AIM:To ascertain indications to standard (CHOP-21/R-CHOP-21) and intensive (mNHL-BFM-90) treatment in patients with diffuse large B-cell lymphosarcoma (DLBCL) with involvement of lymphoid organs.MATERIAL AND METHODS:The trial, performed from January 2002 to December 2010, enrolled 139 DLBCL patients with affected lymph nodes (LN), tonsils, spleen, bone marrow (BM). The diagnosis was made according to WHO criteria. The patients were examined according to the protocol of lymphoproliferative diseases. Biopsy material from all 139 patients was studied immunohistochemically on paraffin blocks (LN, tonsils, spleen, BM) using a wide panel of antibodies. The same examinations of BM were made in all 18 cases of BM involvement. Cytogenetic examination was performed in 106 patients: 48 standard cytogenetic tests, 139 - FISH for t (14;18) as well as rearrangement of locus 3q27. Patients with a poor prognosis (n = 86, 61.8%) received intensive therapy according to mNHL-BFM-90 program. The signs of a poor prognosis were the following: massive tumor (tumor size more than 7.5 cm), invasion into the adjacent organs or tissues, stage III-IV disease by Enn-Erbor, high concentration of LDG. Patients without a poor prognosis (n = 53, 38.2%) received standard treatment CHOP-21 (n = 28) or R-CHOP-21 (n = 25).RESULTS:A complete remission without recurrences was achieved in all 53 patients without signs of unfavourable prognosis (100%). Overall 5-year survival was 96%, 2 patients died in remission of other causes. Of 86 patients with a poor prognosis a complete remission was achieved in 64 (74.4%) patients. Overall and recurrence-free 5-year survival was 65 and 86%, respectively.CONCLUSION:Standard treatment provided long-term complete remission in all the patients without poor prognosis. Intensive (mNHL-BFM-90) treatment produced the best results in generalized lesion without BM involvement. Overall 5-year survival was 84% in these patients and 12% in patients with BM involvement.

Aim. To diagnose diffuse large B-cell lymphosarcoma (DLBCLS) with primary involvement of the mediastinal lymph nodes (LN) and to evaluate the efficiency of aggressive polychemotherapy (PCT).Subjects and methods. The study included 15 patients (6 men and 9 women aged 18 to 70 years; median 38 years) followed up at the Hematology Research Center, Russian Academy of Medical Sciences, in 2004 to 2009. Three and 12 patients had Stages II and IIE DLBCLS, respectively. B symptoms were found in 14 (93.4%) patients. Increased lactate dehydrogenase (LDH) concentrations were detectable in 14 (93.4%) patients; tumors of 10 cm or more (bulky disease) were seen in 11 (73.3%). Enlarged cervical, supraclavicular, and axillary lymph nodes were found in 9 (60%) patients; lung involvement via extension in 9 (60%), and invasion into the pericardium in 5 (33.3%) and soft tissues of the anterior thoracic wall in (13.3%). There were no signs of involvement of extranodal organs at the moment of diagnosis. All the 15 patients received PCT according to the modified NHL-BFM-90 program: 4 to 6 courses depending on the response to the therapy; 10 (66.6%) and 5 (33.3%) patients had 4 and 6 courses, respectively; for consolidating purpose, 11(78.5%) patients were prescribed radiotherapy applied to the mediastinum in a cumulative dose of 36 Gy due to the fact that they had a residual mass.Results. Thirteen (86.6%) patients achieved a complete remission (CR). Primary PCT resistance was confirmed in one case. Another patient was stated to have near-complete remission. No recurrences were notified during the follow-up. The mean CR duration was 24.5 (range 2-49) months.Conclusion. DLBCLS with primary LN involvement is an individual nosological entity to be differentiated from primary mediastinal large B-cell lymphosarcoma. In most cases, DLBCLS shows signs of a poor prognosis, which makes it necessary to perform aggressive PCT.

Immunocytochemical staining with specific antibodies was used to study the expression of three nucleolar proteins (fibrillarin, B23/nucleofozmin, and SURF6), which were involved in pRNA maturation, in the lymphoid cells of healthy individuals and patients with lymphoproliferative diseases and to compare it with the expression of the known proliferation marker Ki-67 protein. The results indicated that fibrillarin was detectable at the comparable level in the lymphoid cells of the patients and in the peripheral blood lymphocytes of the healthy individuals. In one fourth of the patients, the proportion of cells containing B23/nucleofozmin was noticeably higher than that in the lymphocytes of donors; however, there was no great difference in patients with different types of the disease. The number of SURF6-positive cells was directly correlated with that of Ki-67-positive cells. The maximum level (47-67%) of SUR6-positive lymphoid cells was found in splenic lymphosarcomas and mantle cell lymphoma. The findings suggest that SURG6 protein may be of additional diagnostic and prognostic value.

The aim of the study was to characterize the T/NK lymphocytic leukemias cases and identify their variants, to evaluate clinical features and response to therapy. The study was carried out in 35 patients (12 men and 23 women) aged 18-71 years. Five groups of patients were distinguished by the results of the study. The groups differed by immunophenotype and disease course, this fact confirming the heterogeneity of large granular lymphocyte (LGL) T/NK lymphocytic leukemias. The disease manifested without cytopenia. in groups 1-3. No cytostatic therapy was needed for a long time in patients with these variants who developed no manifest cytopenia (1/3 of this patient population). The periods of observation without therapy surpassed 70 months. Patients with manifest cytopenia (2/3 of patients) received immunosuppressive therapy (cytostatics; or cyclosporin A). Positive effect of cytostatics was attained only after long-term therapy. Splenectomy led to reduction of the cytopenia syndrome, but only 2 of 8 patients subjected to splenectomy needed no further cytostatic therapy. It seems that aggressive course of the disease is not frequent and is intrinsicic of the rare forms observed in the studied patient population. These are mainly double positive (CD4(+)CD8(+)) LGIL T/NK lymphocytic leukemias. Presumably, this categories of patients should receive the campas drug at the first stage of therapy, as cytostatics are ineffective.

Nosological forms and survival of patients with T/NK cellular lymphatic tumors. Vinogradova Yu. E., Lutsenko I. N., Kremenetskaya A. M., Samoilova R. S., Kaplanskaya I. B., Vorobyov I. A., Zingerman B. V., Nikitin E. A., Kravchenko S. K. Hematology Research Center, Moscow. The T and NK cell lymphomas/leukemias in patients observed during 5 years were verified. 151 patients (74 men and 77 women, age 15-89 years) were included in the study. 24.4% patients were with T-cellular lymphomas of the skin, 22.5% with nodal peripheral T-cellular lymphomas, 16.6% with early (lymphoblastic) T-cellular lymphosarcomas, 14.5% with extranodal T-cellular lymphomas, 14.5% with large granular lymphocyte (LGL) T-lymphocytic leukemia. The survival median for patients with T-cellular lymphomas of the skin was 108 months, for patients with extranodal T-cellular lymphomas 86 months, in peripheral T-cellular lymphoma it was 46 months, and in early (lymphoblastic) T-cellular lymphoma/leukemia 43 months. The survival median in the group of patients with LGL T-lymphocytic leukemia could not be estimated during the mean period of observation of 72 10.5 months. The proportion of nosological forms and prognosis of T/NK lymphatic tumors are similar obtained in the European countries and in the USA, except the few cases of angioimmunoblastic lymphoma and the absence of male patients predominance. The NK lymphocytic lymphomas/leukemias were rare.

AIM:To compare efficacy and toxicity of conservative therapy (different programs of polychemotherapy) of gastric lymphosarcoma conducted for the last 10 years in Hematological Research Center of the Russian Academy of Medical Sciences.MATERIAL AND METHODS:The study included 63 patients (40 females and 23 males aged 14 to 78 years, mean age 49 years) with primary diagnosis of gastric lymphosarcoma (GL). Of them, 56 (89%) patients had diffuse large B-cell lymphosarcoma (DLBCL) and 7 (11%) had gastric Berkitt's lymphoma (BL). Only detection of t(8;14) with rearrangement of c-myc gene provided accurate diagnosis of gastric BL. By the treatment DLBCL patients were divided into two groups: 44 patients of group 1 received polychemotherapy (PCT) according to CHOP scheme or in combination with radiotherapy and surgical treatment; 12 patients of group 2 were treated according to modified program mNHL-BFM-90, without surgical or radiation treatment. Of 7 patients with gastric BL 5 patients received treatment according to a modified program mNHL-BFM-90 and 2 patients were given CHOP because of DLBCL misdiagnosis without cytogenetic detection of t(8;14).RESULTS:Overall survival in group 1 was 73% in mean follow-up 61 months. The survival depended only on initial factors of poor prognosis (PPF): tumor size over 10 cm, Ann-Arbor stage higher than IE, B-symptoms, elevated level of LDH. Overall survival of 18 gastric DLBCL patients without PPF reached 94%, of 26 patients with PPF - 60%. Lethality due to side effects was 4% (2 patients), primary resistance was 14% (6 patients), recurrence arose in 9% (4 patients). Overall survival in group 2 was 100% in mean remission duration 18 months, was unrelated to PPF (10 of 12 patients) but correlated with high toxicity. 5 BL patients treated with a modified mNHL-BFM-90 program achieved remission (a mean follow-up at present is 1 to 50 months, mean 24 months). 2 BL patients treated with CHOP died for a year.CONCLUSION:Gastric lymphosarcomas are sensitive to chemotherapy, thereby PCT only is effective in most patients. PPF in gastric DLBCL were responsible for poor outcome in 40% patients in CHOP treatment. The modified program mNHL-BFM-90 can produce up to 100% complete long-term remissions in therapy of gastric lymphosarcoma in adults both in BL and DLBCL patients. A cytogenetic examination of c-myc gene rearrangement is obligatory before initiation of PCT of gastric lymphosarcoma.

AIM:To characterize clinical symptoms, course, immediate and long-term treatment results in young patients with hair cell leukemia (HCL).MATERIAL AND METHODS:The data on 41 HCL patients were analysed. The diagnosis was made by standard diagnostic protocol for HCL detection.RESULTS:The analysis of the age of 160 HCL patients studied demonstrated high (26%) incidence of HCL at young age. Young patients with HCL had special clinical manifestations and specific long-term outcomes of treatment with standard schemes.CONCLUSION:Differences in occurrence of recurrences after standard therapy make it necessary to consider young HCL patients as a separate group who need adjuvant treatment to prolong remission.

Aim. To compare efficacy of NHL-BFM-90 and CHOP-like courses in the treatment of anaplastic large cell lymphoma (ALCL). Material and methods. Twenty-two patients with ALCL participated in the study. The diagnosis was made basing on the findings of clinical, device, morphological, immunohistochemical and molecular-genetic examinations with application of a panel of monoclonal antibodies to CD30, ALK, CD3, CD4, CD8, CD7, CD34, CD15, CD68, CD20, CD45RO, CD45RA, Ki-67. 14 cases of 22 were negative by kinase of anaplastic lymphocytes (ALK-) and 8 were positive (ALK+). Mean age of ALK-ALCL patients was 39.6 + 4.1 years, of ALK+ALCL patients - 23.4 + 2.6 years. 14 patients were treated by the protocol NHL-BFM-90, 8 were initially treated with other schemes (CHOP, MACOP-В, BEACOPP and others). All 14 patients treated according to NHL-BFM-90 had ALCL stages III-IV with B-symptoms. 12 patients who completed treatment by the above protocol achieved complete remission after the forth course, 2 patients failed the treatment. Of 8 ALCL patients treated initially according to other schemes, a complete remission was achieved in 4 patients (2 had stage II). One of 4 patients with remission had recurrence. Four patients who had failed to achieve complete remission died of the disease progression. Conclusion. ALCL occurs more frequently in young and middle-aged patients. The disease has an aggressive course with rapid generalization. For such processes it is more preferable to use a modified protocol NHL-BFM-90.

Microarrays are one of the latest breakthroughs in experimental molecular biology, which allow monitoring of gene expression for tens of thousands of genes in parallel and are already producing huge amounts of valuable data. Analysis and handling of such data is becoming one of the major bottlenecks in the utilization of the technology. The raw microarray data are images, which have to be transformed into gene expression matrices – tables where rows represent genes, columns represent various samples such as tissues or experimental conditions, and numbers in each cell characterize the expression level of the particular gene in the particular sample. These matrices have to be analyzed further, if any knowledge about the underlying biological processes is to be extracted. In this paper we concentrate on discussing bioinformatics methods used for such analysis. We briefly discuss supervised and unsupervised data analysis and its applications, such as predicting gene function classes and cancer classification. Then we discuss how the gene expression matrix can be used to predict putative regulatory signals in the genome sequences. In conclusion we discuss some possible future directions.

AIM:To investigate characteristics of the course and efficacy of treatment of diffuse large B-cell lymphosarcoma (DLBL) with primary lesion of the spleen.MATERIAL AND METHODS:From 1998 to 2006, primary splenic lesion was registered in 15 of 120 patients with DLBL and affected lymph nodes (LN), spleen and Waldeyer's ring. The diagnosis was made according to WHO criteria. Of them 14 patients had splenectomy as the first stage of therapy. The operation was followed with 6 to 8 courses of CHOP-21 (8 patients), 4 courses of R-CHOP-21 and radiotherapy (one patient). One patient received 7 courses of CHOP-21 followed by splenectomy. Because of the presence of several signs of unfavourable prognosis 5 patients under 60 years were given intensive therapy: 4-6 courses of the modified program NHL-BFM-90, 2 of 5 patients received radiotherapy.RESULTS:All the patients with primary DLBL of the spleen had two and more signs of unfavourable prognosis: elevated concentration of serum LDG, size of the tumor more than 10 cm, high proliferative activity of tumor cells, B-symptoms, severe condition. Seven patients had centroblastic, 8--anaplastic variants of DLBL. Tumor cells in primary DLBL of the spleen had no specific immunophenotype. Complete remission of the disease was achieved in 9 (90%) of 10 patients treated on programs CHOP-21, R-CHOP-21, in 4 of 4 patients on the modified program NHL-BFM-90. Mean follow-up was 39.3 months (from 7 to 103 months).CONCLUSION:For primary DLBL of the spleen characteristic are long-term remissions on first line therapy according to CHOP-21 program irrespective of morphology and immunophenotype.

Introduction: According to the new WHO classification for Non-Hodgkin's lymphomas developed in 2001, the diagnosis of many lymphoma patients has to be reevaluated. The existing information on therapeutic options for patients with these rare malignancies prior to development of WHO classification has been unreliable. The T-cell malignancies are associated with poor outcome, and it is important to identify patients that could benefit from novel intensive therapies.

Immunophenotypical analysis of the lymph node lymphoid cells was carried out in 85 patients (35 with nontumorous. lymphadenopathies, 40 with various lymphoproliferative diseases, and 10 with cancer metastases in the lymph nodes). For differential diagnosis, the phenotypical characteristics of lymphoid cells in the lymph nodes with histologically verified follicular hyperplasia (20 cases) and follicular lymphoma (24 cases), and of paracortical hyperplasia (5 cases) and T-cell lymphomas (6 cases) were evaluated. In addition, immunophenotype of lymphocyte populations in the lymph nodes of patients with granulomatous lymphadenitis (10 cases), Hodgkin's disease (10 cases), and cancer metastases was evaluated; these are diseases when reactive lymphoid population constitutes the bulk of hyperplastic lymph nodes, while the presence of granulomas, Berezovsky-Sternberg cells or cancer cells usually detected in histological preparations is diagnostically significant. Specific immunophenotypes of follicular and paracortical reactive hyperplasia of the lymph nodes in comparison with B-cellular follicular and T-cellular lymphomas were detected, which permit the differential diagnosis between nontumorous and tumor lymphadenopathies in the majority of cases. Reactive populations in granulomatous lymphadenitis, Hodgkin's disease, and cancer metastases are characterized. Potentialities and limitations of immunophenotypical analysis of the lymphoid populations are evaluated.