）from bone marrow-derived dendritic cells.Methods Bone marrow cells were extracted from the tibia and femur bones of C57BL/6 mice and cultured in the RPMI 1640 medium with recombinant mouse granulocyte-macrophage colony-stimulating factor（rmGM-CSF）and interleukin（IL）-4 in vitro.At day 6,lipopolysaccharide（LPS）was added to promote DCs＇ maturation and differentiation.At day 7,CD11clowB220＋NK1.1＋ cells were generated from bulk DC by FCM sorting.Expression of CD49b,MHC class II and Gr-1 was assessed using appropriate conjugated mAb.Furthermore,CD11clowB220＋NK1.1＋ cells were stimulated with B16F10 tumor cells and IFN-γ levels were measured in the supematants using CBA kits.Results CD11clowB220＋NK1.1＋cells were found in cultured bone marrow-derived dendritic cells,and these cells with IKDC phenotype highly expressed CD49b but lowly expressed MHC class II,and did not express Gr-1.Furthermore,FCM-sorted CD11clowB220＋NK1.1＋ cells could produce IFN-γ in response to B16F10 tumor cells.Conclusion IKDC can be isolated from bone marrow-derived DC by FCM sorting.