急性髓系白血病患者骨髓中树突状细胞亚群的定量分析

Quantitative Analysis of Dendritic Cell Subsets in Bone Marrow of Patients with Acute Myeloid Leukemia

本研究探讨初诊急性髓系白血病(AML)患者骨髓中树突状细胞(dendritic cells,DC)亚群数量的变化特点。采用直接免疫荧光标记法,对30例健康志愿者和77例初诊AML患者的骨髓进行髓样DC(myeloidDC,mDC)和浆细胞样DC(plasmacytoid DC,pDC)标记,用流式细胞仪进行检测,并用4色与传统3色两种设门方法进行分析。结果表明:与正常人骨髓中mDC与pDC的数量相比,4色设门方法分析显示,在77例初诊AML患者中:mDC数量减低、正常与增高的患者分别占74.0%(57例)、6.5%(5例)和19.5%(15例),pDC数量减低、正常与增高的患者分别占90.9%(70例)、7.8%(6例)与1.3%(1例);3色设门方法分析显示,mDC数量减低、正常与增高的患者分别占58.4%(45例)、19.5%(15例)和22.1%(17例);pDC数量减低、正常与增高的患者分别占46.8%(36例)、26.0%(20例)与27.3%(21例)。两种设门方法所得结果,有统计学差异(p<0.05)。在AML-M2、M3与M4/M5几个亚型中,4色设门方法分析显示:mDC数量减低患者所占比例分别为81.4%、100%与42.1%;pDC数量减低患者所占比例分别为88.4%、100%与89.5%。M4/M5型与M2、M3型相比,mDC数量正常或增高的患者较多,差异有统计学意义(p<0.05);pDC在各亚型中的数量变化无统计学差异(p>0.05)。结论:4色设门方法比传统3色设门方法更适用于初诊AML患者骨髓中mDC与pDC数量的研究;大部分初诊AML患者表现为mDC与pDC数量的减低;M4/M5型与M2、M3型相比,mDC数量正常或增高的患者较多,而pDC在各亚型之间未见明显差别。

In order to study the quantity of dendritic cell （DC） subsets of bone marrow in patients with acute myeloid leukemia（ AML）, the bone marrow aspirate were collected from 77 newly diagnozed AML patients and from 30 healthy persons. The quantity of DC subsets （ myeloid dendritic cells, mDC and plasmacytoid dendritic cells, pDC ） were detected by flow cytometry and analysed by 3-color and 4-color cytometric gate. Based on the conventional 3-color panel, mDC were identified by Lin- HLA-DR ＋ CD11 c^ ＋ and pDC were identified by Lin- HLA-DR ^＋ CD123 ^＋ . Based on the 4-color panel, mDC were identified by Lin- HLA-DR＋ CDllc＋ BDCA-1 ＋ and pDC were identified by Lin- HLA- DR^ ＋ CD123^ ＋ BDCA-2 ^＋. The results showed that a reduction of mDC was found in 74.0% （57/77） and 58.4% （45/ 77 ） patients, a reduction of pDC was found in 90.9 % （ 70/77 ） and 46.8 % （ 36/77 ） patients respectively by 3-color and 4-color cytometric analysis. Meanwhile an expansion of mDC was showed in 19.5% （15/77） and 22. 1% （17/77） patients, an expansion of pDC was showed in 1.3 % （ 1/77 ） and 27.3 % （21/77） patients respectively by 3-color and 4- color cytometric analysis. In subtypes of AML-M2, AML-M3 or AML-M4/5, 81.4%, 100% and 42.1% patients showed mDC decrease and 88. 4%, 100% and 89. 5% patients showed pDC decrease respectively by 4-color cytometric analysis. It is concluded that the 4-color cytometric gate is better method for detection of mDC and pDC from bone marrow of newly diagnozed AML patients as compared with 3-color cytometric gate, the majority of AML patients showed reduction of mDC and pDC. The percentages of patients with mDC normal or mDC increase in AML-M4/5 subtypes are more than that in AML-M2/3 subtypes, while the pDC does not show difference between AML subtypes.