Assessment of peripheral blood and bone marrow cells apoptosis caused by purine analogues in patients with chronic lymphocytic leukemia in correlation with parameters of disease progression.

Chronic lymphocytic leukemia (CLL) is a heterogeneous disease with variable clinical course and prognosis. Therefore, the role of prognostic factors is very important, especially for identifying the group of patients who require intensive treatment. The aim of this study was to assess whether the rate of apoptosis caused by purine analogues differs between patients with better or worse prognostic factors. The experiments were preformed in cultures of blood and bone marrow obtained from CLL patients. The cultures were supplemented with cladribine and fludarabine. We determined the percentage of caspase-3-positive cells and the BCL-2/BAX ratio, and subsequently these apoptosis markers were correlated with the expression of ZAP-70 and CD38, lymphocyte counts, lactate dehydrogenase and beta(2)-microglobulin levels and clinical stage according to the Rai classification. The results showed that bone marrow cells are more sensitive to apoptosis caused by purine analogues than cells derived from blood, supporting the idea that these two compartments have different proliferative statuses. The cells from ZAP-70-positive patients seem to enter apoptosis more readily than those from ZAP-70-negative patients; thus, ZAP-70-positive patients are more likely to benefit from treatment with purine analogues.

High dose methotrexate population pharmacokinetics and Bayesian estimation in patients with lymphoid malignancy.

The purpose of present study was to develop a population pharmacokinetic model of high dose methotrexate (HD-MTX) infusion in patients with lymphoid malignancy, to investigate the biological and clinical covariates related to the drug distribution and elimination. It is also the purpose to propose a limited sampling strategy (LSS) for the estimation of the time above the threshold (0.2 micromol.L(-1)). A total 82 patients with lymphoid malignancy were involved in the study. A pharmacokinetic model was developed using nonlinear mixed-effect model. The influence of demographic characteristics, biological factors, and concurrent administration were investigated. The final predictive performance was validated by bootstrap and cross-validation. Bayesian estimation was evaluated. The pharmacokinetics of HD-MTX was described by a two-compartment model. The pharmacokinetic parameters and the inter-individual variability were as follows: the clearance CL, 7.45 L.h(-1) (inter-individual variability 50.6%), the volume of the central and peripheral compartment V(1), 25.9 L (22.5%), V(2), 9.23 L (97.8%), respectively, and the intercompartmental clearance Q, 0.333 L.h(-1) (70.4%). The influence of serum creatinine on CL and weight on V(1) was retained in the final model. The protocol involved one sampling time at 44 h after the start of the infusion, allowing one to predict the time at which the MTX concentration reached the expected threshold (0.2 micromol.L(-1)). Serum creatinine and weight showed significant influence on methotrexate CL and V(1), respectively. Furthermore, a Bayesian estimation based on the covariates and 44 h sample was developed, allowing prediction of the individual methotrexate pharmacokinetic parameters and the time to 0.2 micromol.L(-1).

Thymic humoral factor in the assessment of T lymphocytics in a patient with T cell chronic lymphocyte leukemia.

Peripheral blood lymphocytes from a patient with T cell chronic lymphocytic leukemia were examined by a combination of cell markers (E and M rosettes, and surface Ig), the graft-vs.-host reaction, thymic humoral factor (THF) and scanning electron microscopy. It was not possible, at first, to determine by conventional methods whether the leukemic cells were of the B or T type, but the THF and graft-vs.-host reaction showed that T cell precursors were present. These cells were incompetent immature T cells that underwent maturation following treatment with THF. Three months later, the T cell nature of the disease was clearly demonstrated by the E rosette technique, although at this time the cells were no longer influenced by THF.

[Assessment of the quantity of T-lymphocytes and other rosette-forming cells in the blood of patients and healthy subjects]. / Osobennosti otsenki kolichestva T-limfotsitov i drugigh rozetkoobrazuiushchikh kletok v krovi zdorvykh i bol'nykh liudei

In using the reaction of spontaneous rosette formation to assess the immunity T-system one should not limit one-self to determination of the percentage content of the rosette-forming cells alone. Immune status can be properly judged by determination of the content of immunocompetent cells per one cubic mm of the blood of the given donor. When the reaction is staged on the leukocyte suspension not only lymphocytes, but also other cell forms with the capacity of binding sheep erythrocytes can be determined.