title: Comparative analysis of flow cytometric techniques in assessment of ZAP-70 expression in relation to IgVH mutational status in chronic lymphocytic leukemia.
abstract: We compared 1 subjective and 5 objective flow cytometric methods to evaluate zeta-associated protein (ZAP-70) expression in relation to immunoglobulin heavy-chain variable-region (IgVH) gene mutational status in 154 samples from 125 patients with chronic lymphocytic leukemia (CLL). ZAP-70 expression determined by all methods used correlated with IgVH gene mutational status, but none of them demonstrated high concordance rates. Of the objective methods, ZAP-70 staining determined as a ratio of molecules of equivalent soluble fluorochrome intensity in CLL cells to that in normal B cells (ZAP-70+ staining in IgVH germline cases, 59%; ZAP-70- in IgVH mutated cases, 75%) or T cells (ZAP-70+ in IgVH germline cases, 66%; ZAP-70- in IgVH mutated cases, 57%) provides the best combination for assigning ZAP-70+ status to IgVH germline and ZAP-70- status to IgVH mutated cases. The subjective method based on ZAP-70 expression in natural killer/T cells gave a similar result, but reproducibility between laboratories may be difficult. Further studies on ZAP-70 expression in relation to clinical parameters may address whether ZAP-70 is an independent prognostic marker for CLL.
authors.affiliation: Department of Pathology, John H Stroger, Jr Hospital of Cook County, Chicago, IL, USA.
authors.forename: Yi-Hua
authors.initials: YH
authors.lastname: Chen
authors.forename: LoAnn C
authors.initials: LC
authors.lastname: Peterson
authors.forename: David
authors.initials: D
authors.lastname: Dittmann
authors.forename: Andrew
authors.initials: A
authors.lastname: Evens
authors.forename: Steven
authors.initials: S
authors.lastname: Rosen
authors.forename: Adela
authors.initials: A
authors.lastname: Khoong
authors.forename: T Vincent
authors.initials: TV
authors.lastname: Shankey
authors.forename: Meryl
authors.initials: M
authors.lastname: Forman
authors.forename: Rohit
authors.initials: R
authors.lastname: Gupta
authors.forename: Charles L
authors.initials: CL
authors.lastname: Goolsby
mesh_terms: D004252:DNA Mutational Analysis; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D015973:Gene Expression Regulation, Leukemic / Q000235:genetics; D050438:Genes, Immunoglobulin Heavy Chain / Q000235:genetics; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D009154:Mutation; D051746:ZAP-70 Protein-Tyrosine Kinase / Q000096:biosynthesis*
chemical_list: D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D051746:ZAP-70 Protein-Tyrosine Kinase