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1、细胞因子体外诱导慢性髓细胞性白血病树突状细胞分化的研姓名:2014年6月25日细胞因子体外诱导慢性髓细胞性白血病树突状细胞分化的研宄作者:张玉芳,吴重阳,张连生,柴晔【摘要】木研究探讨干扰素(IFN)治疗慢性粒细胞性白血病(CML)的可能新机制,为临床治疗CML提供新的思路。用Ficoll密度离心法分离骨髓单个核细胞(BMMNC),用不同细胞因子组合体外诱导培养树突状细胞(DC),在倒置显微镜及光镜卜观察DC形态,应用流式细胞术检测其表面标志(CDla,CD83,CD86,HLA-ABC,HLA-DR,CD54),MT
2、T法检测其混合淋巴细胞反应(MLR)能力。结果表明:经不同细胞因子组合所诱导出的DC,其特征性表面分子表达率均高于培养前的DC(PO.Ol);IFN-ot+GM-CSF组DC表达HLA-ABC、HLA-DR明显高于IL-4+GM-CSF培养组(P0.05);而IFN-a+GM-CSF+IL-4组DC的CD86表达率及MLR水平均高于其它细胞因子组合组(P0.05)。干扰素耐药组DC表面分子表达率及MLR水平较新诊断组和干扰素治疗有效组均明显减低(P0.05),但A、B、C各组的CD86表达率及MLR水平在IFN-a+G
3、M-CSF+IL-4因子组合条件下无明显差异。结论:CML骨髓单个核细胞在含有IFN-a的细胞因子组合条件下可分化为具有形态和免疫表型特征的DC,且表达较高的I类和II类分子、共刺激分子、黏附分子及MLR,表明干扰素治疗CML的机理可能与DC有关,这一结果提示通过增强内源性DC功能可能提高CML临床疗效。【关键词】慢性髓细胞性白血病InVitroCytokines-InducedDifferentiationinMononuclearCellDerivedDendriticCellsfromChronicMyeloid
4、Leukemia0.AbstractThepurposeofthisstudywastoinvestigatethemechanismofeffectsofinterferon-alpha(IFN-a)onchronicmyeloidleukemia(CML).Bonemarrowmononuclearcells(BMMNC)wereobtainedfromheparinizedbloodofCMLpatientsbyFicoll-Paquedensitygradientcentrifugation.Theexpres
5、sionsofCDla,CD83,CD86,HLA-ABC,HLA-DRandCD54onDCinducedbyIFN-a+GM-CSF,IFN-a+GM-CSF+IL-4andIL-4+GM-CSFfor7daysinvitrowereassayedbyflowcytometry.Themorphologicfeatureswereobservedbytransmissionandopticalmicroscopy.Themixedlymphocytereactions(MLR)withDCwereevaluated
6、byMTTassay.TheresultsshowedthattheDCculturedindifferentcytokinecombinationsexpressedsignificantlyhigherlevelsofCDla,HLA-ABC,HLA-DR,CD86,CD54,andCD83thanthoseintheprecultured.TheDCgrowingwithIFN-a+GM-CSFexpressedsignificantlyhigherlevelsofHLA-ABC,HLA-DRthanthosei
7、nGM-CSF+IL-4.TheCD86expressionandMLRlevelsinIFN-a+GM-CSF+IL-4increasedsignificantly.TheexpressionrateofDCantigensandMLRintheIFNresistantgroupsignificantlylowerthanthoseinthenewlydiagnosedandtheeffectivelytreatedgroupsafteratleast6monthsofIFN-atreatment(P0.05).Th
8、eDCfromtheIFNresistantgroupdidnotexpresssignificantlyCD86andMLRinIFN-a+GM-CSF+IL-4groupscomparedtothoseinthenewlydiagnosedandIFNeffectivetreatedgroups.Itisconcludedth