OBJECTIVES: The impact of modern prognostic markers on clinical course of chronic lymphocytic leukaemia (CLL) in everyday practice has been not yet well defined, especially in large series of patients. Therefore, the goal of this study was to assess the influence of conventional as well as modern prognostic factors on overall survival (OS) and time to therapy (TTT) of patients with CLL. METHODS: We retrospectively analysed data of all patients consecutively entered into the databases of five large academic centres in the Czech Republic. The total of 1300 patients was included in the analysis. RESULTS AND CONCLUSION: Through the use of uniparametric analysis, it was determined that gender, clinical stage Rai II-IV, unmutated IgVH status, deletion 17p (for both 5% and 20% cut-off), deletion 11q, ZAP-70 positivity and high expression of CD38 had significant negative influence on OS. TTT was significantly influenced by gender, Rai stage, IgVH status, deletion 11q, deletion 17p, deletion 13q and CD38 expression. Multiparametric analysis revealed that OS was significantly influenced by gender, age, IgVH status and deletion 17p. If only patients who died of CLL were included, gender, age, Rai stage, IgVH status and deletion 17p had significant influence on OS. Based on our results, the examination of biological prognostic markers can give an insight into the possible disease evolution in daily clinical practice. Biological prognostic markers are, however, not ready (maybe except deletion 17p in younger patients) to be used for guidance of therapy at least outside of clinical trials.

• MeSH
• analýza přežití MeSH
• antigeny CD38 krev   MeSH
• chromozomální delece MeSH
• chronická lymfatická leukemie etiologie   genetika   imunologie   patologie   MeSH
• difúzní velkobuněčný B-lymfom etiologie   MeSH
• dospělí MeSH
• geny pro těžké řetězce imunoglobulinů MeSH
• lidé středního věku MeSH
• lidé MeSH
• lidské chromozomy, pár 11 genetika   MeSH
• lidské chromozomy, pár 17 genetika   MeSH
• membránové glykoproteiny krev   MeSH
• mutace MeSH
• nádorové biomarkery krev   MeSH
• prognóza MeSH
• progrese nemoci MeSH
• protein-tyrosinkináza ZAP-70 krev   MeSH
• retrospektivní studie MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• staging nádorů MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

INTRODUCTION: It is widely accepted that expression of ZAP-70 in chronic lymphocytic leukemia (CLL) remains stable in time. However, data supporting this notion are surprisingly scarce. Therefore, we assessed expression of ZAP-70 in serial samples taken during the course of the disease. PATIENTS AND METHODS: We studied 44 patients with CLL diagnosed according to NCI-WG criteria (34 men, 10 women, median age 62, range, 36-81). A total of 104 samples were examined; all patients had at least two measurements. Median interval between the first and the second sample was 13 months (range, 2-36). ZAP-70 expression was detected by flow cytometry using phycoerythrin-conjugated monoclonal antibody clone 1E7.2 and negative isotype control. Twenty percent of positive cells were considered as the threshold of positivity. RESULTS: Significant change in ZAP-70 expression (i.e. from positivity to negativity and vice versa) was detected in 15/44 patients (34%). Interestingly, 7/8 patients whose ZAP-70 expression converted to positivity had unmutated IgVH genes. In addition, the conversion was accompanied by clinical progression or relapse in all but one patient. On the other hand, 5/7 patients with loss of ZAP-70 had stable clinical course. One patient became ZAP-70-negative during treatment with prednisone for autoimmune hemolytic anemia. CONCLUSIONS: In contrast to commonly accepted opinion, significant change in ZAP-70 expression in time was detected in a substantial proportion of our patients with CLL. While the conversion to ZAP-70 negativity was found predominantly in patients with stable disease, change to positivity was typical in patients with unmutated IgVH genes at the time of progression or relapse. Based on our pilot results, repeated assessment of ZAP-70 expression might be especially useful at the time of progression or relapse in patients who were initially ZAP-70-negative.

• MeSH
• chronická lymfatická leukemie enzymologie   etiologie   MeSH
• dospělí MeSH
• geny pro těžké řetězce imunoglobulinů MeSH
• lidé středního věku MeSH
• lidé MeSH
• mutace MeSH
• nádorové biomarkery krev   MeSH
• pilotní projekty MeSH
• progrese nemoci MeSH
• protein-tyrosinkináza ZAP-70 krev   MeSH
• recidiva MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Chronická B-lymfocytová leukémia (B-CLL) je jedna z najčastejších chorôb lymfatického systému u dospelých pacientov. Je známa rôznorodým klinickým priebehom – od benígnych až po infaustné formy. Pre spresnenie diagnózy a určenie správnej terapie sú potrebné spoľahlivé prognostické markery. Perspektívnym sa ukázal byť proteín ZAP-70, ktorý môže byť vo zvýšenej miere exprimovaný leukemickými bunkami pacientov s B-CLL. Dáva sa do súvisu s horšou prognózou pacienta vyžadujúcou intenzívnejšiu terapiu už v začiatočných štádiách. Základ prezentovanej štúdie tvorilo 20 prípadov biopsií kostnej drene od pacientov s diagnostikovanou B-CLL a 20 kontrolných prípadov bez nádorovej transformácie. Pri spracovaní materiálu bol okrem štandardnej imunohistochemickej metódy využitý aj amplifikačný systém (firmy DakoCytomation©). Výsledok bol hodnotený semikvantitatívne; ako negatívny (menej ako 5 % pozitívnych leukemických buniek), pozitívny (viac ako 30 % pozitívnych leukemických buniek) a nepravidelne pozitívny (medzi 5 % a 30 % pozitívnych leukemických buniek) a následne korelovaný s klinickým hodnotením prognózy pacienta. Výsledok pozitivity bol kontrolovaný aj morfometrickou analýzou, určením plochy pozitivity markera ZAP-70 voči celkovej ploche jadier v preparáte. V kontrolných prípadoch bola pozorovaná pozitivita markera ZAP-70 viazaná čisto na T-lymfocytov. Miera jadrovej pozitivity proteínu ZAP-70 pozorovaná v transformovaných B-lymfocytoch vykazovala signifikantnú súvislosť s horšou klinickou prognózou pacienta v prípadoch ak bola hodnotená ako jednoznačne pozitívna, resp. negatívna pri oboch spôsoboch spracovania preparátu. Prípady s nepravidelnou pozitivitou nevykazovali jednoznačnú klinickú koreláciu. Použitie amplifikačného systému s možnosťou využiť nižšiu koncentráciu primárnej protilátky a potlačiť pozitivitu pozadia, zvýšilo kontrast nálezu a redukovalo počet prípadov s hodnotenou nepravidelnou pozitivitou z 30 % (pri bežnom imunohistochemickom spracovaní) na 10?%. Možno konštatovať, že ZAP-70 predstavuje hodnotný prognostický marker chronickej B-lymfocytovej leukémie. Jeho hodnotenie pomocou imunohistochémie je vhodnou metódou pre klinickú prax. Problematickým sa však stáva hodnotenie hranične a nepravidelne pozitívnych prípadov, pri ktorých môže byť nápomocným využitie histochemického amplifikačného systému. Minimalizácia výskytu falošnej pozitivity, resp. negativity vyžaduje striktné kritériá pri hodnotení nálezu.

Chronic B-lymphocytic leukemia (B-CLL) is one of the most frequent diseases of the lymphatic system in adults. It is known for its variable course – from benign to prospectless forms. Reliable prognostic markers are needed to precify the diagnosis and the correct therapy. The ZAP-70 protein, as a prospective marker, can be highly expressed in leukemic cells of B-CLL patients. It is related to a worse prognosis requiring intense therapy from the beginning. The present study includes 20 cases of bone marrow trephine biopsy from patients with diagnosed B-CLL and 20 control cases without neoplastic infiltration. The specimens were investigated with standard immunohistochemical technique and with the use of the amplification system (DakoCytomation©). The results were evaluated semiquantitatively as negative (less than 5 %), positive (above 30 %), and irregularly positive (5–30 % of positive leukemic cells) and consecutively correlated with clinical evaluation of prognosis of the patient. The evaluation of the positivity was controlled also by morphometric analysis by determination of the area of ZAP-70 positivity related to the whole area of nuclei in the section. In the control specimens the ZAP-70 positivity was restricted to T-lymphocytes only. The level of the nuclear positivity of ZAP-70 protein detected in transformed B-lymphocytes showed significant correlation with the worse or the better clinical prognosis, respectively. Cases with irregular positivity did not show unambiguous clinical correlation. The use of the amplification system allowed to apply lower concentration of the primary antibody, reduction of background staining and increased the contrast of the findings leading to reduction of irregularly positive cases from 30 % (with routine histochemistry) to 10 %. It can be concluded that ZAP-70 represents a valuable prognostic marker for the chronic B-lymphocytic leukemia. Its evaluation by histochemistry is a suitable method for clinical praxis. Problematic may be the evaluation of borderline irregularly positive cases, in which the use of the histochemical amplification system is helpful. Strictly determined criteria are needed for limitation of inaccurate interpretation of the results.

• MeSH
• chronická lymfatická leukemie diagnóza   krev   MeSH
• diagnostické techniky molekulární metody   využití   MeSH
• financování organizované MeSH
• imunohistochemie metody   využití   MeSH
• lidé MeSH
• mikroskopie metody   využití   MeSH
• nádorové biomarkery izolace a purifikace   krev   MeSH
• nádorové proteiny izolace a purifikace   krev   MeSH
• prognóza MeSH
• protein-tyrosinkináza ZAP-70 izolace a purifikace   krev   MeSH
• Check Tag
• lidé MeSH

• MeSH
• antigeny CD38 genetika   izolace a purifikace   krev   MeSH
• chronická lymfatická leukemie diagnóza   etiologie   genetika   MeSH
• cytogenetické vyšetření metody   využití   MeSH
• genetické markery genetika   imunologie   MeSH
• klinické laboratorní techniky trendy   využití   MeSH
• lidé MeSH
• mutace genetika   imunologie   MeSH
• nádorové biomarkery izolace a purifikace   krev   MeSH
• prognóza MeSH
• protein-tyrosinkináza ZAP-70 genetika   izolace a purifikace   krev   MeSH
• regulace genové exprese u leukemie genetika   imunologie   MeSH
• těžké řetězce imunoglobulinů genetika   izolace a purifikace   krev   MeSH
• Check Tag
• lidé MeSH