[Hepatic involvement in patients with non-Hodgkins lymphoma]. / Postizení jater u nemocných s ne-Hodgkinovým lymfomem.

Non-Hodgkins lymphoma (NHL) represent the most frequent hematological malignancy with frequent extranodal involvement. We have identified 79 pts (4.6%) out of 1,712 patients with NHL, who were diagnosed in our center between 1999-2010. Five cases were primary extranodal lymphomas and we have observed one primary hepatic lymphoma (0.015%). The most frequent (61.3%) NHL subtype in our cohort was diffuse large B-cell lymphoma. B-NHL formed 92.4% of all lymphomas. We have observed high number of HBsAg positive patients (10%). The whole group have poor prognostic features with high number of patients (85%) with intermediate-high and high risk according to international prognostic index. The patients were treated with chemotherapy in 95%, B-NHL patients recieved immunochemotherapy with rituximab in 77%. The median progression free survival, resp. overall survival 4.6, resp. 8.4 years in the whole group and 1.4, resp. 8.4 years in diffuse large B-cell lymphoma were observed with median follow-up 4.5 years. The outcome of T-NHL patients was significantly worse with overall survival median 1.2 vs 8.4 years (p < 0.033). The patients with B-NHL treated by immunochemoterapy with rituximab had significant death risk reduction (HR 0.44, p = 0.03) compared to the patients treated with chemotherapy.

Dendritic cells and T lymphocyte interactions in patients with lymphoid malignancies.

Dendritic cell (DC) vaccination is an attractive approach to the treatment of patients with lymphoid tumors. To evaluate its feasibility, we have tested the functional properties of DC and T-lymphocytes in patients with treated and untreated chronic lymphocytic leukemia (CLL) and follicular lymphoma (FL). Healthy volunteers were used both as controls and as a source of cells for allogeneic mixed leukocyte reaction (MLR). In these reactions, dendritic cells from both untreated and treated patients were comparable to dendritic cells from healthy volunteers. In all the untreated patients studied, autologous dendritic cells promoted the survival and proliferation of both CD4 and CD8 lymphocytes (though the proliferation response was much better in the CD4 subset), whereas only 3 out of 5 treated patients were able to mount this response with CD4 lymphocytes and 4 out of 5 with CD8 lymphocytes. In 3 out of 5 untreated patients, pulsing of DCs with tetanus toxoid promoted a better CD4 response than was achieved with unpulsed DCs, while none of 5 treated patients had an additional response after pulsing with tetanus toxoid. None of patients studied, either treated or untreated, had a better CD8 response to pulsed DCs than to unpulsed ones. During CD4 lymphocyte proliferation, more CD4(+)CD25(hi) lymphocytes were generated in both treated and untreated patients than in healthy controls. Poor proliferation of cytotoxic cells and preferential proliferation of CD4(+)CD25(hi) T-regulatory cells in response to self and/or foreign antigens might be one of the mechanisms responsible for immunosuppression and impaired tumor surveillance in patients with lymphoid malignancies.

[The outcome of whole-body FDG-PET examination predicts the future of patients with diffuse large-cell lymphoma in the use of both intermediary staging and at the end of standard chemotherapy]. / Výsledek celotelového FDG-PET vysetrení predikuje osud nemocných s difuzním velkobunecným lymfomem jak pri pouzití v intermediárním stagingu, tak pri pouzití na konci standardní chemoterapie.

AIM: Response to the therapy is one of the most valuable prognostic factors. The response evaluation is performed by computer tomography as a standard tool. The introduction of FDG-PET whole body imaging allows to discriminate viable tumor and fibrotic changes in structural abnormalities. METHODS: We have performed retrospective analysis of 96 patients with diagnosis of diffuse large B-cell lymphoma (1999-2004) who were treated by anthracyclin based chemotherapy and FDG-PET was performed as a part of intermediate restaging (after 2nd-4th cycle, 69 patients) or/and at the end of standard chemotherapy (68 patients). RESULTS: The progression free survival (PFS) and overall survival (OS) at 3 years were the endpoints. Median follow up was 30 months. The PFS and OS resp. for PET negative pts at intermediate restaging was 80.7% and 97.6% compared to the 50.5% and 71.5 % resp. for PET positive patients. The relapse risk and death risk for PET positive patients was 4.8 and 6.4 resp. The PFS and OS resp. for PET negative pts at the end of chemotherapy was 81.7% and 94.7% resp. compared to the 29.4% (p < 0.0001) and 57.5% (p < 0.0001) resp. for PET positive patients. The relapse risk and death risk for PET positive patients was 7.0 and 12.9 resp. Predictive value of PET at intermediate as well at the end restaging was observed in IPI low group as well IPI high risk subgroups for both PFS and OS, except OS in high risk subgroup at intermediate restaging. CONCLUSION: The current analysis confirms predictive PET value for patients with DLBCL at intermediate as well at the end restaging. The question if and how to use the PET findings for tailoreing of therapy remains to be answered in prospective trials.

[Clinical importance of semi-quantitative monitoring of lymphomas using the comparative polymerase chain reaction]. / Klinická relevance semikvantitativního monitorování lymfomu uzitím komparativní polymerázové retezové reakce.

BACKGROUND: PCR techniques detecting interchromosomal translocation and clonal immunoglobulin gene rearrangement (IgH) as disease markers in non-Hodgkin's lymphomas (NHL) has been utilised past ten years. However, qualitative PCR detection of persisted minimal residual disease cannot provide clinically useful prognostic information and presently, quantitative approaches are required to predict patient outcome and assess response to the treatment. In some cases, "end-point" quantifying techniques, such as comparative PCR, are applicable and the relative estimation of differences in target quantity may serve in disease monitoring rather than absolute number of target copies. METHODS AND RESULTS: Our method of comparative PCR employs co-amplification of sequences of interest (clonal CDR3, bcl2/Jh) and the segment of Hras 1 gene(ras) as an internal standard. Serial dilutions of stored diagnostic DNAs from blood and bone marrow are examined in the same PCR and, after gel densitometry, the amount of initial target is assessed by comparing exponential products of co-amplification. The comparative PCR assay was utilized in monitoring of NHL patients cured either with conventional therapy, or with high-dose regimens and transplantation with stem cells, or with chimaeric anti-CD20 monoclonal antibody (Rituximab). Results from 50 monitored intervals obtained during several months up to several years were supplemented with clinical statements retrospectively. Some of patients became PCR-negative, reappearance of PCR-positivity was observed as well. The decrease or increase of disease marker corresponded to clinical observations. Results obtained from bone marrow were in agreement with those obtained from blood. CONCLUSIONS: End-point quantifying PCR comparative assay may provide an information on the increased risk of relapse and impact of the therapy. The predictive value of these methods depends on the frequency of sample taking and on the sensitivity of the method, which should be monitored in negative cases.

[Recombinant erythropoietin (r-HuEPO) in the treatment of anemia in multiple myeloma]. / Rekombinantní erytropoetin (r-HuEPO) v lécbe anémie u mnohocetného myelomu.

BACKGROUND: Anemia is common complication in multiple myeloma (MM). Its etiology is multifactorial-bone marrow infiltration, cytokines production, renal failure and effect of chemotherapy are main contributing factors. The red cell substitution therapy, which is administrated in 34 - 53% of patients, is frequently associated with the risk of well-known side effects. The use of recombinant erythropoietin (r-HuEPO) is a novel therapy in patients with MM. METHODS AND RESULTS: We have investigated the effect of r-HuEPO (Eprex. Cilag) in the group of 8 patients with MM. The growth factor was administrated in the dose of 150 U/kg. 3 times/ week s.c. The criterium of response was the increase of Hb levels of 20 g/L. All patients responded to r-HuEPO treatment. The medium period of response was 6,5 weeks. In two patients the doses of r-HuEPO could be reduced due to excellent effect of therapy. The energy level, ability to daily activities and overall quality of life significantly improved during the course of therapy. Neither effect of growth factor on thrombopoiesis and/or leukopoiesis nor serious adverse events due to r-HuEPO therapy were observed. The activity of underlying disease did not seem to be affected by r-HuEPO. In one patient the disease rapidly progressed after the end of study. The progression had some features of plasmablastic leukemia and leads to the death of patient. CONCLUSIONS: According to our experience, r-HuEPO is highly effective in the treatment of anemia in MM. The stimulation of erythropoiesis was associated with significant improvement of quality of life of patients. The effect of r-HuEPO treatment on activity of MM was not found, however, in one patient rapid progression of disease was observed after the end of study.

[The second reading of hematologic biopsies]. / Druhé ctení hematologických biopsií.

Occurrence of differences between the first and the second reading was compared in three time periods. The first from the years 1975-1989 showed disagreement in 34%, the second from 1990-1991 in 86%, the latest from 1994-1995 in 45%. The failures of the same sort repeated in 19 from 42 cases. They could be avoided by confining capacities of one's own department and using advantage of consulting reading by other pathologist. The second reading asked by an oncologist represents an independent urgent part of individual patient's treatment and should be generally supported and freed from any hindrance.

[Prognostic factors in non-Hodgkin's malignant lymphoma]. / Prognostické faktory u nehodgkinských maligních lymfomu.

A group of 271 patients with non-Hodgkin's malignant lymphomas from years 1975-1989 was tested using multivariant statistical analysis, from the point of view of the prognostic value their basic clinical and laboratory data. In the group of low grade malignancy lymphomas, the following factors showed a prognostic validity: clinical stage, general status, centrocytic histological subtype, and anaemia. In the group of high grade malignant lymphomas following one were set as prognostically important: general status, clinical stage and age. In particular histological subtypes of low grade malignancy statistically valid difference of survival was not proved, the centrocytic type excluded. However, the difference was found between these types and particular subtypes of high malignancy grade. Based on these results, the Cox's risk model was made, enabling us to define 4 risk groups according to the histological subtype and clinical stage: group A (risk less than 0.5), group B (risk = 0.5-1.0), group C (risk = 1.0-2.0), group D (risk greater than 2.0).

[Holter monitoring of patients treated with cytostatic agents]. / Holterovské monitorování nemocných lécených cytostatiky.

After cytostatic treatment severe arrhythmias, the development of angina pectoris and even the development of acute myocardial infarction and sudden death were observed. Therefore we made in 42 patients with malignant haematological disease treated with cytostatics 96 Holter monitorings of the electrocardiographic signal. The monitoring was made during the administration of cytostatics as well as during the time interval between the administration of combinations of cytostatics. In both instances (during the administration and during the interval between administration) we recorded a surprisingly high, mean all-day as well as maximal, heart rate. In the group monitored during administration of chemotherapeutic drugs we observed 5-8 hours after administration of cytostatics serious ventricular arrhythmias [incl. ventricular tachycardia], denivelization of the ST segment, paroxysms of supraventricular tachycardia. In the group monitored during the interval between administration of cytostatics the sick-sinus syndrome was recorded, as well as a passive nodal rhythm, disorders of the intraventricular conduction. The described changes are explained by the release of vasoactive substances after administration of cytostatics, by a change of the transmembrane calcium transport leading to an increased excitability of the heart muscle and possibly to coronary spasms and direct irreversible damage of the conduction system.

Labelling of a T lymphocyte subpopulation with monoclonal antibody bound to colloidal gold. Correlation with the indirect immunofluorescence method.

A method of direct labelling of human lymphocytes CD8 with a monoclonal antibody bound to colloidal gold particles is described. Results of labelling of isolated lymphocytes and of cells of leucocyte concentrate from haemolysed blood correlated well with those obtained by indirect immunofluorescence. Owing to its simplicity and rapidity, the described method is particularly suitable for clinical diagnostics.

Studies on nucleoli in rosetting T and B lymphocytes of the human peripheral blood.

The incidence of various nucleolar types was studied in human rosetting lymphocytes to provide an information on nucleolar types present in T and B lymphocytes of the peripheral blood. The results clearly demonstrate that both T and B lymphocytes of the peripheral blood mostly contain ring shaped nucleoli ("resting nucleoli") and less frequently other nucleolar types such as nucleoli with nucleolonemata or compact nucleoli ("active nucleoli") and micronucleoli ("inactive nucleoli"). Since all known nucleolar types and particularly micronucleoli may be observed in both T and B lymphocytes, nucleoli in these cells cannot indicate the type or origin of these cells but simply the state of the nucleolar RNA synthesis.

Staining pattern of acid non-specific esterase in lymphocytes, plasmocytes, monocytes, and leukemic cells.

The pattern of staining for ANAE in lymphocytes, plasmocytes, monocytes and leukemic cells has been studied and the effect of NaF and E 600 on this staining reaction has been investigated. In lymphocytes the transition occasionally occurring between a dot-like (T type) and granular positivity may cause difficulties when using this reaction as a marker of resting T lymphocytes. In plasmocytes as well as in myeloma cells a number of coarse granules was present in most cells and in some cases even large blocks of the reaction product occurred. NaF and E 600 inhibits already in lower concentration (2 mM and 10 micronM respectively) the strong diffuse reaction in monocytes, the effect on other cells being negligible. Using higher concentrations the inhibitory effect of NaF (20 mM) was more pronounced in plasmocytes in comparison with T lymphocytes which on the other hand, were, in general more sensitive to E 600 (10 mM). Partially diffuse or dense granular pattern of ANAE was found in a number of hairy cells. "Lymphoblastic" leukemias in children most frequently displayed negative or fine granular reaction in a varying number of blasts and similar findings were noted in the majority of cytochemically undifferentiated leukemias in adults. In some of them, however, a similar pattern of ANAE as in myeloblastic leukemias with dispersed or partial diffuse positivity was observed. The significance of these findings is briefly discussed.

E rosettes in lymphoproliferative diseases.

Results are given of investigations of E rosettes in patients with chronic lymphadenosis and malignant lymphoma. The quantity of E rosettes was expressed both as per cents and in absolute numbers calculated from the total amount of lymphocytes in peripheral blood. In the normal controls the percentage of E rosettes was 58% on average while in patients with chronic lymphadenosis it was found to have decreased significantly down to a mean of 6%. However, in absolute values the number of rosette forming lymphocytes was normal and even higher. In the group with malignant lymphoma the percentage decrease was less striking than in those with chronic lymphadenosis, however, the absolute number of rosette forming lymphocytes was always found to be lowered because of evident to considerable lymphopenia. A dynamic study of the observed changes might contribute to a knowledge of the pathophysiology of lymphoproliferative diseases.