title: Distinctive patterns of naïve/memory subset distribution and cytokine expression in CD4 T lymphocytes in ZAP-70 B-chronic lymphocytic patients.
abstract: BACKGROUND
  ZAP-70 upregulation in B chronic lymphocytic leukemia (B-CLL) cells is a recognized marker of poor prognosis in these patients; the biological basis of this differential clinical outcome nonetheless remains unknown. ZAP-70 overexpression is considered a surrogate marker of a B-CLL cell subset. To test whether the differential biological characteristics of these patients also include the T helper population, we studied naïve, non-terminated memory (NTEM), terminated memory (TEM) and central memory (CM) cells and cytokine expression by CD4 T lymphocytes from ZAP-70+and ZAP-70-B-CLL patients.
  
  
  METHODS
  Expression of CD3, CD8, CD45RA, CD27, and CD28 antigens and intracytoplasmic cytokine production (IFNγ, IL-2, IL-4, IL-10 and TNFα) were assessed simultaneously by nine-color flow-cytometry in peripheral blood lymphocytes from B-CLL patients. B cell ZAP-70 expression in B-CLL cells was also analyzed by flow cytometry.
  
  
  RESULTS
  Compared to ZAP-70-B-CLL patients, ZAP-70+B-CLL patients showed 1) significant reduction in the naïve T helper subset and expansion of NTEM and TEM subsets, 2) a decrease in the percentage of activated CD4 T lymphocytes expressing IFNγ, TNFα and IL-2, and 3) an increase in the percentage of CD4 T lymphocytes expressing IL-4 or IL-10.
  
  
  CONCLUSIONS
  In conclusion, in early stage B-CLL patients, ZAP-70 upregulation is associated with distinct patterns of activation/differentiation stage subset distribution and of cytokine expression in CD4 T lymphocytes. © 2013 Clinical Cytometry Society.
authors.affiliation: Laboratory of Immune System Diseases and Oncology, Department of Medicine (CNB/CSIC Associated Unit), University of Alcalá, Alcala de Henares, 28871, Madrid, Spain; Hematology Service, Hospital Universitario de la Paz, Madrid, Spain.
authors.forename: Jorge
authors.initials: J
authors.lastname: Monserrat
authors.forename: Miguel
authors.initials: M
authors.lastname: Angel Sánchez
authors.forename: Raquel
authors.initials: R
authors.lastname: de Paz
authors.forename: David
authors.initials: D
authors.lastname: Díaz
authors.forename: Sonia
authors.initials: S
authors.lastname: Mur
authors.forename: Eduardo
authors.initials: E
authors.lastname: Reyes
authors.forename: Alfredo
authors.initials: A
authors.lastname: Prieto
authors.forename: Antonio
authors.initials: A
authors.lastname: de la Hera
authors.forename: Carlos
authors.initials: C
authors.lastname: Martínez-A
authors.forename: Melchor
authors.initials: M
authors.lastname: Alvarez-Mon
keywords: B-CLL; CD4 T lymphocytes; Intracellular cytokine production; ZAP-70; flow cytometry