[The complex morphologic diagnostic of marginal zone lymphoma.]

The purpose of study: To determine possibilities of complex morphological diagnostic (routine cytology and histology, immunocytochemistry and immunohistochemistry, molecular genetics) of lymphoma of marginal zone. The study included 10 patients with diagnosis of lymphoma of marginal zone cells, established on the basis of application of morphological (cytological and histological), immunomorphologic (immunocytochemical and immunohistochemical) and molecular genetic techniques. The immunofenotyping was implemented using immunocytochemical technique (EnVision FLEX) applying monoclonal antibodies by DAKO manufacturer. The immunofenotyping was implemented by flow cytofluorometry technique (flow cytofluorometer FACS Calibur by Becton Dikinson, USA) using antibodies by DACO manufacturer labeled by fluorescent marks (FITC or RPE). The antibody panel included: common leukocytic antigen, common cytokeratins CD19, CD20, CD79a,CD10, Bcl2, Bcl6, CD23, CD34, TdT, CD3, CD4, CD5, CD8, CyclinD1, Ki67, κ, λ. The FISH technique was applied using probes Bcl2 FISH DNA Probe, Split Signal и MALT1 FISH DNA Probe Split Signal by DAKO manufacturer. In one patient a gene Bcl2 change was detected with purpose of differentiating diagnostic with follicular lymphoma. In three patients, diagnosing lymphoma of marginal zone was implemented by FISH technique using detection of translocation (11;18)(q21;q21), involving gene of inhibitor of apoptosis API2 and gene MALT1. The routine cytological analysis of ten cases permitted to establish an exact diagnosis of lymphoma only in five cases and with no indication that it is lymphoma of marginal zone. In two cases under routine cytological analysis only a suspicion about lymphoma was suggested. In all ten patients a positive expression of pan-B-cellular markers CD19, CD20, CD79a was observed both under immunocytochemistry and immunohistochemistry and flow cytofluorometry. In 6 patients (60%) a positive expression of gene Bcl2 was observed both under immunocytochemistry and immunohistochemistry and flow cytofluorometry. The comparison of immunocytochemistry and immunohistochemistry and flow cytofluorometry established absence of expression of CD5, CD3, CD10, CD34, CD23, Bcl6, TdT, cyclin D1. Under immunocytochemistry and immunohistochemistry in ten cases proliferative activity protein Ki-67made up no more than 30%. Under flow cytofluorometry clonality on light chains of immunoglobulins κ or λ (Igλ/Igκ) were established. Overall, correlation coefficient (r, p < 0,05) between immunocytochemistry and immunohistochemistry, flow cytofluorometry made up to 1. The four patients were applied FISH-reaction for adjustment of diagnosis. In patient with nodal lymphoma of marginal zone gene Bcl2 change was absent. In three patients with MALT-lymphoma a gene MALT1 change was established. Thereby, accuracy of routine cytological analysis at diagnosing lymphoma without indication of its type in case of lymphoma of marginal zone made up to 50%, sensitivity - 50%, specificity - 100%. The accuracy of immunofenotyping permitting diagnosing lymphoma of marginal zone made up to 100%, sensitivity - 100%, specificity - 100%. The correlation coefficient (r, p < 0,05) between data of immunocytochemistry and immunohistochemistry, flow cytofluorometry made up to 1. The accuracy, sensitivity and specificity of complex analysis (cytology, immunocytochemistry and FISH-technique) in diagnostic of lymphoma of marginal zone made up to 100%.

[Therapy for Burkitt's lymphoma according to the BL-M-04 protocol: 12-year experience].

AIM: To evaluate the efficiency and toxicity of the intensive Burkitt's lymphoma (BL) therapy protocol BL-M-04. SUBJECTS AND METHODS: A total of 70 patients diagnosed with BL, including 45 men and 25 women whose age was 15 to 62 years (median age 31 years), were followed up in 2003 to 2014. Stage I (according to S. Murphy) was diagnosed in 4 (5.7%) patients; II in 9 (12.9%), III in 25 (35.7%), IV in 11 (15.7%), and Burkitt's leukemia in 21 (30%). There were tumor involvements of the bone marrow and central nervous system in 23 (32.9%) and 15 (21.4%) patients, respectively. B symptoms were detected in 56 (80%) patients; enhanced lactate dehydrogenase (LDH) activity was found in 50 (78.1%) out of 64 patients; moreover, in 34 (56.2%) out of 64 patients, LDH activity was more than twice as high as the reference values. The median LDH activity was 2398 (238-20,300) U/I. Acute renal failure at disease onset was identified in 17 (24.2%) patients; chemotherapy was initiated in 8 patients during renal replacement therapy. The treatment was performed using the BL-M-04±R protocol (4 successive blocks of A-C-A-C±R). Six blocks of A-C-A-C-A-C with rituximab has been carried out in patients with bone marrow involvement since 2011. RESULTS: Sixty-two (89%) patients achieved complete remission. At this time, 6 patients died from therapy complications during remission induction; 2 patients were observed to have disease progression; 3 developed disease recurrence (2 patients had early recurrence; 1 patient developed recurrence 2 years after treatment). Five-year overall survival (OS) was 85%; 5-year relapse-free survival (RFS) was 95%. The Cox multivariate regression analysis revealed that Burkitt's leukemia and bone marrow involvement were independent factors that influenced OS and RFS. The poor somatic status (3-4 ECOC scores versus 0-2 scores) proved to be statistically significant for OS rather than RFS. CONCLUSION: Despite the optimistic results obtained by our study group, there is a need to further improve BL treatment protocols and to elaborate novel approaches to therapy particularly for older patients and patients with Burkitt's leukemia.