Comparison of flow cytometric methods for the measurement of ZAP-70 expression in a routine diagnostic laboratory

被引:42
作者
Gibbs, G
Bromidge, T
Howe, D
Hopkins, J
Johnson, S
机构
[1] Taunton & Somerset Hosp, Dept Haematol, Taunton, Somerset, England
[2] Taunton & Somerset Hosp, Leukaemia Res Unit, Taunton, Somerset, England
来源
CLINICAL AND LABORATORY HAEMATOLOGY | 2005年 / 27卷 / 04期
关键词
chronic lymphocytic leukaemia; ZAP-70; prognosis; flow cytometry; immunoglobulin VH genes;
D O I
10.1111/j.1365-2257.2005.00703.x
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Chronic lymphocytic leukaemia (CLL) follows a variable clinical course with patient survival ranging from only a few years despite treatment, to several decades in patients who may never require clinical intervention. Determination of the mutational status of a patient's immunoglobulin heavy chain variable region (Ig V-H) gene has been used to provide prognostic information, but this assay is not available in most laboratories. The discovery of the expression of the protein tyrosine kinase zeta-associated protein (ZAP)-70 in V-H-unmutated CLL cases led to its proposal as a surrogate marker for V-H status. This study investigated the measurement of ZAP-70 expression in CLL using different flow cytometric protocols. Two different antibodies and two different staining methods were compared. The Caltag ZAP-70 antibody and Fix & Perm kit were the easiest to use and were the most sensitive and specific combination, with 91% concordance between ZAP-70 expression and V-H status. Three patients (9%) were discordant (two V-H mutated/ZAP-70 positive, and one V-H unmutated/ZAP-70 negative). No correlation existed between CD38 and either ZAP-70 expression or V-H status. Measurement of ZAP-70 expression using the Caltag antibody/kit combination provides a standardized flow cytometric method that could be introduced into a routine CLL immunophenotyping panel in a clinical diagnostic laboratory.
引用
收藏
页码:258 / 266
页数:9
相关论文
共 27 条
[1]  
BINET JL, 1981, CANCER-AM CANCER SOC, V48, P198, DOI 10.1002/1097-0142(19810701)48:1<198::AID-CNCR2820480131>3.0.CO
[2]  
2-V
[3]   ZAP-70 - A 70 KD PROTEIN-TYROSINE KINASE THAT ASSOCIATES WITH THE TCR ZETA-CHAIN [J].
CHAN, AC ;
IWASHIMA, M ;
TURCK, CW ;
WEISS, A .
CELL, 1992, 71 (04) :649-662
[4]   Expression of ZAP-70 is associated with increased B-cell receptor signaling in chronic lymphocytic leukemia [J].
Chen, LG ;
Widhopf, G ;
Huynh, L ;
Rassenti, L ;
Rai, KR ;
Weiss, A ;
Kipps, TJ .
BLOOD, 2002, 100 (13) :4609-4614
[5]   National Cancer Institute-sponsored Working Group guidelines for chronic lymphocytic leukemia: Revised guidelines for diagnosis and treatment [J].
Cheson, BD ;
Bennett, JM ;
Grever, M ;
Kay, N ;
Keating, MJ ;
OBrien, S ;
Rai, KR .
BLOOD, 1996, 87 (12) :4990-4997
[6]   ZAP-70 expression as a surrogate for immunoglobulin-variable-region mutations in chronic lymphocytic leukemia [J].
Crespo, M ;
Bosch, F ;
Villamor, N ;
Bellosillo, B ;
Colomer, D ;
Rozman, M ;
Marcé, S ;
López-Guillermo, A ;
Campo, E ;
Montserrat, E .
NEW ENGLAND JOURNAL OF MEDICINE, 2003, 348 (18) :1764-1775
[7]   Ig V gene mutation status and CD38 expression as novel prognostic indicators in chronic lymphocytic leukemia [J].
Damle, RN ;
Wasil, T ;
Fais, F ;
Ghiotto, F ;
Valetto, A ;
Allen, SL ;
Buchbinder, A ;
Budman, D ;
Dittmar, K ;
Kolitz, J ;
Lichtman, SM ;
Schulman, P ;
Vinciguerra, VP ;
Rai, KR ;
Ferrarini, M ;
Chiorazzi, N .
BLOOD, 1999, 94 (06) :1840-1847
[8]   ZAP-70 expression is a prognostic factor in chronic lymphocytic leukemia [J].
Dürig, J ;
Nückel, H ;
Cremer, M ;
Führer, A ;
Halfmeyer, K ;
Fandrey, J ;
Möröy, T ;
Klein-Hitpass, L ;
Dührsen, U .
LEUKEMIA, 2003, 17 (12) :2426-2434
[9]   Immunoglobulin V genes and CD38 expression in CLL [J].
Hamblin, BJ ;
Orchard, JA ;
Gardiner, A ;
Oscier, DG ;
Davis, Z ;
Stevenson, FK .
BLOOD, 2000, 95 (07) :2455-2456
[10]   Unmutated Ig VH genes are associated with a more aggressive form of chronic lymphocytic leukemia [J].
Hamblin, TJ ;
Davis, Z ;
Gardiner, A ;
Oscier, DG ;
Stevenson, FK .
BLOOD, 1999, 94 (06) :1848-1854