肺巨噬细胞移植治疗选读.ppt

肺巨噬细胞移植治疗 (PMT) GM-CSF：粒细胞集落刺激生物因子，能够刺激骨髓细胞对肺泡细胞的胞饮以及对面面活性物质的降解。GM-CSF受体即CSFR的缺陷或者GM-CSF抗体均可导致hPAP。 GM-CSF的受体为CSFR,CSFR由α和β链组成， 编码α和β的基因分别为csf2ra和csf2rb。 csf2ra和csf2rb突变 ? CSFRB表达缺陷 ? 阻断GM-CSF信号转导 ? 巨噬细胞对肺泡表面蛋白清除能力下降 ? hPAP 也就是说，hPAP患者肺泡血清中GM-CSF升高，而GM-CSF抗体为阴性 基因治疗简介 基因治疗（gene therapy）是指将外源正常基因导入靶细胞，以纠正或补偿因基因缺陷和异常引起的疾病，以达到治疗目的。也就是将外源基因通过基因转移技术将其插入病人的适当的受体细胞中，使外源基因制造的产物能治疗某种疾病。基因治疗的特异性强，其药物本质为DNA，在体内表达为蛋白质，且基因治疗作用有持续性。基因治疗的策略有基因替代，基因修正，基因抑制，基因增强和免疫调节。 实验方法 实验过程简介 三种小鼠模型1）WT mice(wind type) 2)KO mice(Csf2rb-/-) 3)KO+PMT mice(pulmonary macrophage transplantation) hPAP markers： 1）PAS染色阳性 ORO染色阳性 2）BAL turbiditySP-Dconcentration 3）BAL中，GM-CSF? M-CSF? CSF1? MCR1? mRNA for PU1, PPAR? ABCG1 ↓ 结果 Macrophage characterization after PMT These results demonstrated the cells used for PMT were highly purified, mature macrophages capable of surfactant clearance. Efficacy of PMT of WT macrophages PMT nearly completely resolved the abnormal pulmonary histopathology PMT nearly completely resolved the abnormal pulmonary histopathology Result These results demonstrate PMT had a highly efficacious and durable therapeutic effect on the primary pulmonary and secondary systemic manifestations of hPAP in KO mice. Macrophage engraftment efficiency To determine if WT macrophages had a survival advantage over KO macrophage we measured GM-CSF bioactivity in BAL fluid and found it was detectable in KO but not WT mice WT macrophages had increased survival/proliferation compared to KO macrophages in vitro (Fig. 2a) and accumulated to greater numbers after PMT in KO mice than in WT mice (Fig. 2b and Extended Data Fig. 3d) (d) GFP+ cells in BAL cells from WT or KO mice 2 months after PMT of Lys-MGFP BMDMs. PMT of WT LysMGFP knock-in mouse25 BMDMs into KO mice followed by Ki67 immunostaining revealed PMT-derived cells replicated in vivo (Extended Data Fig. 3e-g). To f