Assessment of the pathway of apoptosis involving PAR-4, DAXX and ZIPK proteins in CLL patients and its relationship with the principal prognostic factors.

Par-4 (prostate apoptosis response-4) protein was originally found upregulated in prostate tumor cells undergoing apoptosis. Then it was further identified as a proapoptotic protein upregulated both in normal and leukemic lymphocytes. The aim of our study was to assess PAR-4 protein expression in the B cells of CLL patients and to examine its relationship with the expression of other proteins involved in the apoptosis process, such as DAXX, ZIPK and BCL-2. We found a positive relationship between PAR-4 and BCL-2 protein expression. Additionally, there was a positive correlation between PAR-4 and both DAXX and ZIPK protein expression. The results of our research were also analyzed in association with the principal CLL prognostic factors. There was a positive correlation between the expression of PAR-4 protein and the lactate dehydrogenase (LDH) serum concentration (p < 0.005). The expression of PAR-4 protein in B cells correlated positively with the percentage of CD38(+) cells (p < 0.05), as well as with CD38(+)/ZAP-70(+) cells (p < 0.05). Moreover, we found a close relationship between LPL protein expression or LPL/ADAM29 MFI ratio and PAR-4 protein expression. Our results confirm the significance of apoptosis deregulation in CLL, and suggest a possible relationship between PAR-4 expression and the clinical course of the disease. This however requires further investigation.

Natural killer-like T CD3+/CD16+CD56+ cells in chronic lymphocytic leukemia: intracellular cytokine expression and relationship with clinical outcome.

The clinical significance of NK-like T CD3+/CD16+CD56+ cells in chronic lymphocytic leukemia (CLL) is still a subject of controversy. There are few previous descriptions that this cell population can be qualitatively or quantitatively deficient in CLL patients. In the present study we investigated the clinical value of CD3+/CD16+ CD56+ cells as predictors of disease progression. We assessed the frequencies of CD3+/CD16+CD56+ cells by the flow cytometry in a group of 300 CLL patients. The percentage of CD3+/CD16+CD56+ cell population expressed as the percentage of CD3+ lymphocyte compartment showed an inverse correlation with ZAP-70 and CD38. Additionally, the CD3+/CD16+CD56+ showed an inverse correlation with LPL/ADAM29 ratio. Likewise, the ability of these cells to cytokine expression correlated with ZAP-70 expression. A positive correlation between percentage CD3+/CD16+CD56+ cells and TFS was found. The decreased percentage of these cells was associated with higher death risk in CLL patients. Furthermore, the percentage of CD3+/CD16+CD56+ cells was significantly decreased in patients who showed progression of disease. This study suggests that assessment of CD3+/CD16+ CD56+ cells may be helpful in determining a worsening of clinical course. Monitoring of these cell numbers and function may provide useful information for determining disease activity. Especially, it could be intormative to look at these cells in patients with stage 0 CLL. For this patient group immunological control and dysfunction are probably important factors.

BAFF and APRIL expression in B-cell chronic lymphocytic leukemia: correlation with biological and clinical features.

B-cell activating factor (BAFF) and a proliferation-inducing ligand (APRIL) are involved in normal B cell survival and differentiation. We analyzed BAFF and APRIL plasma levels in patients with B-cell chronic lymphocytic leukemia (B-CLL). We also tested intracellular BAFF and APRIL expression in B-CLL and evaluated their prognostic relevance. The percentage of leukemic B cells with intracellular APRIL or BAFF expression correlated significantly with adverse prognostic factors such as ZAP-70 and CD38. Moreover, we found a close relationship between LPL protein expression or LPL/ADAM29 MFI ratio and proportion of B-CLL cells with intracellular BAFF or APRIL expression. Furthermore, patients with a low percentage of leukemic cells with intracellular BAFF or APRIL expression had a significantly longer overall survival than those with a high proportion of APRIL or BAFF positive leukemic cells.

ZAP-70 and CD38 expression are independent prognostic factors in patients with B-cell chronic lymphocytic leukaemia and combined analysis improves their predictive value.

Recently identified biological risk factors in B-cell chronic lymphocytic leukemia (B-CLL) include ZAP-70 and CD38 expression. The present study was conducted to clarify whether a combined analysis could improve predictive impact of these two parameters. We examined the expression of ZAP-70 and CD38 by flow cytometry method in 217 newly diagnosed, consecutive, unselected and well characterized B-CLL patients in relation to laboratory parameters and clinical outcome. We confirmed that both ZAP-70 as well as CD38 were independent of prognostic factors. There was a significant correlation between the percentage of leukemic cells positive for ZAP-70 and the percentage of CD38+CD19+ cells (R=0.629; p=0.000001). Combined analysis of ZAP-70 and CD38 showed concordant results in 158/217 patients (72.8%), while in 59 patients the results were discordant (27.2%). A mean treatment free survival (TFS) was the longest in ZAP-70-CD38-patients (45.6 months, comparing to 13.6 months in ZAP-70+CD38+ group). Also a mean overall survival was the longest in ZAP-70-CD38- patients (224.7 months compared to 77.9 months in ZAP-70+CD38+ patients).

Apoptosis in B-CLL: the relationship between higher ex vivo spontaneous apoptosis before treatment in III-IV Rai stage patients and poor outcome.

B-cell chronic lymphocytic leukaemia (B-CLL) has been described as the progressive accumulation of mature-appearing B cells in peripheral blood and bone marrow, resulting from failed apoptosis rather than from alterations in cell cycle regulation. Recent investigations suggest that high WBC and lymphocyte counts result not only from defects in apoptosis, but also from cell proliferation. In this study, we aimed to examine the process of apoptosis in B-CLL patients before and during anti-cancer therapy, to answer the question of whether this parameter would presage the response to treatment and the clinical course of the disease. We found that ex vivo spontaneous apoptosis was higher in advanced-stage (III-IV acc. Rai) than in early-stage (I-II acc. Rai) patients. In I-II Rai stage patients the percentage of ex vivo apoptotic cells after chemotherapy was higher than that of apoptotic cells prior to treatment, whereas in III-IV Rai stage patients the percentage of ex vivo apoptotic cells after chemotherapy was lower than that of apoptotic cells before the anti-cancer therapy. The results of our study, in the context of the cited literature, suggest a relationship between higher ex vivo spontaneous apoptosis before treatment in advanced-stage patients with a higher proliferation of leukaemic cells and poor outcome.