Variables Affecting the Quantitation of CD22 in Neoplastic B Cells

被引:52
作者
Jasper, Gregory A. [1 ]
Arun, Indu [1 ]
Venzon, David [2 ]
Kreitman, Robert J. [3 ]
Wayne, Alan S. [4 ]
Yuan, Constance M. [1 ]
Marti, Gerald E. [5 ]
Stetler-Stevenson, Maryalice [1 ]
机构
[1] NCI, Flow Cytometry Unit, Pathol Lab, Ctr Canc Res,NIH, Bethesda, MD 20892 USA
[2] NCI, Biostat & Data Management Sect, Ctr Canc Res, NIH, Bethesda, MD 20892 USA
[3] NCI, Clin Immunotherapy Sect, Mol Biol Lab, Ctr Canc Res,NIH, Bethesda, MD 20892 USA
[4] NCI, Pediat Oncol Branch, Hematol Dis Sect, Ctr Canc Res,NIH, Bethesda, MD 20892 USA
[5] CBER FDA, Flow & Image Cytometry Sect, Cellular & Tissue Therapy Branch, Div Cell & Gene Therapies,Off Cellular Tissues &, Bethesda, MD USA
关键词
CD22; quantitative flow cytometry; QuantiBRITE; ABC; IMMUNOTOXIN RFB4(DSFV)-PE38 BL22; CHRONIC LYMPHOPROLIFERATIVE DISORDERS; CHRONIC LYMPHOCYTIC-LEUKEMIA; FLOW-CYTOMETRY; T-CELLS; EXPRESSION; MALIGNANCIES; TRIAL; STANDARDIZATION; PERFORMANCE;
D O I
10.1002/cyto.b.20567
中图分类号
R446 [实验室诊断]; R-33 [实验医学、医学实验];
学科分类号
1001 ;
摘要
Background: Quantitative flow cytometry (QFCM) is being applied in the clinical flow cytometry laboratory for diagnosis, prognosis, and assessment of patients receiving antibody-based therapy. ABC values and the, effect of technical variables on CD22 quantitation in acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), follicular lymphoma (FCL), hairy cell leukemia (HCL) and normal B cells were studied. Methods: The QuantiBrite System (R) was used to determine the level of CD22 expression (mean antibody bound per cell, ABC) by malignant and normal B cells. The intra-assay variability, number of cells required for precision, effect of delayed processing as well as shipment of peripheral blood specimens (delayed processing and exposure to noncontrolled environments), and the effect of paraformaldehyde fixation on assay results were studied. Results: The QuantiBRITE method of measuring CD22 ABC is precise (median CV 1.6%, 95% confidence interval, 1.2-2.3%) but a threshold of 250 malignant cells is required for reliable CD22 ABC values. Delayed processing and overnight shipment of specimens resulted in significantly different ABC values whereas fixation for up to 12 h had no significant effect. ABC measurements determined that CD22 expression is lower than normal in ALL, CLL, FCL, and MCL but higher than normal in HCL. Conclusions: CD22 expression was atypical in the hematolymphoid malignancies studied and may have diagnostic utility. Technical variables such as cell number analyzed and delayed processing or overnight shipment of specimens impact significantly on the measurement of antigen expression by QFCM in the clinical laboratory. Published 2010 Wiley-Liss, Inc.
引用
收藏
页码:83 / 90
页数:8
相关论文
共 31 条
[1]  
[Anonymous], 2005, H43A2 CLSI
[2]  
[Anonymous], CYTOMETRY B
[3]   Visual Inspection Versus Quantitative Flow Cytometry to Detect Aberrant CD2 Expression in Malignant T Cells [J].
Arun, Indu ;
Wulu, Jacqueline A. ;
Janik, John E. ;
Jasper, Gregory A. ;
Yuan, Constance M. ;
Venzon, David ;
Stetler-Stevenson, Maryalice .
CYTOMETRY PART B-CLINICAL CYTOMETRY, 2010, 78B (03) :169-175
[4]   Quantitative flow cytometry can predict childhood acute lymphoblastic leukaemia presenting with aplasia [J].
Atra, A ;
Abboudi, Z ;
Farahat, N ;
Catovsky, D .
LEUKEMIA & LYMPHOMA, 1997, 27 (1-2) :173-177
[5]   Determination of leucocyte antibody binding capacity (ABC): the need for standardization [J].
Barnett, D ;
Storie, I ;
Wilson, GA ;
Granger, V ;
Reilly, JT .
CLINICAL AND LABORATORY HAEMATOLOGY, 1998, 20 (03) :155-164
[6]  
Cabezudo E, 1999, HAEMATOLOGICA, V84, P413
[7]  
D'Arena G, 2000, AM J HEMATOL, V64, P275
[8]  
Davis KA, 1998, CYTOMETRY, V33, P197, DOI 10.1002/(SICI)1097-0320(19981001)33:2<197::AID-CYTO14>3.0.CO
[9]  
2-P
[10]   Levels of expression of CD52 in normal and leukemic B and T cells:: Correlation with in vivo therapeutic responses to Campath-1H [J].
Ginaldi, L ;
De Martinis, M ;
Matutes, E ;
Farahat, N ;
Morilla, R ;
Dyer, MJS ;
Catovsky, D .
LEUKEMIA RESEARCH, 1998, 22 (02) :185-191