Vol 43, No 1 (2005)
Original paper
Published online: 2011-08-19
Modification of immunocytochemical ZAP-70 assay for potential clinical application in B-cell chronic lymphocytic leukemia
Folia Histochem Cytobiol 2005;43(1):19-23.
Abstract
The ZAP-70 protein is a member of the Syk/ZAP protein tyrosine kinase family, normally expressed in T cells and
NK cells but not found in normal, mature B cells. The protein plays a critical role in the initiation of T-cell signaling. Leukemic
cells from patients with B-cell chronic lymphocytic leukemia (B-CLL) that expressed nonmutated immunoglobulin V genes
were found to express levels of ZAP-70 protein that were comparable to those detected in T cells of healthy adults. The ZAP-70
protein expression can be evaluated by flow cytometry and may be used as a prognostic marker in B-CLL patients. We modified
the method of immunocytochemical assessment of ZAP-70 expression. The traditional two-step method with monoclonal
anti-ZAP-70 antibody in the first step followed by FITC-conjugated goat anti-mouse IgG was changed for one-step method
with monoclonal anti-ZAP-70 antibody labeled by Zenon Alexa Fluor 488. The method is simple and fast. The major advantage
of Zenon labeling technique is its compatibility with simultaneous staining of surface antigens. The cells may be earlier
immunostained for CD3, CD19 and/or CD5 to compare of the ZAP-70 kinase expression in B and T cells.
Keywords: B-CLLZAP-70Flow cytometry
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