ZAP-70 expression in B-cell chronic lymphocytic leukemia: Evaluation by external (isotypic) or internal (T/NK cells) controls and correlation with IgVH mutations

Antonella Zucchetto, Riccardo Bomben, Michele Dal Bo, Paola Nanni, Pietro Bulian, Francesca Maria Rossi, Maria Ilaria Del Principe, Simone Santini, Giovanni Del Poeta, Massimo Degan, Valter Gattei

Research output: Contribution to journalArticle

Abstract

Background: Expression of T cell specific zeta-associated protein 70 (ZAP-70) by B-cell chronic lymphocytic leukemia (B-CLL) cells, as investigated by flow cytometry, has both prognostic relevance and predictive power as surrogate for immunoglobulin heavy chain variable region (IgVH) mutations, although a standardization of the cytometric protocol is still lacking. Methods: Flow cytometric analyses for ZAP-70 were performed in peripheral blood samples from 145 B-CLL (124 with IgVH mutations) by a standard three-color protocol. Identification of ZAP-70+ cell population was based on an external negative control, i.e., the isotypic control (ISO method) or an internal positive control, i.e., the population of residual normal T/NK cells (TNK method). A comparison between these two approaches was performed. Results: While 86/145 cases were concordant as for ZAP-70 expression according to the two methods (ISO+TNK+ or ISO -TNK-), 59/145 cases had discordant ZAP-70 expression, mainly (56/59) showing a ISO+TNK- profile. These latter cases express higher levels of ZAP-70 in their normal T cell component. Moreover, discordant ISO+TNK- cases had a IgVH gene mutation profile similar to that of concordantly positive cases and different from ZAP-70 concordantly negative B-CLL. Conclusion: Analysis of ZAP-70 expression by B-CLL cells by using the ISO method allows to overcome the variability in the expression of ZAP-70 by residual T cells and yields a better correlation with IgVH gene mutations. A receiver operating characteristic analysis suggests to employ a higher cut-off than the commonly used 20%. A parallel evaluation of the prognostic value of ZAP-70 expression, as determined according to the ISO and TNK methods, is still needed.

Original languageEnglish
Pages (from-to)284-292
Number of pages9
JournalCytometry Part B - Clinical Cytometry
Volume70
Issue number4
DOIs
Publication statusPublished - Jul 15 2006

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B-Cell Chronic Lymphocytic Leukemia
Natural Killer Cells
Mutation
Proteins
T-Lymphocytes
Immunoglobulin Heavy Chains
Cellular Structures
ROC Curve
Population
Genes
Flow Cytometry
Color

Keywords

  • B-CLL
  • Propuostic factors
  • ZAP-70

ASJC Scopus subject areas

  • Hematology
  • Cell Biology
  • Pathology and Forensic Medicine
  • Biophysics
  • Endocrinology

Cite this

ZAP-70 expression in B-cell chronic lymphocytic leukemia : Evaluation by external (isotypic) or internal (T/NK cells) controls and correlation with IgVH mutations. / Zucchetto, Antonella; Bomben, Riccardo; Dal Bo, Michele; Nanni, Paola; Bulian, Pietro; Rossi, Francesca Maria; Del Principe, Maria Ilaria; Santini, Simone; Del Poeta, Giovanni; Degan, Massimo; Gattei, Valter.

In: Cytometry Part B - Clinical Cytometry, Vol. 70, No. 4, 15.07.2006, p. 284-292.

Research output: Contribution to journalArticle

Zucchetto, Antonella ; Bomben, Riccardo ; Dal Bo, Michele ; Nanni, Paola ; Bulian, Pietro ; Rossi, Francesca Maria ; Del Principe, Maria Ilaria ; Santini, Simone ; Del Poeta, Giovanni ; Degan, Massimo ; Gattei, Valter. / ZAP-70 expression in B-cell chronic lymphocytic leukemia : Evaluation by external (isotypic) or internal (T/NK cells) controls and correlation with IgVH mutations. In: Cytometry Part B - Clinical Cytometry. 2006 ; Vol. 70, No. 4. pp. 284-292.
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abstract = "Background: Expression of T cell specific zeta-associated protein 70 (ZAP-70) by B-cell chronic lymphocytic leukemia (B-CLL) cells, as investigated by flow cytometry, has both prognostic relevance and predictive power as surrogate for immunoglobulin heavy chain variable region (IgVH) mutations, although a standardization of the cytometric protocol is still lacking. Methods: Flow cytometric analyses for ZAP-70 were performed in peripheral blood samples from 145 B-CLL (124 with IgVH mutations) by a standard three-color protocol. Identification of ZAP-70+ cell population was based on an external negative control, i.e., the isotypic control (ISO method) or an internal positive control, i.e., the population of residual normal T/NK cells (TNK method). A comparison between these two approaches was performed. Results: While 86/145 cases were concordant as for ZAP-70 expression according to the two methods (ISO+TNK+ or ISO -TNK-), 59/145 cases had discordant ZAP-70 expression, mainly (56/59) showing a ISO+TNK- profile. These latter cases express higher levels of ZAP-70 in their normal T cell component. Moreover, discordant ISO+TNK- cases had a IgVH gene mutation profile similar to that of concordantly positive cases and different from ZAP-70 concordantly negative B-CLL. Conclusion: Analysis of ZAP-70 expression by B-CLL cells by using the ISO method allows to overcome the variability in the expression of ZAP-70 by residual T cells and yields a better correlation with IgVH gene mutations. A receiver operating characteristic analysis suggests to employ a higher cut-off than the commonly used 20{\%}. A parallel evaluation of the prognostic value of ZAP-70 expression, as determined according to the ISO and TNK methods, is still needed.",
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T1 - ZAP-70 expression in B-cell chronic lymphocytic leukemia

T2 - Evaluation by external (isotypic) or internal (T/NK cells) controls and correlation with IgVH mutations

AU - Zucchetto, Antonella

AU - Bomben, Riccardo

AU - Dal Bo, Michele

AU - Nanni, Paola

AU - Bulian, Pietro

AU - Rossi, Francesca Maria

AU - Del Principe, Maria Ilaria

AU - Santini, Simone

AU - Del Poeta, Giovanni

AU - Degan, Massimo

AU - Gattei, Valter

PY - 2006/7/15

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N2 - Background: Expression of T cell specific zeta-associated protein 70 (ZAP-70) by B-cell chronic lymphocytic leukemia (B-CLL) cells, as investigated by flow cytometry, has both prognostic relevance and predictive power as surrogate for immunoglobulin heavy chain variable region (IgVH) mutations, although a standardization of the cytometric protocol is still lacking. Methods: Flow cytometric analyses for ZAP-70 were performed in peripheral blood samples from 145 B-CLL (124 with IgVH mutations) by a standard three-color protocol. Identification of ZAP-70+ cell population was based on an external negative control, i.e., the isotypic control (ISO method) or an internal positive control, i.e., the population of residual normal T/NK cells (TNK method). A comparison between these two approaches was performed. Results: While 86/145 cases were concordant as for ZAP-70 expression according to the two methods (ISO+TNK+ or ISO -TNK-), 59/145 cases had discordant ZAP-70 expression, mainly (56/59) showing a ISO+TNK- profile. These latter cases express higher levels of ZAP-70 in their normal T cell component. Moreover, discordant ISO+TNK- cases had a IgVH gene mutation profile similar to that of concordantly positive cases and different from ZAP-70 concordantly negative B-CLL. Conclusion: Analysis of ZAP-70 expression by B-CLL cells by using the ISO method allows to overcome the variability in the expression of ZAP-70 by residual T cells and yields a better correlation with IgVH gene mutations. A receiver operating characteristic analysis suggests to employ a higher cut-off than the commonly used 20%. A parallel evaluation of the prognostic value of ZAP-70 expression, as determined according to the ISO and TNK methods, is still needed.

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