Establishment of the Orientation Tube Panel for 8-color Flow Cytometric Immunophenotyping for Patients with Acute Leukemia_West China Medical Journal

Authors：

HUANG Qiaorong ¹ , YU Jianhua ^1,2 , JIANG Nenggang ² , ZENG Tingting ² , JIA Yongqian ² ,  MENG Wentong ¹

1. Laboratory of Stem Cell Biology; 2. Department of Laboratory Medicine; West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P. R. China;

Corresponding author：

MENG Wentong, Email: qiaoqiaohuang@163.com

Keywords：

急性白血病; 八色流式免疫表型分析; 起始管

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

目的　建立急性白血病（AL）患者八色流式免疫表型分析起始管方案。方法　用胞膜CD3（CD3）、CD19、CD10、CD34、CD45、胞浆CD79a（cCD79a）、髓过氧化物酶（MPO)和胞浆CD3（cCD3）等8种抗体建立八色流式染色方案。膜表面抗体直接染色；膜内抗体经固定破膜，再染色后上机检测。将3个血小板减少患者骨髓标本分别进行抗体的单色染色和缺一色染色；最后对17例确诊的AL初发患者标本进行检测。结果　用单色染色来确定染色方案中各抗体的检测电压及荧光补偿；缺一色染色中，阳性细胞群较单色染色变化均＜10%，表明方案中的各抗体相互作用小。17例AL初发患者中，6例急性B淋巴细胞白血病原始细胞均为CD34和CD19阳性，5例cCD79a阳性和4例CD10阳性；4例急性T淋巴细胞白血病患者均为cCD3阳性；6例急性髓细胞白血病均为CD34和MPO阳性；1例B+T混合表型AL患者CD34、cCD3、CD19、cCD79a及CD10均为阳性，MPO和CD3为阴性，此检测方案能够确定各类AL的细胞类型。结论　建立了AL患者八色流式免疫表型分析起始管方案，操作简便快速，适用于临床检测。

Citation： HUANG Qiaorong,YU Jianhua,JIANG Nenggang,ZENG Tingting,JIA Yongqian,MENG Wentong. Establishment of the Orientation Tube Panel for 8-color Flow Cytometric Immunophenotyping for Patients with Acute Leukemia. West China Medical Journal, 2012, 27(4): 555-559. doi: Copy

1.	Borowitz M, Bene MC, Harris NL, et al. Acute leukemias of ambiguous lineage[M]//Swerdlow SH, Campo E, Harris NL, et al. World Health Organization classification of tumors: pathology and genetics of tumors of haematopoietic and lymphoid tissues. Lyon, IARC Press, 2008: 150-155.
2.	牛挺, 刘霆. 造血干细胞移植并发症的防治进展[J]. 华西医学, 2011, 26 (12): 1761-1762.
3.	Brüggemann M, Schrauder A, Raff T, et al. Standardized MRD quantification in European ALL trials: proceedings of the second international symposium on MRD assessment in Kiel, Germany, 18–20 September 2008[J]. Leukemia, 2010, 24 (3): 521-535.
4.	Pedreira CE, Costa ES, Barrena S, et al. Generation of flow cytometry data files with a potentially infinite number of dimensions[J]. Cytometry A, 2008, 73 (9): 834-846.
5.	Weinberg OK, Arber DA. Mixed-phenotype acute leukemia: historical overview and a new definition[J]. Leukemia, 2010, 24(11): 1844-1851.
6.	Craig FE, Foon KA. Flow cytometric immunophenotyping for hematologic neoplasms[J]. Blood, 2008, 111(8): 3941-3967.
7.	Matutes E, Pickl WF, van’t VeerM, et al. Mixed-phenotype acute leukemia: clinical and laboratory features and classification outcome in 100 patients defined according to the WHO 2008[J]. Blood, 2011, 117(11): 3163-3171.
8.	van den Ancker W, Terwijn M, Westers TM, et al. Acute leukemias of ambiguous lineage: diagnostic consequences of the WHO 2008 classification [J]. Leukemia, 2010, 24(7): 1392-1396.
9.	Wood B. 9-color and 10-color flow cytometry in the clinical laboratory[J]. Arch Pathol Lab Med, 2006, 130(5): 680-690.
10.	Preffer F, Dombkowski D. Advances in complex multiparameter flow cytometry technology: applications in stem cell research[J]. Cytometry B Clin Cytom, 2009, 76(5): 295-314.
11.	李丽, 付笑迎, 吴长有. 八色流式细胞术检测人外周血BCG特异性效应型记忆CD4~+T细胞的表型特征[J]. 细胞与分子免疫学杂志，2011, 27(5): 491-493.
12.	李丽, 付笑迎, 吴长有. 多色流式细胞术检测人外周血BCG特异性效应型记忆γδＴ特异性效应型记忆[J]. 免疫学杂志, 2011, 27(5): 369-372.
13.	Mahnke YD, Roederer M. Optimizing a multicolor immunophenotyping assay[J]. Clin Lab Med, 2007, 27(3): 469-487.
14.	Chattopadhyay PK, Hogerkorp CM, Roederer M. A chromatic explosion: the development and future of multiparameter flow cytometry[J]. Immunology, 2008, 125(4): 441-449.

1. 　Borowitz M, Bene MC, Harris NL, et al. Acute leukemias of ambiguous lineage[M]//Swerdlow SH, Campo E, Harris NL, et al. World Health Organization classification of tumors: pathology and genetics of tumors of haematopoietic and lymphoid tissues. Lyon, IARC Press, 2008: 150-155.
2. 　牛挺, 刘霆. 造血干细胞移植并发症的防治进展[J]. 华西医学, 2011, 26 (12): 1761-1762.
3. 　Brüggemann M, Schrauder A, Raff T, et al. Standardized MRD quantification in European ALL trials: proceedings of the second international symposium on MRD assessment in Kiel, Germany, 18–20 September 2008[J]. Leukemia, 2010, 24 (3): 521-535.
4. 　Pedreira CE, Costa ES, Barrena S, et al. Generation of flow cytometry data files with a potentially infinite number of dimensions[J]. Cytometry A, 2008, 73 (9): 834-846.
5. 　Weinberg OK, Arber DA. Mixed-phenotype acute leukemia: historical overview and a new definition[J]. Leukemia, 2010, 24(11): 1844-1851.
6. 　Craig FE, Foon KA. Flow cytometric immunophenotyping for hematologic neoplasms[J]. Blood, 2008, 111(8): 3941-3967.
7. 　Matutes E, Pickl WF, van’t VeerM, et al. Mixed-phenotype acute leukemia: clinical and laboratory features and classification outcome in 100 patients defined according to the WHO 2008[J]. Blood, 2011, 117(11): 3163-3171.
8. 　van den Ancker W, Terwijn M, Westers TM, et al. Acute leukemias of ambiguous lineage: diagnostic consequences of the WHO 2008 classification [J]. Leukemia, 2010, 24(7): 1392-1396.
9. 　Wood B. 9-color and 10-color flow cytometry in the clinical laboratory[J]. Arch Pathol Lab Med, 2006, 130(5): 680-690.
10. 　Preffer F, Dombkowski D. Advances in complex multiparameter flow cytometry technology: applications in stem cell research[J]. Cytometry B Clin Cytom, 2009, 76(5): 295-314.
11. 　李丽, 付笑迎, 吴长有. 八色流式细胞术检测人外周血BCG特异性效应型记忆CD4~+T细胞的表型特征[J]. 细胞与分子免疫学杂志，2011, 27(5): 491-493.
12. 　李丽, 付笑迎, 吴长有. 多色流式细胞术检测人外周血BCG特异性效应型记忆γδＴ特异性效应型记忆[J]. 免疫学杂志, 2011, 27(5): 369-372.
13. 　Mahnke YD, Roederer M. Optimizing a multicolor immunophenotyping assay[J]. Clin Lab Med, 2007, 27(3): 469-487.
14. 　Chattopadhyay PK, Hogerkorp CM, Roederer M. A chromatic explosion: the development and future of multiparameter flow cytometry[J]. Immunology, 2008, 125(4): 441-449.

West China Medical Journal

Establishment of the Orientation Tube Panel for 8-color Flow Cytometric Immunophenotyping for Patients with Acute Leukemia

Abstract Full text Figures/Tables Video References Cited by

Format

Content