Correlation of flow cytometrically determined expression of ZAP-70 using the SBZAP antibody with IgVH mutation status and cytogenetics in 1,229 patients with chronic lymphocytic leukemia

Cytometry B Clin Cytom. 2009 Nov;76(6):385-93. doi: 10.1002/cyto.b.20483.

Abstract

Background: ZAP-70 provides an important prognostic information in chronic lymphocytic leukemia (CLL); however, the most appropriate antibody clone and way of analysis have not yet been defined.

Methods: We determined ZAP-70 expression in 1,229 patients with CLL using the SBZAP clone by applying three different ways of analysis (% positive B-cells, mean fluorescence intensity (MFI) in B-cells, MFI ratio T-cells:B-cells).

Results: ZAP-70 expression was related to somatic hypermutation status of IgVH genes for all three ways of analysis (P < 0.0001 each). The strongest correlation was found for MFI ratio (r = -0.475, P < 0.0001); cases with mutated and unmutated IgVH status significantly differed in their mean MFI ratio (6.33 vs. 3.34, P < 0.0001). ZAP-70 expression was weaker in sole del(13q) (5.44 vs. 4.44, P = 0.001) and stronger in del(11q) (3.31 vs. 5.29, P < 0.0001) and del(6q) (3.29 vs. 5.18, P = 0.005). MFI ratio was significantly related to time to therapy (P = 0.004, RR = 0.79) in 221 evaluable cases. Multivariate analysis proved MFI ratio (P = 0.043, RR = 0.64) and CD38 (P = 0.047, RR = 1.20 per 10%) as only parameters independently related to time to therapy.

Conclusions: Determination of ZAP-70 expression using SBZAP and applying MFI ratio T-cells:B-cells results in significant relations to IgVH mutation status, cytogenetics, and outcome and should be further analyzed and considered for routine application in CLL.

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Antibodies* / genetics
  • Antibodies* / metabolism
  • Chromosome Aberrations
  • Cytogenetics
  • Female
  • Flow Cytometry / methods*
  • Humans
  • Immunoglobulin Heavy Chains / genetics*
  • Immunoglobulin Variable Region / genetics*
  • Kaplan-Meier Estimate
  • Leukemia, Lymphocytic, Chronic, B-Cell* / genetics
  • Leukemia, Lymphocytic, Chronic, B-Cell* / immunology
  • Male
  • Middle Aged
  • Multivariate Analysis
  • Somatic Hypermutation, Immunoglobulin*
  • ZAP-70 Protein-Tyrosine Kinase / metabolism*

Substances

  • Antibodies
  • Immunoglobulin Heavy Chains
  • Immunoglobulin Variable Region
  • ZAP-70 Protein-Tyrosine Kinase
  • ZAP70 protein, human