Table 1.
Characteristics of the patients at CLL diagnosis.
Figure 1.
Flow cytometric analysis of Th17 cells.
The dot plots show representative data from healthy control subjects, illustrating the analysis method for identification of CD4/CD3+/IL-17A+ cells (Th17). An acquisition gate was established basing on FSC and SSC that included mononuclear cells. A region, R1, was drawn around the lymphocytes (A). Next, the R1 gated events were analyzed for CD3 PE-Cy5 and CD4 FITC staining and positive cells (CD4+/CD3+) were gated (region R2) (B). The final dot plots CD4FITC versus mouse IgG1 PE (C) and CD4FITC versus IL-17A PE (D) were established by combined gating of events using R1 and R2. The number in the upper right quadrant on the dot plot D represents the percentage of CD4+/CD3+/IL-17A+ (Th17) cells.
Figure 2.
Plasma levels of IL-17A and percentage of Th17 cells in CLL patients and HVs.
(A) The IL-17A concentration in PB from CLL patients an healthy volunteers (HVs) (39.35 pg/ml vs. 14.94 pg/ml, p = 0.0018). (B) The percentage of CD4+/CD3+/IL-17A+ (Th17) cells in CLL and HVs (8.76% vs. 3.02%, p = 0.035).
Figure 3.
Plasma IL-17A levels and percentage of Th17 cells in CLL patients in different disease stages.
(A) Plasma levels of IL-17A in CLL patients with Rai stages: 0 (median: 44.63 pg/ml), I–II (median: 35.33 pg/ml) and III–IV (median: 15.23 pg/ml). (B) Median percentage of Th17 cells in CLL patients with Rai stage 0 (median, 12.11%) in comparison to those in Rai stages I–II (median, 8.11%) and III–IV (median, 5.15%).
Figure 4.
Plasma levels of IL-17A and percentage of Th17 cells in CLL patients analyzed by adverse prognostic factors.
(A) Median plasma level of IL-17A in ZAP-70-negative patients in comparison to ZAP-70-positive patients (45.36 pg/ml vs. 26.11 pg/ml, p = 0.0062) (B). Median plasma level of IL-17A in CD38-negative patients and CD38-positive patients (39.73 pg/ml vs. 29.25 pg/ml, p = 0.033). (C) The percentage of CD4+/CD3+/IL-17A+ cells in ZAP-70-negative patients compared with ZAP-70-positive patients (4.85% vs. 2.67%, p = 0.016). (D). Median percentage of CD4+/CD3+ cells with intracellular IL-17A expression in CD38-negative and CD38-positive patients (4.46% vs. 2.87%, p = 0.028). (E,F) Plasma levels of IL-17 and percentage of Th17 cells in CLL patients, subdivided according to cytogenetic analysis. (E) median levels of IL-17A in CLL patients carrying the 11q22.3 deletion and/or the 17p13.1 deletion (12.24 pg/ml) and patients without these genetic aberrations (23.55 pg/ml; p = 0.016) (F) Th17 cell percentages in patients with del(11q22.3) or/and del(17p13.1) and patients without these unfavorable genetic aberrations (8.55% vs. 9.65%).
Figure 5.
Plasma IL-17A levels and percentage of Th17 cells in treatment patients.
(A) Plasma levels of IL-17A in patients who achieved hematological remission (CR: 19.73 pg/ml and PR: 17.47 pg/ml) after fludarabine-containing protocols in comparison to non-responding patients (PD: 9.09 pg/ml). (B) percentages of Th17 cells in the patients with progressive disease (3.04%) and those with complete (4.08%) or partial remission (3.83%). CR – complete remission; PR - partial remission; PD – progressive disease.