慢性粒细胞白血病患者外周血单个核细胞诱生树突细胞的体外研究

目的 :体外诱生慢性粒细胞白血病 ( CML)患者外周血单个核细胞 ( PBMNCs)为树突细胞( DCs)。方法 :CML患者 PBMNCs在含 GM- CSF、IL- 4、TNF- α的培养液中培养 1 4d,倒置显微镜、电镜下观察细胞形态 ,流式细胞仪检测细胞的表面标志 ,免疫磁珠 ( MACS)富集所诱生的 DCs,用D- FISH、RT- PCR方法检测 DCs的白血病源性。利用 MTT法检测所诱生 DCs刺激 T细胞增殖能力。结果 :所诱生的细胞高表达 CD80 、CD86、CD1a、HLA- DR,电镜下细胞表面有丰富突起 ,细胞浆内有丰富线粒体 ,D- FISH、RT- PCR检测所诱生的 DCs具有 bcr/abl融合基因。CML DCs具有刺激 T细胞增殖能力。结论 :体外利用 CML患者 PBMNCs成功诱导生成了来源于 CML细胞的 DCs( CML DCs)。

Study on Induction of Dendritic Cells Derived from Peripheral Blood Mononuclear Cells of Patients with Chronic Myelocytic Leukemia in Vitro

Objective:To induce dendritic cell (DCs) from peripheral blood mononuclear cells of patients with chronicl leukemia (CML). Methods:PBMNCs of patients with CML were induced by GM CSF, IL 4 , TNF α in vitro for 14 days. The induced cells were detected for morphological and phenotypic properties by inversed microscope, electro microscope and FACS. The induced cells were detected for bcr/abl fusion gene by FISH, RT PCR. The ablility of induced cells to stimulate proliferation of allogenic T lymphocyte was tested by MTT test. Results:PBMNCs of patients with CML induced by IL 4, GM CSF, TNF α expressed high level of HLA DR, CD 86 , CD 1a . Electron microscope revealed that the induced cell surface had many irregular cytoplasmatic processes. DCs enriched by MACS had bcr/abl fusion gene that observed by FISH, RT PCR. DCs derived from CML cell (CML DCs) had the ability to stimulate profilication of allogenic T cells. Conclusion: DCs with bcr/abl fusion gene could be induced successfully from CML PBMNCs.