MFI ratio estimation of ZAP-70 in B-CLL by flow cytometry can be improved by considering the isotype-matched antibody signal |
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Authors: | Marquez M-E Deglesne P-A Suarez G Romano E |
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Institution: | Laboratorio de Patología Celular y Molecular, Centro de Medicina Experimental, Instituto Venezolano de Investigaciones Científicas, Caracas, Venezuela. marmarqu@ivic.gob.ve |
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Abstract: | Introduction: The IgVH mutational status of B‐cell chronic lymphocytic leukemia (B‐CLL) is of prognostic value. Expression of ZAP‐70 in B‐CLL is a surrogate marker for IgVH unmutated (UM). As determination of IgVH mutational status involves a methodology currently unavailable for most clinical laboratories, it is important to have available a reliable technique for ZAP‐70 estimation in B‐CLL. Flow cytometry (FC) is a convenient technique for this purpose. However, there is still no adequate way for data analysis, which would prevent the assignment of false positive or negative expression. Methods: We have modified the currently most accepted technique, which uses the ratio of the mean fluorescent index (MFI) of B‐CLL to T cells. The MFI for parallel antibody isotype staining is subtracted from the ZAP‐70 MFI of both B‐CLL and T cells. We validated this technique comparing the results obtained for ZAP‐70 expression by FC with those obtained with quantitative PCR for the same patients. Results: We applied the technique in a series of 53 patients. With this modification, a better correlation between ZAP‐70 expression and IgVH UM was obtained. Conclusions: Thus, the MFI ratio B‐CLL/T cell corrected by isotype is a reliable analysis technique to estimate ZAP‐70 expression in B‐CLL. |
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Keywords: | Chronic lymphocytic leukemia ZAP‐70 flow cytometry IgVH mutational status mean fluorescent index |
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