小鼠骨髓树突状细胞制备方法的比较研究

为了比较GM CSF与IL 4 (IL 4DC )以及GM CSF与IL 3(IL 3DC )共刺激培养制备的两种树突状细胞 (DC )的差异 ,采用GM CSF (10 0 0U/ml)和IL 4 (10～ 2 0ng/ml)或IL 3(10～ 2 0ng/ml)从小鼠骨髓培养制备DC ,流式细胞仪分析表型 ,体外饲以颗粒化抗原 (与Latexbead交联的Ovalbumin ,微粒 OVA )或抗原多肽 (Ovalbumin的SL8表位 ,即SIIFEKL )后 ,测定两种DC体外对颗粒化抗原的摄取能力和对抗原多肽的递呈能力 ,以及体内对特异性CTL的诱导能力。结果显示 ,IL 3DC较IL 4DC胞体略大 ,但树突状分支略少 ,表达更高的F4 / 80和更低的NLDC14 5、CD4 0 ,而两者的共刺激分子CD80、CD86和MHCI类分子的表达率无显著差异。体外IL 3DC对颗粒化抗原具有更高的摄取能力和对SIIFEKL多肽有更强的递呈能力 ,但体内对特异性CTL的诱导能力二者无显著差异。研究表明 ,两种方法制备的DC在形态、表型、体外抗原多肽递呈和抗原摄取能力方面存在一定差别 ,但具有相似的体内细胞免疫应答诱导能力

Comparative Study on Preparation of Murine Bone Marrow Dendritic Cells

To compare the morphology,phenotype an d function of dendritic cells grown under GM-CSF and IL-3 (IL-3DC) or GM-CSF and IL-4 (IL-4DC) The dendritic cells were grown from bone marrow of C57BL/6 mice using complete media supplemented with GM-CSF (1 000 U/ml) and IL-3 (10～20 ng/ml) or IL-4 (10～20 ng/ml) Five days later,cells were harvested and phenotyped Their capability of endocytosing FITC labeled particulate antigen (bead-OVA) was assayed with FACScan After pulsed with antigenic peptide (SIIFEKL,an epitope of ovalbumin),their function of in vitro antigen presentation through MH C class I pathway and in vivo CTL response induction was assayed The results showed that compared with IL-4DC,the IL-3DC looked larger with less dendrites They expressed higher F4/80,lower NLDC145,CD40 and MHC class II,but the expressions of CD80,CD86 and MHC class I molecules were similar IL-3DC not only uptook more bead-OVA,but also presented antigenic peptide more efficiently in vitro In vivo studies showed that the two kinds of dendritic cells had similar capability of CTL induction It concludes that there are some differences in morphology, phenotype,antigen uptake and peptide presentation between the two kinds dendritic cells,but they have similar capability of in vivo induction of specific CTL response -