Abstract
This paper shows a protocol for the detection of ZAP-70 expression in B-CLL (B cell chronic lymphocytic leukemia) tumor cells by common immunohistochemical methods. The study was conducted on bone marrow trephine biopsies from 62 B-CLL patients at the time of diagnosis. Immunohistochemical reactions based on peroxidase and alkaline phosphatase reactions were used, as well as double immunofluorescent labeling for ZAP-70 detection as an indirect marker of mutated and unmutated CLL. Clinical relevance of the ZAP-70 expression detection method was assessed using χ 2 test between ZAP-70 positivity data and other known prognostic factors, i.e., clinical and cytogenetics data. ZAP-70 was detected in 13 out of 62 patients. Statistically significant results were obtained for ZAP-70 positive cases and known indicators of worse prognosis. Immunohistochemical analysis supported by double immunfluorescent labeling, as shown here, is an easy and reliable technique for the detection of ZAP-70 expression in B-CLL tumor cells applicable in every hematopathology laboratory.
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Acknowledgments
This study was funded by the Croatian Ministry of Science Education and Sports, project 108-1081873-1891, ‘Prognostic value of FOXP1 and FOXP3 in B lymphoproliferative disorders’. The authors would like to thank Marija Milkovic Perisa, MD, for her help in gathering clinical data.
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Korać, P., Ajduković, R., Kardum Paro, M.M. et al. Immunohistochemical analysis of ZAP-70 expression in chronic lymphocytic leukemia. J Mol Hist 40, 81–86 (2009). https://doi.org/10.1007/s10735-009-9213-6
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DOI: https://doi.org/10.1007/s10735-009-9213-6