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The properties of bone marrow (BM)-derived multipotent mesenchymal stromal cells (MSCs) are altered in the patients with the diffuse large B cell lymphoma (DLBCL) without BM involvement. It was suggested that plasma from the patients contains soluble factors that affect MSCs. Plasma and BM-derived MSCs from the DLBCL patients at the onset of the disease and one month after the end of treatment were studied. Concentration of the plasma cytokines and gene expression in the MSCs were evaluated by the Bio-Plex Pro Human Cytokine Panel kit to measure 27 analytes and real-time PCR. Plasma and MSCs from the healthy donors were used as controls. Analysis of cytokines in the plasma from healthy donors and patients before and one month after the end of treatment revealed significant differences in the concentration of 14 out of 27 cytokines. Correlations between the levels of secreted cytokines were altered in the plasma from patients indicating that the immune response regulation was disturbed. Cultivation of the MSCs from the healthy donors in the medium supplemented with the plasma from patients led to the changes in the MSC properties, similar to those observed in the MSCs from patients. The BM-derived MSCs were shown to participate in the humoral changes occurring in the DLBCL patients. For the first time, it was shown that the precursors of the stromal microenvironment - multipotent mesenchymal stromal cells - are altered in the patients with DLBCL without bone marrow involvement due to the humoral effect of the tumor and the response of organism to it. Comprehensive analysis of the results shows that, when remission is achieved in the patients with DLBCL, composition of the plasma cytokines normalizes, but does not reach the level observed in the healthy donors. The discovery of a new aspect of the effect of the tumor B-cells on the organism could help to reveal general regularities of the humoral effect of various tumors on the bone marrow stromal cells.

Aim. Pharmacoeconomic analysis of R-DA-EPOCH and R-mNHL-BFM-90 combination immunochemotherapy in patients with prognostically unfavorable diffuse large B-cell lymphoma within randomized multi-center clinical trial DLB-CL-2015. Materials & Methods. The pharmacoeconomic analysis conducted between September 2018 and February 2020 was based on the treatment data of 22 patients enrolled in the DLBCL-2015 randomized multi-center clinical trial. This paper deals with the estimation of treatment outcomes in only one center, i.e., the National Research Center for Hematology. The R-DA-EPOCH induction therapy was administered to 14 out of 22 patients, 8 patients received the R-mN-HL-BFM-90 block treatment. Within the R-DA-EPOCH group the second-line therapy was administered subsequently to 5 (36 %) out of 14 patients with partial remission or disease progression. The R-mNHL-BFM-90 treatment resulted in no need to assign second-line regimens. At the first stage, the efficacy of the compared induction therapy regimens was assessed. The next stage of the pharmacoeconomic study sought to analyze only the direct medical costs associated with the whole chemotherapy process. Further, the cost-effectiveness analysis was carried out, which allowed to estimate the financial resources necessary to achieve 1 case of complete remission (CR). A pharmacoeconomic decision-tree model was developed. Results. CR was achieved in all 8 patients (100 %) who received the R-mNHL-BFM-90 block treatment. In the R-DA-EPOCH group CR was achieved only in 9 (64 %) out of 14 patients. The total mean cost of achieving 1 CR case per patient at all stages of diagnosis and chemotherapy with account for bed turnover (induction, second-line therapy, total supportive care) using R-mNHL-BFM-90 was 1,640,757 rubles, whereas in the R-DA-EPOCH group it was 1,469,878 rubles per patient. However, cumulative treatment costs of R-DA-EPOCH including chemotherapy of the second and further lines and supportive care were 2,896,519 rubles which exceeded those in the R-mNHL-BFM-90 group. Due to its higher efficacy the R-mNHL-BFM-90 immunochemotherapy precludes additional costs associated with both chemotherapy of the second and further lines and supportive care. Conclusion. R-mNHL-BFM-90 as intensive induction block immunochemotherapy for DLBCL patients with poor prognosis is more effective than R-DA-EPOCH and allows to considerably reduce cumulative costs. It is possible due to complete preclusion of the costs of second-line chemotherapy and supportive care including blood component transfusions.

Background. Primary mediastinal (thymic) large B-cell lymphoma (PMBCL) is an aggressive malignant lymphoproliférative disease which accounts for 2-3 % of all non-Hodgkin's lymphomas. In 40 % of PMBCL cases rearrangements of the MHC class II activator, i.e. CIITA gene, are observed. CIITA abnormalities lead to decreasing protein expression and surface expression of MHC class II, which results in lack of adaptive cell immunity targeted at tumor cells. Aim. To assess the rate and spectrum of cytogenetic and molecular genetic abnormalities of CIITA gene in PMBCL patients. Materials & Methods. The study enrolled 37 patients with diagnosed PMBCL: 10 men and 27 women aged 21-61 years (median of 31 years). Sanger sequencing was performed in 36 patients. In 20 patients CIITA/16p13.13 FISH and in 15 patients standard cytogenetic analysis were carried out. Results. In 3 (8.3 %) out of 36 patients the sequencing method detected mutations impairing CIITA gene function, as well as microdeletion in exon 1, deletion and nucleotide substitution in a splice donor site. Multiple somatic variations in intron 1 were identified in 21 (58.3 %) patients: in 11 (52.4 %) cases there were deletions and single nucleotide variants (SNV); the other 10 (47.6 %) patients showed only SNVs. In 13 (61.9 %) out of 21 cases the abnormalities of promoter IV and/or alternative exon 1 were observed. In 5 (25 %) out of 20 patients the FISH assay identified CIITA gene translocation. Standard cytogenetic analysis detected complex karyotype in 7 (46.6 %) out of 15 patients. The comparison of data showed hypermutagenesis in 8 out of 10 patients with FISH-detected chromosome aberrations, and in 3 (37.5 %) of them complex karyotype aberrations were found as well. Conclusion. Molecular genetic methods identified different somatic variations in CIITA gene affecting its functionally important regions, which can be of special interest for further studying the biology of tumors, including PMBCL.

Background. The management of aggressive lymphomas in pregnancy depends on the time of diagnosis and immu-nomorphological variant of tumor. The rarity of aggressive lymphomas in pregnant women, the absence of consistent approaches to the treatment of such patients, the lack of data on physical growth of children as well as the incidence of newborns’ congenital and acquired pathology make this subject of vital importance. Aim. To analyze the treatment results in patients with newly diagnosed aggressive lymphoma at different stages of pregnancy. Materials & Methods. From 1993 to 2020 at the National Research Center for Hematology 74 pregnant women with lymphomas were treated. Aggressive tumors were detected in 17 (23 %) of them: primary mediastinal (thymic) large B-cell lymphoma (п = 14), anaplastic large-cell lymphoma ALK+ (п = 1), high-grade B-cell lymphoma, unspecified (п = 1), and diffuse large B-cell lymphoma (п = 1). The median age of patients was 30 years (range 21-37 years). The median pregnancy stage on the diagnosis of aggressive lymphoma was 21 weeks (range 11-32 weeks). Results. In 1 case on the diagnosis of aggressive lymphoma at 11 weeks gestation dexamethasone 8 mg daily was administered up to the second trimester of pregnancy, afterwards the patient received polychemotherapy. On the diagnosis of aggressive lymphoma in the second (п = 13) and third (п = 2) trimesters of pregnancy the patients received polychemotherapy followed by delivery. In the third trimester of pregnancy delivery was performed with subsequent polychemotherapy in 1 patient. There were born 18 babies (1 pregnancy was multifetal): 8 girls and 10 boys. Conclusion. As a result of the chosen tactics and the work of interdisciplinary team of doctors all patients, who completed the treatment, are followed-up in complete remission. All born babies, despite chemotherapy and perinatal complications, are alive and develop without abnormalities.

Aim. To compare NHL BFM-90 and CHOEP efficacy in adult patients with ALK-positive anaplastic large-cell lymphoma (ALK+ ALCL). Materials & Methods. Within the period from June 2014 to December 2019 the prospective randomized comparative study at the National Research Center for Hematology in Moscow included 23 ALK+ ALCL patients. In one study arm (n = 11) CHOEP was administered, whereas the other one (n = 12) received high-dose chemotherapy (CT) according to NHL BFM-90 protocol. The median age of patients in both arms was 33 and 40 years, respectively. Results. Overall survival (OS) and event-free survival (EFS) within 3 years were 91 % in the arm receiving CHOEP (this protocol was administered to all 11 patients), and 100 % in the arm receiving NHL BFM-90 (complete remission was achieved in all patients). Due to its toxicity NHL BFM-90 was fully implemented in 9 out of 12 patients. The 3-year OS and EFS in the CHOEP and NHL BFM-90 arms are comparable, and the difference between them is not significant. Conclusion. In ALK+ ALCL treatment high-dose CT according to NHL BFM-90 protocol has no advantage in terms of the 3-year OS and EFS compared to less toxic regimen CHOEP. A larger sample of patients is required to achieve significant results, which will further lead to a final judgement on feasibility of high-dose regimens in the treatment of adult patients with ALK+ ALCL.

Aim of the issue was to determine indications for intratecal chemotherapy drugs administration to prevent relapse of diffuse large B-cell lymphoma (DLBCL) with central nervous system (CNS) involvement.Since January 2009 to December 2018 102 patients with primary nodal DLBCL over 18 years old were treated in the National Research Center for Hematology, Moscow, Russian Federation. Diagnosis were established in all cases according to histological and immunohistochemical studies which made it possible to exclude the transformation of mature B-cell lymphoma into DLBCL.Isolated leptomeningeal involvement of CNS in the debut of the disease was detected in 1 (0.98%) out of 102 patients with DLBCL. Focal brain tissue involvement was not detected in any patient. More than half of the patients (54%) had a high risk of disease recurrence or progression with CNS involvement: in 8 (7.8%) patients had kidney/adrenal involvement, in the same proportion - bone marrow involvement, paranasal sinuses involvement - in 5 (4.9 %), epidural space - in 7 (6.9%) and breast - in 5 (4.9%) of patients. In 82 (80%) patients, a non-GCB (postgerminal differentiation of B-cell analog) molecular subtype of DLBCL was determined.The introduction of chemotherapy drugs into the spinal canal is recommended in isolated cases of leptomeningeal involvement of CNS at the time of DLBCL onset and is carried out according to standard recommendations. Prevention of relapse with involvement of central nervous system using intratecal chemotherapy in patients with nodal form of DLBCL is not indicated due to the absence of cases with disease progression or recurrence into CNS when patients were treated with R-m-NHL-BFM-90, R-DA-EPOCH and R-CHOP protocol.Целью работы является определение показаний к проведению профилактики рецидива диффузной В-клеточной крупноклеточной лимфомы (ДВККЛ) с вовлечением центральной нервной системы (ЦНС) путем введения химиопрепаратов в спинномозговой канал. Материалы и методы. С января 2009 по декабрь 2018 г. в ФГБУ «НМИЦ гематологии» Минздрава России проводилось лечение 102 больным первичной нодальной ДВККЛ старше 18 лет. Всем больным проводили гистологическое и иммуногистохимическое исследования, позволившие исключить трансформацию зрелоклеточной лимфомы в ДВККЛ. Результаты. Вовлечение оболочек головного мозга/нейролейкемия в дебюте заболевания диагностировано у 1 (0,98%) больного. Очагового вовлечения вещества головного мозга не выявлено ни у одного пациента. Более половины больных (54%) имели высокий риск развития рецидива или прогрессии с вовлечением ЦНС: у 8 (7,8%) больных отмечалось вовлечение почек/надпочечников, в таком же проценте - костного мозга, параназальных синусов - у 5 (4,9%), эпидурального пространства - у 7 (6,9%) и молочных желез - у 5 (4,9%) больных. У 82 (80,1%) пациентов определен постгерминальный (из В-клеток постгерминальной дифференцировки - non-GCB) молекулярный подтип ДВККЛ. Заключение. Введение химиопрепаратов в спинномозговой канал показано в единичных случаях, когда в момент манифестации ДВККЛ у больных выявляется вовлечение оболочек/нейролейкемия, и проводится согласно стандартным рекомендациям. Профилактическое введение цитостатических препаратов в спинномозговой канал не показано ввиду отсутствия рецидивов ДВККЛ с вовлечением ЦНС у больных, проходивших лечение как по протоколу m-NHL-BFM-90, так и R-CHOP, R-DA-EPOCH.

Aim of the issue was to compare clinical characteristics and treatment results of patients with follicular lymphoma (FL) with translocations involving loci of c-MYC/8q24, BCL2/18q21 and/or BCL6/3q27 genes and patients with high - grade B-cell lymphoma [High - grade B-cell lymphoma (HGBL), double - hit (DH)]. Materials and methods. Since 2004 to 2017 years in National Research Center for Hematology 12 patients with high - grade B-cell lymphoma double - hit (HGBL DH) and 6 FL patients with translocations involving c-MYC and BCL2 and/or BCL6 had been treated. We performed a comparative analysis of clinical characterisctics in both groups. As primary endpoints was assessed frequency of complete remission (CR) or progressive disease (PD); as secondary endpoints - overall (OS) and event - free survival (EFS). Results. 5 patients with HGBL DH had c-MYC/BCL6, 7 - c-MYC/BCL2 rearrangements; 2 patients with FL had c-MYC/BCL2, 3 - c-MYC/BCL6, 1 - c-MYC/BCL2/BCL6 rearrangements. FL was represented by grade 3A in 2, grade 3B - in 4 cases, 3 of them had large - cell transformation. In HGBL DH and FL patients had no significant differences in clinical characteristics. The majority of patients had a widespread tumour, increased LDH activity, high frequency of extranodal and bone marrow involvement. Ki-67 expression level was lower in patients with FL (p.Цель работы: сопоставить клинические характеристики и результаты терапии больных фолликулярной лимфомой (ФЛ) с транслокациями с участием локусов генов с-MYC/8q24, BCL2/18q21 и/или BCL6/3q27 и больных В-клеточной лимфомой высокой степени злокачественности [high-grade B-cell lymphoma (HGBL)], double-hit (DH). Материалы и методы. С 2004 по 2017 г. в ФГБУ «НМИЦ гематологии» Минздрава России наблюдались 12 больных с диагнозом HGBL DH и 6 больных ФЛ с транслокациями с участием локусов генов c-MYC, BCL2 и/или BCL6. Мы выполнили сравнительный ретроспективный анализ клинических характеристик в обеих группах пациентов. В качестве первичных конечных точек оценена частота полных ремиссий (ПР) и прогрессирования заболевания (Прогр), в качестве вторичных - общая (ОВ) и бессобытийная выживаемость (БСВ). Результаты. У 5 больных с HGBL DH выявлены транслокации с участием локусов генов c-MYC/BCL6, у 7 - c-MYC/BCL2; у 2 с диагнозом ФЛ - c-MYC/BCL2, у 3 - c-MYC/BCL6, у 1 - c-MYC/BCL2/BCL6. Цитологический тип ФЛ представлен ФЛ 3А с бластоидной морфологической картиной в 2 случаях, ФЛ 3В в 4 случаях, 3 из которых имели признаки гистологической трансформации. Больные HGBL DH и ФЛ не имели значимых различий по клиническим характеристикам. Так, большинство из них имели распространенную стадию заболевания, повышенную активность лактатдегидрогеназы, высокую частоту вовлечения экстранодальных областей и костного мозга. Уровень пролиферативной активности Ki-67 был значимо выше у больных HGBL DH (p.

Background. Primary mediastinal (thymic) large B-cell lymphoma (PMBCL) is a malignant tumor with large atypical lymphoid cells expressing post-germinal differentiation markers. Rearrangements of immunoglobulin genes in PMBCL are revealed in 30-65 % of cases. Immunoglobulin molecules, however, are expressed neither on the surface, nor in cytoplasm of tumor cells. Aim. To assess cell clonality rate on the basis of rearrangements of immunoglobulin heavy/light chain genes; to determine rearrangement stability at the time of relapse development; to study the range of rearrangements and clonal relationship with primary tumor in metachronous development of mediastinal gray zone lymphoma. Materials & Methods. The assessment of rearrangements of immunoglobulin heavy/light chain genes was based on molecular analysis of 29 primary tumor biopsies and 4 tissue samples with histologically and immunohistochemically verified relapses or metachronous lymphoma development. Results. In 16 (55.2 %) out of 29 cases a rearrangement of immunoglobulin heavy chain genes was reported, in 7 (24.1 %) cases a rearrangement of light chain genes was identified, in 6 (20.7 %) cases no rearrangements of immunoglobulin heavy/light chain genes were found. On the basis of immunoglobulin gene analysis in 2 patients with early relapse a tumor clone was detected that was identical with the one identified at the onset of the disease. In 2 patients with complete remission a metachronous development of mediastinal gray zone lymphoma was reported, whereas molecular genetic analysis revealed a change/disappearance of initial clonal rearrangements of immunoglobulin genes. Conclusion. Total detection rate of B-cell clonality in PMB-CL was 79.3 %. Molecular genetic analysis confirmed that initial clonal rearrangements of immunoglobulin genes were preserved in early relapses, and invalidated tumor clonal relationship in a metachronous development of mediastinal gray zone lymphoma.

Background. TP53 gene mutations impede cell apoptosis and lead to additional oncogenic events contributing to tumor progression. Aim. To assess TP53 gene mutation rate in patients with high-grade B-cell lymphoma double-hit (HGBCL DH) and not otherwise specified (HGBL NOS); to analyse its relationship with disease prognosis. Materials & Methods. Retrospective materials from medical data of 10 HGBL DH and 26 HGBL NOS patients were analyzed. Median follow-up was 26.5 months (range 0.6-160.9 months). Selection was based on the presence of available biological materials (paraffin blocks) for Sanger sequencing of TP53 gene from exon 5 to exon 8 (encoding DNA-binding domain of TP53 gene). FISH analysis of the tumor was performed in all patients to identify translocations involving c-MYC/8q24, BCL2/18q21, and BCL6/3q27 gene locus. To analyze differences between groups χ2 and Mann-Whitney tests were applied. Univariate event analysis (Kaplan-Meier and log-rank tests) and Cox regression analysis were used to assess the influence of molecular markers on the disease prognosis. Results. TP53 gene mutations in lymphoma cells were found in 13 (36 %) out of 36 patients, 10 (77 %) out of 13 mutations were pathogenic. In 8 out of 10 patients with TP53 mutations c-MYC/8q24 gene translocation was identified. Groups with wild (TP53-WT) and mutant (TP53-MUT) types of TP53 gene were similar in terms of main clinical characteristics. Patients with TP53-MUT in tumor cells showed worse 3-year overall survival (OS) compared with the group without TP53-MUT (30 % vs. 73 %; p = 0.026) as well as higher probability of disease progression in the period of 3 years (66 % vs. 15 %; p = 0.004). In multivariate analysis significant OS predictor was the presence of TP53 mutation (p = 0.006). Relapse/ progression probability was higher in combined cases of TP53 mutation and translocation involving c-MYC gene locus (p = 0.0003). Conclusion. Translocation involving c-MYC gene along with TP53 gene mutation in tumor cells can serve as a criterion for dividing HGBL DH and HGBL NOS patients into different lymphoma relapse/progression risk groups.

Analysis of microsatellite instability (MSI) is a routine study in the diagnostics of solid malignancies. The standard for determining MSI is a pentaplex PCR panel of mononucleotide repeats: NR-21, NR-24, NR-27, BAT-25, BAT-26. The presence of MSI is established based on differences in the length of markers in the tumor tissue and in the control, but due to the quasimonomorphic nature of standard mononucleotide loci the use of a control sample is not necessary in the diagnosis of MSI-positive solid tumors. The significance of the MSI phenomenon in oncohematology has not been established. This paper presents the results of a study of MSI in B-cell lymphomas: follicular lymphoma (FL), diffuse large B-cell lymphoma (DLBCL), high-grade B-cell lymphoma (HGBL). We have shown that aberrations of mononucleotide markers occur in these diseases, but the nature of the changes does not correspond to the classical MSI in solid neoplasms. This fact requires further study of the pathogenesis of such genetic disorders. Due to the possibility of ambiguous interpretation of the results of the MSI study for previously uncharacterized diseases, strict compliance with the methodology of parallel analysis of the tumor tissue and the control sample is mandatory.

Background. Grade 3 follicular lymphoma (FL) is a heterogenetic group of tumors. The selections of patients with similar characteristics of the tumor process is important for classification 3 grade forms of FL and risk stratification, as well as for the development of new therapeutic approaches. Different morphological, immunohistochemical and cytogenetical characteristics of the tumor result in different clinical forms of the disease. Aim. To describe the clinical, morphological, immunohistochemical and cytogenetical characteristics of grade 3 FL and evaluate their prognostic value for R-CHOP-21 chemotherapy. Materials & Methods. We performed retrospective and prospective analysis of morphological, immunohistochemical and genetical characteristics of 93 primary patients with grade 3 FL (21-78 years, median 53 years, women to men - 1:1.4) admitted to National Medical Hematology Research Center from years 2001 to 2016. Morphological and immunohistochemical assessment of the affected lymph nodes and bone marrow biopsy material was performed. Data obtained from the standard cytogenetic and FISH assessment were compared to identify the BCL2 rearrangement. Results. We proposed an algorithm for differential diagnosis of the 2 types of grade 3 FL: de novo FL (n = 22) and transformed FL (n = 21). De novo grade 3 FL had the immunophenotype of CD10- in 19 (86 %) cases, MUM1++ (monomorphically) in 19 (90 %), and BCL-2 in 5 (22 %). It was characterized by the absence of the BCL2 rearrangement (n = 22, 100 %) and bone marrow involvement (n = 14, 67 %) and/or bone marrow involvement (n = 7, 100 %). Third grade FL transformed from grades 1 or 2 had was CD10+ (n = 19, 90 %), MUM1+ (heterogeneously, n = 16, 76 %) or MUM1-(n = 4, 19 %), BCL-2+ (n = 20, 95 %) and had BCL2 rearrangement (n = 19, 90 %). Small cell bone marrow involvement was observed in 71 % of cases, whereas large cell involvement was seen predominantly in de novo FL (p = 0.06). The analysis showed that 5-year relapse-free survival in patients with grade 3 de novo FL after R-CHOP-21 therapy was 87 % compared to 16 % with transformed FL (p = 0.06) for the median 41 months of follow up. Conclusion. We described the morphological, immunohistochemical and cytogenetical characteristics of grade 3 de novo FL and grade 3 FL, transformed from grades 1 or 2. The described variants show different sensitivity to immunochemotherapy.

Primary mediastinal large B-cell lymphoma (PMBCL) is a distinct type of large B-cell lymphoma. In this type of the disease, the neoplastic process is located in the anterior and superior mediastinum, frequently with compression of the superior vena cava and with tumor invasion into the adjacent organs and tissues: the pericardium, lung, pleura, etc. Despite the fact that in PMBCL progression, there may be involvement of extranodal organs, such as the kidney, adrenal glands, liver, and central nervous system, bone marrow (BM) injury is generally absent. Since BM injury in patients with diffuse large B-cell lymphoma is an independent poor prognostic indicator, there is reason to believe that BM involvement in PMBCL affects the prognosis. These cases may need intensified induction therapy followed by autologous hematopoietic stem cell transplantation; and BM injury should be monitored during the therapy. The paper gives reports of clinical cases of bone marrow involvement in 2 PMBCL patients treated at the National Research Center for Hematology, Ministry of Health of the Russian Federation.Первичная медиастинальная В-клеточная крупноклеточная лимфома (ПМВККЛ) является отдельной формой В-клеточной крупноклеточной лимфомы. При этой форме заболевания опухолевый процесс локализуется в переднем и верхнем средостении, нередко со сдавливанием верхней полой вены и прорастанием опухоли в прилежащие органы и ткани: в перикард, легкие, плевру и т.д. Несмотря на то что при прогрессии ПМВККЛ возможно вовлечение таких экстранодальных органов, как почки, надпочечники, печень и центральная нервная система, поражение костного мозга (КМ), как правило, отсутствует. Поскольку поражение КМ у больных ДВККЛ является независимым признаком неблагоприятного прогноза, есть основания предположить, что вовлечение КМ при ПМВККЛ влияет на прогноз. В подобных случаях, возможно, необходима интенсификация индукционной терапии с последующей трансплантацией аутологичных стволовых кроветворных клеток, а в ходе терапии следует проводить мониторинг поражения КМ. В статье представлены описания клинических случаев с поражением КМ у 2 больных ПМВККЛ в ФГБУ ГНЦ МЗ РФ.

To estimate the number of early progenitors of bone marrow (BM) hematopoiesis in patients with diffuse large B-cell lymphoma (DLBCL) in the late period after high-dose chemotherapy (HDCT) according to the mNHL-BFM-90 program.The investigators analyzed the results of BM immunophenotypic and histological studies in 40 patients (median age, 57 years) with DLBCL who received HDCT according to the mNHL-BFM-90 program at the Hematology Research Center (HRC), Ministry of Health of the Russian Federation (MHRF), in the period 2002 to 2009. A comparison group consisted of 19 patients (median age, 70 years) treated according to the CHOP/R-CHOP program at HRC, MHRF, in the same period. The median follow-up period was 6 years. The results of BM examination were analyzed before and 5-10 years after the end of HDCT. Immunophenotypic study determined the number of early CD34+ hematopoietic progenitors. BM cellularity, the size of erythroid, granulocytic and megakaryocytic lineages, their ratio, the presence of dysplasia signs, and secondary stromal changes were histologically determined. The BM toxic injury signs found for the first time were evaluated as manifestations of late myelotoxicity.At 5-to-10-year follow-ups after the end of HDCT according to the mNHL-BFM-90 program, the patients showed a smaller number of early CD34+ progenitors of BM hematopoiesis in 31 (78%) cases than those treated according to the CHOP/R-CHOP-21 program (n=8 (2%)) (p=0.005). Myelopoiesis with decreased CD34+ cell count was characterized by hypocellularity in 8 (26%) patients (p=0.07), the narrowing of megakaryocytic lineage in 14 (45%) (p=0.006), erythroid one in 7 (23%) (p=0.01), and granulocytic one in 8 (26%) (p=0.92), pronounced secondary stromal changes in 15 (48%) (p=0.03), and grade 1 thrombocytopenia in 13 (42%); p=0.02).There is evidence that the number of early CD34+ progenitors of BM hematopoiesis decreased in patients with DLBCL in the late period after HDCT. The investigation shows the relationship of the reduction in the number of early CD34+ progenitors of BM hematopoiesis in the late follow-up period to the presence of pronounced secondary changes in the BM stroma (p=0.02). There was no statistically significant relationship of the decreased number of CD34+ cells to the age younger or older than 60 years, to the period after the end of chemotherapy, to gender or presence of specific BM injury.Цель исследования. Оценка количества ранних предшественников кроветворения в костном мозге (КМ) больных диффузной В-крупноклеточной лимфомой (ДВККЛ) в отдаленном периоде после высокодозной химиотерапии (ВДХТ) по программе mNHL-BFM-90. Материалы и методы. Проанализированы результаты иммунофенотипического и гистологического исследований КМ 40 больных ДBККЛ, которым выполнена ВДХТ по программе mNHL-BFM-90 в ГНЦ МЗ РФ в период с 2002 по 2009 г., медиана возраста 57 лет. Группу сравнения составили 19 пациентов, которым проведена терапия по программе СНОР/R-СНОР в ГНЦ МЗ РФ в тот же период, медиана возраста 70 лет. Медиана срока наблюдения после окончания лечения 6 лет. Анализировали результаты исследования КМ до начала ВДХТ и через 5-10 лет после окончания лечения. При иммунофенотипическом исследовании определяли количество ранних предшественников кроветворения CD34+. При гистологическом исследовании определяли клеточность КМ, величину эритроидного, гранулоцитарного и мегакариоцитарного ростков, их соотношение, наличие признаков дисплазии, а также вторичные изменения стромы. В качестве проявлений отдаленной миелотоксичности оценивали впервые выявленные признаки токсического поражения КМ. Результаты. У больных ДВККЛ после высокодозной терапии по программе mNHL-BFM-90 при сроке наблюдения от 5 до 10 лет после окончания химиотерапии выявлено снижение количества ранних предшественников кроветворения CD34+ в КМ в 31 (78%) случае, в отличие от 8 (42%) больных после терапии СНОР/R-СНОР-21 (р=0,005). Миелопоэз со сниженным количеством клеток CD34+ характеризовался гипоклеточностью у 8 (26%; р=0,07), сужением мегакариоцитарного ростка у 14 (45%; р=0,006), эритроидного у 7 (23%; р=0,01) и гранулоцитарного у 8 (26%; р=0,92), выраженными вторичными изменениями стромы у 15 (48%; р=0,03), тромбоцитопенией I степени у 13 (42%; р=0,02) больных. Заключение. Доказано уменьшение количества ранних предшественников кроветворения CD34+ в КМ у больных ДВККЛ в отдаленном периоде после ВДХТ. Показана зависимость снижения количества ранних предшественников кроветворения CD34+ в КМ в отдаленном периоде наблюдений от наличия выраженных вторичных изменений стромы КМ (р=0,02). Статистически значимой зависимости уменьшения количества клеток CD34+ от возраста моложе или старше 60 лет, срока после окончания химиотерапии, пола, наличия специфического поражения КМ не выявлено.Цель исследования. Оценка количества ранних предшественников кроветворения в костном мозге (КМ) больных диффузной В-крупноклеточной лимфомой (ДВККЛ) в отдаленном периоде после высокодозной химиотерапии (ВДХТ) по программе mNHL-BFM-90. Материалы и методы. Проанализированы результаты иммунофенотипического и гистологического исследований КМ 40 больных ДBККЛ, которым выполнена ВДХТ по программе mNHL-BFM-90 в ГНЦ МЗ РФ в период с 2002 по 2009 г., медиана возраста 57 лет. Группу сравнения составили 19 пациентов, которым проведена терапия по программе СНОР/R-СНОР в ГНЦ МЗ РФ в тот же период, медиана возраста 70 лет. Медиана срока наблюдения после окончания лечения 6 лет. Анализировали результаты исследования КМ до начала ВДХТ и через 5—10 лет после окончания лечения. При иммунофенотипическом исследовании определяли количество ранних предшественников кроветворения CD34+. При гистологическом исследовании определяли клеточность КМ, величину эритроидного, гранулоцитарного и мегакариоцитарного ростков, их соотношение, наличие признаков дисплазии, а также вторичные изменения стромы. В качестве проявлений отдаленной миелотоксичности оценивали впервые выявленные признаки токсического поражения КМ. Результаты. У больных ДВККЛ после высокодозной терапии по программе mNHL-BFM-90 при сроке наблюдения от 5 до 10 лет после окончания химиотерапии выявлено снижение количества ранних предшественников кроветворения CD34+ в КМ в 31 (78%) случае, в отличие от 8 (42%) больных после терапии СНОР/R-СНОР-21 (р=0,005). Миелопоэз со сниженным количеством клеток CD34+ характеризовался гипоклеточностью у 8 (26%; р=0,07), сужением мегакариоцитарного ростка у 14 (45%; р=0,006), эритроидного у 7 (23%; р=0,01) и гранулоцитарного у 8 (26%; р=0,92), выраженными вторичными изменениями стромы у 15 (48%; р=0,03), тромбоцитопенией I степени у 13 (42%; р=0,02) больных. Заключение. Доказано уменьшение количества ранних предшественников кроветворения CD34+ в КМ у больных ДВККЛ в отдаленном периоде после ВДХТ. Показана зависимость снижения количества ранних предшественников кроветворения CD34+ в КМ в отдаленном периоде наблюдений от наличия выраженных вторичных изменений стромы КМ (р=0,02). Статистически значимой зависимости уменьшения количества клеток CD34+ от возраста моложе или старше 60 лет, срока после окончания химиотерапии, пола, наличия специфического поражения КМ не выявлено.

Context: Lymphoproliferative diseases (LPD) occurring during pregnancy present the treating physician with unique diagnostic and therapeutic challenges, such as aiming to achieve maternal cure without impairing fetal health, growth, and survival. Design: Analysis of the treatment results of patients with aggressive LPD diagnosed at different stages of pregnancy. Setting: From 2009 to 2020, 16 pregnant women with aggressive LPD were treated in the National Research Center for Hematology. Patients or other participants: Primary mediastinal large B-cell lymphoma (n = 13), ALK + anaplastic large-cell lymphoma (n = 1), high-grade B-cell lymphoma (n = 1), diffuse large B-cell lymphoma (n = 1). The median age was 30 (21–37) years. Results: When diagnosing LPD in the second trimester of pregnancy (TP), chemotherapy (CT) was performed followed by delivery (n = 13), when diagnosed in III TP, depending on gestational age, either delivery followed by CT (n = 1), or performed CT and delivery at a later gestational age (n = 2). All patients at the time of diagnosis had indications for CT. Treatment of patients was performed according to the protocol: DA-EPOCH±R - 9, VACOP-B - 5, CHOEP - 1, CHOP - 1. The gestational age at the time of delivery after CT was 34 (25–36) weeks. Currently, all patients are alive and are in complete remission of the disease. The median follow-up was 41 (1–246) months. Seventeen children were born (1 pregnancy was multiple): they are alive, healthy, and without developmental disabilities. The age of children at the time of data analysis ranges from 1 month to 10 years. Conclusions: As a result of the chosen management tactics, all patients are alive, in complete remission of the disease, and the children are practically healthy.

Mediastinal gray-zone lymphoma (MGZL, lymphoma with features intermediate between classical Hodgkin lymphoma and diffuse large B-cell lymphoma) was declared as a separate entity in WHO classification of Tumors of Haematopoetic and Lymphoid Tissues in 2008 and 2017 years. Despite of similar pathomorphological characteristics between primary mediastinal B-cell lymphoma and Hodgkin lymphoma, clinical features and optimal therapeutic approach to MGZL are not clearly defined. Usually MGZL manifests with mediastinal lymphadenopathy, although extranodal lesions often occur (grey-zone lymphoma, GZL). Patients with MGZL have unfavorable prognosis, taking into account high rate of relapse. This article describes two cases of MGZL. First case manifested by arrhythmias due to primary heart involvement. In spite of cardiac failure antracycline-containing chemotherapy (6 courses of R-DA-EPOCH) it allowed to achieve a complete remission and resolving of arrhythmias. Second case was represented by metachronous tumors: primary mediastinal B-cell lymphoma at the time of disease onset and classical Hodgkin lymphoma, NS II, diagnosed after disease progression. Thus, we demonstrated two examples of MGZL that differ by clinical manifestation, response to chemotherapy, which emphasizes an importance of pathogenesis studying, and using of new therapeutic approaches.

Purpose. The identifying of the clinical, morphological, immunophenotypic and cytogenetic features of double-hit and double-expressor lymphomas as well as the development of optimal tactics of treatment and evaluation of efficiency of the ASCT were the main purposes of the study. Material and methods. The study included 85 patients: 45 males and 40 females, the median was 46,6 years (15-73). 62 patients with DLBCL and 23 with BCLu were given BL-M-04/ m-NHL-BFM-90 regimes in Hematological Scientific Center and Municipal Clinical Hospital №52, 2001-2015. Results. Stage I was diagnosed in 5 (6%) patients, II - in 13 (15%) patients, III - in 7 (8%) patients, IV - in 60 (71%) patients. Double-hit lymphoma was diagnosed in 6 (7%) patients (5 MYC+/BCL2+, 1 MYC+/BCL6+), single-hit lymphoma was diagnosed in 6 patients. The translocation was detected in 5 of 12 cases t(8;14)(q24;q32), in 1 case of 12 - t(2;8)(p21;q24), in 2 cases of 12 - t(8;22)(q24;q11) (restructuring c-MYC and IGL gene loci have been identified), in 3 cases of 12 - restructuring of c-MYC with non-IG partner [translocation was absent t(8;14)(q24;q32) and restructuring IGK and IGL gene loci], the partner was not identified in one case of translocation with c-MYC gene [restructuring c-MYC gene loci was identified and translocation was absent t(8;14(q24;q32)]. 8 factors had prognostic significance in relation to OS: nosology - BCLu vs DLBCL (p=0.007; RR=4.5), bone marrow lesion (p=0.0004; RR=7.9), B-symptoms (p=0.0002; RR=8.4), BCL2 expression (I=0.01; RR=4.4), MYC/BCL2 coexpression (p=0.017; RR=0.08), double-hit or double-expressor lymphomas (p=0.009; RR=15.1), the absence of rituximab in therapy (p=0.03; RR=0.3), unfulfillment of ASCT (p=0.003; RR=0.06). Two factors were significant in relation to probability of development of recurrence/progression: double-hit or double-expressor lymphomas (p=0.0005; RR=4.0) and IPI (p=0.03; RR=4.9). Conclusions. Thus, complex diagnostic includes the antibody to the BCL2 (clone 124, Dako) and MYC (clone Y69, Epitomics), and the conduct of standard cytogenetic analysis (SCA) and FISH to detect rearrangements of MYC, BCL2, and BCL6 genes in addition to standard immunohistochemical panel for aggressive B-cell lymphomas. It allows not only to establish the correct diagnosis (DLBCL, BCLu or LB), but also to detect the double-hit and double-expressor lymphomas, which require intensive induction with rituximab and high-consolidation ASCT in first remission.

Background: Anaplastic large cell ALK-positive lymphoma (ALCL ALK+) is a lymphoproliferative disease of the “young” age. ALCL ALK+ has a favourable prognosis. A failure in therapy (relapse) is diagnosed within the first 1.5-2 years of the follow up period. Aim: The aim of the present study was to compare the effectiveness of the NHL BFM-90 and CHOEP protocols intended for the treatment of adult patients presenting with the ALCL ALK+. Materials and methods: This prospective comparative randomized study was carried out based at the National Medical Research Centre of Hematology, Moscow, during the period from June 2014 to December 2019. It involved 23 patients with the ALCL ALK+ disease randomly allocated to two groups. One of them included 11 patients given a CHOEP, the other consisted of 12 patients treated using a high-dose CT in accordance with the NHL BFM-90 protocol (age medians 33 and 40 years respectively). Results: The 3-year OS and EFS rates among the patients receiving CHOEP was 91%, with all the members of this study group having completed the treatment regimen. All patients comprising the latter (NHL BFM-90) group achieved complete remission. However, only 9 of the 12 patients fully complied with the treatment protocol because of its toxic effects. It can be inferred that the two study groups are comparable in terms of the 3-year OS and EFS rates among the patients treated in accordance with the CHOEP and NHL BFM-90 programs, the differences between them being insignificant. Conclusion: High-dose CT based on the NHL BFM-90 protocol has no advantages over the less toxic CHOEP regimen as far as the treatment of the ALCL ALK+ is concerned (if the efficacy of both modalities is estimated in terms of the 3-year OS and EFS rates). However, additional studies with larger patient samples are needed to obtain statistically significant results before a definitive conclusion can be made as regards clinical feasibility of the use of high-dose therapeutic regimens for the treatment of adult patients presenting with the ALCL ALK+ disease.

Context: Genetic instability is one of the characteristic features of tumor cells and is associated with tumor progression and transformation. An integral measure of genetic instability could be assessed through STR (short tandem repeats) analysis in terms of MSI (microsatellite instability) and EMAST (elevated microsatellite alteration at selected tetranucleotides). MSI and EMAST occur as a result of DNA repair complex malfunction in tumor cells. The study presents MSI and EMAST data in patients with non-Hodgkin B-cell lymphomas: follicular lymphoma (FL) and diffuse large B-cell lymphoma (DLBCL). Objective: To determine the frequency of MSI and EMAST as predictors of the clinical course of the disease at the early stages of diagnosis in patients with FL and DLBCL. Design: FL patients (n=7) and DLBCL patients (n=5) who followed up at the National Research Center for Hematology were included in the study after receiving their informed consent. DNA samples from tumor and control DNA samples from healthy somatic cells of the same patient were isolated according to a standard protocol. Genetic instability was assessed by multiplex PCR of STR loci with COrDIS Plus and COrDIS MSI kits (Gordiz Ltd, Russia). The fragment analysis was performed on an ABI3130 Genetic Analyzer. Results: The observed frequency of MSI and EMAST in FL was 15% (1/7) and 28% (2/7); in DLBCL, 0% (0/5) and 20% (1/5), respectively. STR profiles also revealed allelic imbalance due to loss of heterozygosity (LOH) with a high incidence: 71% (5/7) and 60% (3/5) in FL and DLBCL, respectively. LOH is an independent form of genetic instability and probably associated with recombination complex disruption in tumor cells. LOH reflects the loss of a significant amount of genetic material from a single chromosome via deletions or uniparental disomy. Conclusions: Statistically significant measures could not be reported here due to the small sample size. However, the findings show that STR instability is not a rare event in non-Hodgkin B-cell lymphomas; therefore, further detailed analysis of extended patient cohorts to determine its clinical significance is required.

to evaluate the efficiency of the R-DA-EPOCH-21 + R-DHAP protocol and autologous hematopoietic stem cell transplantation (a BEAM conditioning mode) in first-line therapy for primary mediastinal large B-cell lymphoma (PMBCL).In 2013 to 2016, the investigation enrolled 57 patients with newly diagnosed PMBCL (according to the 2008 WHO criteria). The results were analyzed in 40 patients who had completed their treatment.All the 40 patients (14 men and 26 women) (median age, 27 years (19 to 67 years)) received 6 cycles of polychemotherapy (PCT) in accordance with the R-DA-EPOCH-21 regimen. After induction PCT cycles, 32/40 (80%) patients achieved complete remission. Partial remission was stated in 8/40 (20%) patients who had further 2 cycles of chemotherapy using the R-DHAP program and autologous hematopoietic stem cell transplantation (a BEAM conditioning mode). Two-year overall and relapse-free survival rates were 100% and 96%, respectively; the median follow-up was 17 months.The R-DA-EPOCH regimen allows complete remission in 80% of the cases and two-year survival in 100%. If there are unfavorable factors at onset and in partial remission, it is appropriate to intensify treatment at early stages, by using high-dose chemotherapy and autologous hematopoietic stem transplantation.Цель исследования. Оценить эффективность протокола R-DA-EPOCH 21 + R-DHAP + трансплантация аутологичных гемопоэтических стволовых клеток крови (режим кондиционирования BEAM) в терапии первой линии при первичной медиастинальной В-крупноклеточной лимфоме (ПМВЛ). Материалы и методы. С 2013 по 2016 г. в исследование включили 57 больных с впервые установленным диагнозом ПМВЛ (согласно критериям ВОЗ, 2008 г.). Результаты проанализированы у 40 пациентов, завершивших лечение. Результаты. Всем 40 пациентам (14 мужчин и 26 женщин, медиана возраста 27 лет, от 19 до 67 лет) выполнили 6 циклов полихимиотерапии (ПХТ) по схеме R-DA-EPOCH-21. После индукционных курсов ПХТ полную ремиссию заболевания достигли 32/40 (80%) больных. Частичную ремиссию заболевания констатировали у 8/40 (20%) пациентов и в дальнейшем им выполнили 2 цикла химиотерапии по программе R-DHAP и трансплантацию аутологичных гемопоэтических стволовых клеток крови (режим кондиционирования ВЕАМ). Двухлетняя общая выживаемость и безрецидивная выживаемость составили 100 и 96% соответственно, медиана наблюдения 17 мес. Заключение. Выполнение схемы R-DA-EPOCH позволяет достигнуть полной ремиссии заболевания в 80% случаев и вероятность прожить 2 года 100%. При наличии факторов неблагоприятного прогноза в дебюте и достижении частичной ремиссии после индукционных курсов целесообразна интенсификация лечения на ранних этапах с использованием высокодозной химиотерапии и трансплантации аутологичных гемопоэтических стволовых клеток крови.

Splenic marginal zone B-cell lymphoma (SMZBCL) is a rare non-Hodgkin B-cell lymphoma that presents with morphologically mature lymphoid cells corresponding in their immunological characteristics to secondary follicular marginal zone lymphocytes. It is clinically characterized by splenomegaly, moderate lymphocytosis, usually focal bone marrow lesion, sometimes moderate of monoclonal immunoglobulin in the serum (generally IgM or IgG) and/or urea, and a relatively benign course. Leishmaniasis is a transmissible natural focal infectious endemic disease that has a great diversity of clinical manifestations. The authors describe Russia's first case of SMZBCL concurrent with visceral leishmaniasis in a 52-year-old female patient admitted to a hematology hospital with weakness, splenomegaly, and lymphadenopathy. The simultaneous detection of lymphoma and leishmaniasis in the same biopsy specimen is extremely rare. Visceral leishmaniasis should be borne in mind as an opportunistic infection in patients with malignancies, particularly in immunocompromised persons who live or have stayed in the endemic areas.Селезеночная В-клеточная лимфома из клеток маргинальной зоны (СЛКМЗ) - редкая В-клеточная неходжкинская лимфома (НХЛ), которая представлена морфологически зрелыми лимфоидными клетками, по своим иммунологическим характеристикам соответствующим лимфоцитам маргинальной зоны вторичного фолликула. Клинически характеризуется спленомегалией, умеренным лимфатическим лимфоцитозом, обычно очаговым поражением костного мозга, иногда умеренной секрецией моноклонального иммуноглобулина в сыворотке крови (обычно IgM или IgG) и/или моче и относительно доброкачественным течением. Лейшманиоз - трансмиссивное природно-очаговое инфекционное эндемическое заболевание, имеющее разнообразные клинические проявления. Мы приводим первый случай в России сочетания СЛКМЗ с висцеральным лейшманиозом у больной 52 лет, госпитализированной в гематологический стационар по поводу слабости, спленомегалии и лимфаденопатии. Одновременное обнаружение лимфомы и лейшманий в одном и том же биоптате крайне редко. Висцеральный лейшманиоз следует иметь в виду как оппортунистическую инфекцию у больных со злокачественными новообразованиями, особенно с ослабленным иммунитетом, проживающих или побывавших в эндемических областях.

The stromal microenvironment regulating hematopoiesis in patients with hematologic malignancies undergoes significant alterations. The changes in the concentration of colony-forming fibroblast units (CFU-F) in the bone marrow (BM) and disruption in the functioning of multipotent mesenchymal stromal cells (MSCs) are shown in many studies for patients with acute leukemia. Often it is not possible to distinguish the cause of changes in stromal progenitor cells after treatment: interaction with tumor cells or the effects of therapy. Most of the patients with diffuse large B-cell lymphoma (DLBCL) do not have BM involvement. It was assumed that the properties of MSCs in these patients were not changed, and so this could be an attractive model for investigation the effect of antitumor drugs on human BM stromal microenvironment. The aim of the study was to compare the properties of MSCs in patients with DLBCL in the onset of the disease and a month after the end of therapy. Methods The study included 20 patients with DLBCL (11 male, 9 female) aged 42-60 years in the onset of the disease and a month after the end of treatment. 3-5 ml of BM were collected during diagnostic punctures after informed consent. MSCs and CFU-F were cultured by standard methods. The total MSCs production, the doubling-population level per day, the concentration of CFU-F, the relative gene expression level (REL) in MSC by real-time PCR and the mean fluorescence intensity (MFI) by flow cytometry were analyzed. The control group included 31 donors of the corresponding age. The analysis of MSCs secretome was carried out using the LC-MS/MS analysis TripleTOF 5600+ mass spectrometer with a NanoSpray III ion source coupled to a NanoLC Ultra 2D+ nano-HPLC System. Results The total cell production for 4 passages in primary patients' MSCs was significantly higher than in donors (11.4 ± 2 x 106 per flask versus 6.9 ± 1.1, p = 0.04). It remained elevated after therapy (10.2 ± 1.5). At the same time, the MSCs population-doubling level per day was significantly decreased in patients in comparison with donors (0.6 ± 0.03 vs. 0.4 ± 0.04, p MFI analysis of antigens typical for MSCs revealed significant differences in the patients MSCs. CD54 (ICAM1) was 1.5 times higher in the onset of the disease compared to donors. It increased 6.3 times (456 ± 124 versus 72 ± 16, p Conclusion Despite the absence of proven BM damage in DLBCL patients, the properties of MSCs - niche components in the stromal microenvironment that regulates hematopoiesis, had been changed. The materials were supported by the Russian Foundation for Basic Research grants, Projects No. 17-00-00170, 17-00-00172. Disclosures No relevant conflicts of interest to declare.

Relapses of Hodgkin's disease (HD) occur in 10-15% of patients with a local tumor and in 20-40% of patients with advanced stages. A relapse of HD is early in half of the patients whereas a second remission can be achieved in only 50% of cases. Autologous hematopoietic stem cell transplantation (auto-SCT) is considered to be the standard of consolidation therapy for relapsing and primary-resistant HD. In patients with relapses of HD, progression-free survival and overall survival are 50% to 60% and 50% to 80%, respectively. Objective. To assess the efficacy of different second-line therapies with subsequent autologous stem cell transplantation in the treatment of relapsing or refractory HD. Materials and methods: from 2000 to 2017, 76 patients with relapsing/refractory HD treated with at least first-line therapy were involved in the study with most of the patients (70%) having received various modifications of BEACOPP regimen. The median age of patients was 27 years. A majority of participants had advanced stages of the disease: 23 (30%), 14 (18%), and 35 (46%) of patients have stage I-II, III, and IV, respectively. Results: DHAP-regimen was given as a second-line treatment in 34 patients resulting in progression, an absence of effect, and overall response in 11 (32%), 13 (38%), and 9 of the patients, respectively. Of them, a complete remission was achieved in only 4 patients, who further underwent autologous stem cell transplantation as a consolidation treatment. Dexa Beam regimen was given as a second-line treatment in 53 patients including those with progression after DHAP-courses. As a result, partial/complete remission, progression, and stable disease were achieved in 35 (66%), 7, and 11 patients, respectively. Auto-SCT was carried out in 32 patients of the overall response group; 28 (88%), 3, and 4 patients have a complete long-term remission, progressive disease, and early relapses, respectively. In the general study population, overall and event-free survival rates were respectively 86% and 71% with a follow-up median of 83 months. Thus, Dexa Beam given as an induction treatment before auto-SCT showed an advantage in the overall response in relapsing/refractory HD. Implementation of auto-SCT as consolidation treatment after complete/partial remission has been achieved allows a long-term positive response to be attained. Disclosures No relevant conflicts of interest to declare.

Introduction. Favorable therapeutic regimen for pregnant women with primary mediastinal B-cell Lymphoma (PMBCL) is determined by tumor mass, somatic status, period of pregnancy and potential risk of teratogenic effects of medications which penetrate through placenta. Aim. To estimate chemotherapy efficacy of PMBCL in case of pregnancy and its toxicity for fetus. Patients and Methods. Since 2004 to 2015 years in National Research Center for Hematology were treated 106 patients with primary mediastinal B-cell lymphoma. In 8 (7,5%) from them disease debuted during 2-3 trimester of pregnancy, one women had two fetuses simultaneously. Before delivery women underwent induction chemotherapy with VACOPB- regimen in 5/8 cases and with R+DAEPOCH in 3/8 cases. Results. After induction treatment had been performed 6/8 women achieved a partial remission, 1/8 had progression, 1/8 - complete remission. High dose chemotherapy was performed in 7/8 women after 1 month following delivery. It was born 9 children (4 boys and 5 girls). Median body weight was 2000 g. (1300 - 3600 g.). Median growth was 47 cm. (40 - 53 cm). In two cases when rituximab had been administered to women before delivery, children were diagnosed with intrauterine pneumonia. One child, born from woman after VACOPB chemotherapy had thrombosis of superior vena cava. All women continue to be in complete remission of PMBCL, children are practically healthy without malformations. The median follow-up of patients was 40 months (15 - 78 months). Conclusions. Pregnancy is not a contradiction for induction chemotherapy of PMBCL in 2-3 trimesters. Rituximab shouldn't be administered before delivery because of a high risk of infectious in fetus. Disclosures No relevant conflicts of interest to declare.

Introduction. Primary mediastinal B-cell lymphoma (PMBCL) amounts from 0,9 to 4% of Non-Hodgkin Lymphomas. Predominantly suffer young women. All relapses in PMBCL occur as early events. Taking into account unfavorable prognosis in case of relapse or progression of PMBCL (2-year overall survival (OS) is about 15% [Kurvilla J., 2008]), it's necessary to achieve a complete remission (CR) in first line treatment. Aim. To compare an efficacy of R-DA-EPOCH followed by R-DHAP and autologous peripheral blood stem cell transplantation (autoSCT) with BEAM conditioning regimen with modified (m) NHL-BFM-90 chemotherapy protocol in pts with partial remission (PR) of PMBCL. Patients and Methods. In 107 pts (34 males, 73 females) diagnosis of PMBCL was established according to WHO criteria. All pts had stage II of disease according to Ann Arbor classification. 30 pts received R-DA-EPOCH, 77 pts - m-NHL-BFM-90 ± R. Median age of pts in R-DA-EPOCH group was 28 years old (from 20 to 67 years old); in m-NHL-BFM-90 ± R-group - 77 years old. Increased LDH concentration was revealed in 24 from 30 (80%) pts in R-DA-EPOCH group and in 75/77 (97%) pts from m-NHL-BFM-90 ± R group. Bulky mediastinal disease was in 30/30 (100%) pts in R-DA-EPOCH group and in 59/77 (76%) - m-NHL-BFM-90 ± R group. In case of CR after 6 courses according to computer tomography and PET scanning treatment was completed. In case of PR pts underwent 2 R-DHAP courses with autoSCT (BEAM conditioning regimen). All pts from m-NHL-BFM-90 ± R group independently from PET-results stopped treatment. All cases of treatment failure occurred till one year of observation. Kaplan-Meier method was used to calculate OS and relapse-free survival (RFS). OS was defined as time from diagnosis to death from any reason; RFS was defined as time from diagnosis until relapse or death from any cause. Results: After 6 R-DA-EPOCH therapy CR was achieved in 24/30 (80%) pts, 6/30 (20%) pts had PR and underwent 2 R-DHAP courses followed by auto-SCT (BEAM). 2- RFS and OS was 100%. Median follow up was 16 months. In m-NHL-BFM-90 ± R group 10-year RFS and OS were 85% and 92%, respectively. Median follow up was 53 months. Conclusion: High dose pulsed m-NHL-BFM-90-chemotherapy as well as R-DA-EPOCH allows getting high efficacy of primary mediastinal B-cell lymphoma therapy. Although future analysis will show more distant results of R-DA-EPOCH chemotherapy. Disclosures No relevant conflicts of interest to declare.

BACKGROUND: Follicular lymphoma (FL) is one of the most frequent types of B-cell lymphomas and accounts for about 22% of all non-Hodgkin lymphomas in Russia. The disease is usually a long-term condition with frequent relapse. At the same time 15-20% of pts have fast-tumor progression. Patients (pts) of this group die within the first 1.5-2 years from the time of diagnosis. High-dose chemotherapy (HDT) followed by ASCT in the presence of nonfavorable characteristics of FL (fast growth of tumor volume, B-symptoms, third citological grade of tumor, large tumor size, absence of anti-tumor response to R-CHOP courses in the first remission) increases the overall survival and progression-free survival of pts. AIM: To estimate HDT with ASCT efficiency among pts with FL in front-line therapy who received treatment in the National Research Center for Hematology during 2000-2013. PATIENTS AND METHODS: The results of ASCT among 12 pts with FL have been analyzed: 8 male and 4 female aged from 31 to 50 with median age 43. Patients in the main group (11/12) were on the IVth stage of tumor growth. In 6/12 cases the tumor size was more than 6 sm. Among 7/12 pts, besides lesions of lymphatic nodes, extra nodal lesions were found: infiltration in psoas (1/12), kidney (1/12), stomach (1/12), lungs (1/12), thyroid (1/12), pancreas (1/12). In 2/12 cases leukaemization was observed. Based on FLIPI criteria all the pts were divided into three groups: in the first risk group - 5/12 pts, in the second - 3/12, in the third - 4/12. B-symptoms were in the majority of cases (9/12). 9/12 pts were diagnosed with I-II cytological grade of FL, among 3/12 - III A/B. Based on the character of tumor growth the dispersion was the following: nodular tumor growth - 5/12, nodular-diffuse tumor growth-6/12, diffuse tumor growth - 1/12. Immuno-chemical investigation of protein serum and urine was performed in 9/12 cases, and among them rising serum β2-microglobulin above the norm was observed in 4/9 pts. A rise in lactate dehydrogenase concentration above the norm was observed among 4/12 pts. Lesion of bone marrow at the beginning of the disease was identified among half of the pts (6/12). Induction courses were performed on the R-CHOP programs, with intensive therapy - on protocol mNHL-BFM-90. RESULTS: Anthracycline courses were prescribed for nearly all of the pts as an inductive therapy: R-CHOP (11/12). mNHL-BFM-90 protocol was chosen as an inductive regime because of the fast growth of tumor size, IIIB grade of FL in 1/12 case. This tumor growth behaved like diffuse large B-cell lymphoma. In three cases medical-diagnostic splenectomy was undertaken before chemotherapy. After the inductive regimes the pts were given HDT based on the following scheme: two courses R-DHAP, course with rituximab and high-doses cyclophosphamide, a course with rituximab and high doses of etoposide, R-BEAM. All 12 pts, who were given ASCT are in full remission on the main disease. The period of observation is from 13 to 164 months. CONCLUSION: This first experience of intensive therapy in FL in Russia demonstrates that ASCT as a front-line therapy leads to complete remission of FL among pts who have nonfavorable prognosis factors. Disclosures No relevant conflicts of interest to declare.

Introduction: Detection of bone marrow involvement (BM) is a factor of poor prognosis for diffuse large B-cell lymphoma (DLBCL), since DLBCL BM is characterized by aggressive course and low response level to standard chemotherapy (CT), and 5-year overall survival (OS) rate for this group is less than 30% after ÑHOP-like CT. Intensification of therapy in this group of patients can improve the results, except patients with bone marrow involvement at diagnosis. Bone marrow involvement is a poor prognostic factor for patients treated with high-dose chemotherapy. Bone marrow involvement in DLBCL by histology is detected in 10-30% patients. The importance of B-cell clonality examination in bone marrow as prognostic and staging factor has not been described in the literature. Aim: To evaluate the significance of the immunoglobulin heavy chain gene rearrangement analysis performed by PCR for identification of the bone marrow involvement frequency and its value for staging and prognosis in patients with de novo DLBCL treated with high-dose chemotherapy (HDC). Patients and methods: We performed a pilot prospective trial, including 175 consecutive adult patients (median age 45 years, range 18–67) with newly diagnosed DLBCL who were enrolled in HDC protocol (mNHL-BFM-90 program or scheme R-EPOCH/R-HMA) since June 2007 till July 2014. Their clinical characteristics included such factors as IPI and phenotype DLBCL by immunohistochemical study. Out of the 175 patients, 85 had a GCB phenotype and 90 - non-GCB; 36 patients (20%) had low IPI risk, 40 patients (23%) had low-intermediate, 38 patients (22%) had high-intermediate, 61 patients (35%) had high risk. B-cell clonality was evaluated using PCR amplification by IGH (FR1, FR2, FR3) and IGK (Vκ-Jκ, Vκ/intron-Kde) gene rearrangements with multiplex BIOMED-2 primer sets and subsequent fragment analysis using ABI PRISM 3130 Genetic Analyzer (Applied Biosystems).In 105 of 175 patients the study by BIOMED-2 multiplex polymerase chain reaction protocol of B-cell clonality of bone marrow was fulfilled. Statistical analysis was done using JMP ver. 10.0 (SAS, Cary, NC). Results: Histological and immunohistochemical studies validated bone marrow involvement in 19 patients (10.8%), including: concordant in - 12 patients (63%), and discordant - in 7 patients (37%). In 14 of these patients the B-cell clonality detection was performed and confirmed immunoglobulin (IG) heavy chain gene rearrangement in all cases. In 26 out of 105 patients, bone marrow involvement (24.7%) was revealed: in 14 patients bone marrow involvement was identified by histological examination and in 12 patients only by clonality of bone marrow. 12 patients with only B-cell clonality in bone marrow were classified by IPI: 4 (33%) patients had high IPI, 6 (50%) patients - high-intermediate IPI and low-intermediate IPI - 2 (17%) patients. So, in 17% patients had to change the risk group according to bone marrow involvement. Thus, 31 patients with bone marrow involvement signs were identified. Univariate analysis of the entire cohort (n = 175) revealed that both IPI and phenotype were not of prognostic significance (p > 0.05). In multivariate analysis, detection of B-cell clonality in bone marrow was an independent predictor of OS and relapse-free survival (RFS) (p < 0.005). (Fig.1-2). OS, RFS in patients with histological detection bone marrow involvement and with only B-cell clonality in the bone marrow did not differ. Conclusion:Detection of B-cell clonality in the bone marrow seems to be an independent predictor of outcome in de novo DLBCL pts, treated with high-dose chemotherapy, while IPI and phenotype DLBCL cannot be considered as risk factors for this group of pts. It seems to be reasonable to include the detection of B-cell clonality in the bone marrow in primary diagnostics of the DLBCL for staging and predicting response in HDC. Figure 1 Figure 1. Figure 2 Figure 2. Disclosures No relevant conflicts of interest to declare.

Abstract 2034 Background: Mantle cell lymphoma (MCL) is aggressive B-cell neoplasm diagnosed predominantly among elderly men. (R)CHOP-like schemes are effective in remission induction, but the progression-free survival is disappointingly short (median 16–20 months) with median overall survival of 3–4 years. Upfront use of high-dose cytarabine (12 g/m2), autoSCT and rituximab at all stages of therapy is the most effective treatment but possible only with patients younger 65 years. Decrease in AraC doses to 4 g/m2 per cycle significantly reduce progression free survival. Prominent efficacy of gemcitabine-oxaliplatin combinations and irinotecan in relapsed and refractory MCL patients allowed including these drugs in first-line treatment in cases when the scheme R-HD-Met-AraC (Romanguera J. 2005) is impossible. Aim: Toxicity and efficacy assessment of schemes R-DA-EPOCH/R-GIDIOX and R-DA-EPOCH/ R-HD-Met-AraC in primary MCL patients eligible for autoSCT. Patients and Methods: Since May 2008 35 untreated MCL patients (median 55 years (29–63), males/females 68,5%/31,5%, MIPIb: 34% low, 26% intermediate, 40% high risk) were enrolled. After first R-EPOCH course (Wilson W. 2003) patients were stratified according to toxicity they had received either R-DA-EPOCH/R-HD-Met-AraC or R-DA-EPOCH/R-GIDIOX. In the absence of hematological toxicity grade 4 for more than 3 days, severe infection complications and signs of renal failure patients underwent treatment under the scheme R-HD-Met-AraC (rituximab 375 mg/m2 day 0, methotrexate 1000 mg/m2 24 hours CI day 1, cytarabine 3000 mg/m2 q 12 hrs days 2–3). If there was one of these complications patients underwent treatment under the scheme R-GIDIOX (rituximab 375 mg/m2 day 0, gemcitabine 800 mg/m2 days 1 and 4, oxaliplatin 120 mg/m2 day 2, irinotecan 100 mg/m2 day 3, dexamethasone 10 mg/m2 IV days 1–5, ifosfamide 1000 mg/m2 days 1–5). Further these courses are rotated: either R-DA-EPOCH/R-HD-Met-AraC or R-DA-EPOCH/R-GIDIOX. Depending on the terms of response, patients received 6–8 courses (3–4 cycles) of chemotherapy and autoSCT (BEAM-R) with in vivo purging by rituximab before harvest and reinfusion. Patients with residual tumor after autoSCT were consolidated with local radiotherapy. Rituximab maintenance was performed every three months for 3 years. The protocol was approved by the local ethics committee. Patients were analyzed in an intent-to-treat basis. Overall survival (OS) and event-free survival (EFS) rates were estimated (± standard error) by using the Kaplan-Meier method. Efficacy of the therapy was assessed by Cheson's response criteria (2008). Toxicity assessment was performed 93 R-DA-EPOCH, 60 R-HD-Met-AraC and 46 R-GIDIOX courses. Results: A median follow-up is 23 months (range 3–54). Toward August 2012 26 patients underwent autoSCT: 14 from R-HD-Met-AraC arm and 12 from R-GIDIOX arm. 1 induction death after first HD-Met-AraC course (acute renal failure and septic shock). Maintenance therapy with rituximab was completed in three patients. All patients achieved CR in R-HD-Met-AraC arm. In R-GIDIOX arm OR was 100%: 11 CR and 1 PR (without progression for 26 months after autoSCT). Main non-hematological toxicity of R-GIDIOX was hepatic, with elevated aminotransferases grades 1–2 and 3–4 in 59,5% and 7,1% of courses respectively, without clinical signs. The sources of stem cells were PB in 23 patients and BM in 3 cases of harvest failure after R-GIDIOX. Hematological toxicity of R-GIDIOX course: leukopenia grade 4 was in 71,4% (medium duration was 5,4 days, range 1–13), thrombocytopenia grade 4 was in 42,9%. The estimated 4-years OS rates for the R-GIDIOX group and the R-HD-Met-AraC group were 100% and 68% ± 17%. The estimated 4-years EFS rates for the R-GIDIOX group and the R-HD-Met-AraC group were 90% ± 10% and 69% ± 14%. Conclusions: Our main goal was to incorporate intensive induction, autoSCT and rituximab maintenance in first line therapy MCL patients. However, HD-Met-AraC scheme is highly toxic and its use is possible only in 2/3 of patients younger 65 years. R-GIDIOX scheme is less toxic than R-HD-Met-AraC and equally effective in response induction and mobilizing, so it could be recommended for those in whom high-dose AraC and methotrexate can potentially cause severe adverse consequences. Such an integrated approach might lead to a shift of paradigm of MCL from an incurable to a curable lymphoma. Disclosures: No relevant conflicts of interest to declare.

Abstract 2684 Introduction: diffuse Large B-cell lymphoma (DLBCL) are aggressive but potentially curable with multi-agent chemotherapy. The R-CHOP regimen is standard therapy for treatment of DLBCL patients. Whereas attempts to improve outcome with more intensive chemotherapy failed to show additional benefit. Aims: to set the indications to standard or intensive therapy in patients with nodal DLBCL and to evaluate the efficiency of modified protocol NHL-BFM-90 in adult patients. Patients and methods: 139 patients with newly diagnosed nodal DLBCL were included in the study since January 2002 till December 2010. The diagnosis was established according to the criteria of the WHO classification. Diagnostic methods include neck, thorax, abdomen and pelvis CT scan, tumor biopsy bilateral bone marrow (BM) biopsy, lumbal puncture, MRT or scintigraphy of bones. Bone marrow involvement was found in 18 cases and confirmed by immunohistochemistry. Bulky disease, direct invasion to adjacent organs, III-IV stage according Ann-Arbor and increased level LDH were considered as the unfavorable prognostic factors. The patients were stratified into 2 groups: the first group with unfavorable prognostic factors (86 patients), the second group without unfavorable prognostic factors (53 patients). Median age in the first group was 53 years. There were 46 men and 40 women. The level of LDH was elevated in the 61 patients. III-IV stage without involvement of BM was diagnosed in the 35 patients, IV stage with BM involvement-in the 18 patients, bulky disease and direct invasion to adjacent organs-in the 33 patients. Median age in the second group was 55 years. There were 28 men and 25 women. All patients in the first group received intensive therapy by modified protocol NHL-BFM-90. The patients in the second group received chemotherapy CHOP-21(n=28) or R-CHOP-21 (n=25). The program NHL-BFM-90 was modified: the dose of methotrexate was 1,5 g/m2 for 12 hours and adriamycin 25 mg/m2 was infused in 1–2 days of course AA. All patients treated with this protocol gave written informed consent. We made spleneectomy to all the patients of splenic DLBCL before chemotherapy (n=20). The estimation of the chemotherapy efficiency was carried out according to the criteria of the International Working Group: rate of complete remissions (CR), overall (OS) and relapse-free (RFS) survival. The data were analyzed with SAS statistical package program. Statistical significance of the difference in the survival curves for two groups was calculated with Log-rank Test. Results: 64 patients in the first group (74,4%) and 53 patients (100%) in the second group achieved CR. 5-year OS and RFS were 65% and 86% in the first group 96% and 100% in the second group, accordingly. In the first group the best results were obtained in patients without BM involvement. 5-year OS was 84% in these patients comparing 12% in patients with BM involvement–figure 1 (Log-Rank Test=0,0001). The rate of CR and OS were the same in CHOP-21 group and R-CHOP-21 group–figure 2 (Log-Rank Test=0,8). Conclusion: The stated unfavorable prognostic factors allowed to stratify patients into prognostic groups. Modified program NHL-BFM-90 is effective treatment for adult patients with DLBCL with unfavorable prognostic factors. The concordant of BM involvement is the most strict prognostic factor. Disclosures: No relevant conflicts of interest to declare.

Abstract 2654 Background. Primary mediastinal (thymic) B-cell lymphoma (PMBL) has been recognized as a subtype of diffuse large B-cell lymphoma (DLBCL). It has its distinctive clinical and morphological features but differs from other types of DLBCL due to peculiar immunophenotype and gene expression profile. PMBL have a distinct gene signature with several highly expressed genes including MAL, JAK2, PDL1, PDL2 and TRAF1. PMBL molecular signature was associated with a more favorable survival compared to DLBCL cases with a non-germinal and germinal-center profile, supporting a distinct natural history for PMBL [A. Rosenwald et al., 2003, G. Lenz et al., 2008]. Aim. The aim of our study was to determine whether the quantitative expression analysis of JAK2, MAL, PDL1, PDL2 and TRAF1 genes may distinguish PMBL cases from other variants of DLBCL with primary involved of mediastinal lymph nodes. Patients and methods. 31 patients with DLBCL with primary involvement of mediastinal lymph nodes and 12 patients with DLBCL without involvement of mediastinal lymph nodes were enrolled in the study. The diagnosis was made by morphological and immunophenotypic analysis of lymph nodes biopsy samples. Six healthy donors of lymphocytes constituted control group. The expression of JAK2, MAL, PDL1, PDL2 and TRAF1 genes was analyzed with RQ-PCR TaqMan hydrolyzing probes. Results. Normal median value of JAK2, MAL, PDL1, PDL2 and TRAF1 gene expression have been established according the data obtained in the set of normal donors. The gene was considered to be overexpressed if its value was more than normal median value + 3SD. In 12 patients with DLBCL without involvement of mediastinal lymph nodes there was no overexpression of MAL, PDL1 and PDL2 genes. In this group only 1 of 12 (8%) pts has the overexpression of JAK2 and 2 out of 12 (16%) pts the overexpression of TRAF1. In patients with DLBCL with primary involvement of mediastinal lymph nodes JAK2 gene was overexpressed in the 18/31 cases (58%), MAL–in the 6/31 (19%), PDL1–in the 2/31 (6%), PDL2–in the 5/31 (16%) and TRAF1–in the 2/31 (6%). Based on the overexpression of more than 3 genes we have separated from DLBCL group (31 cases) a distinct group with primary mediastinal (thymic) B-cell lymphoma 18/31 (58%) patients. So, in PMBL we revealed significant overexpression of at least 3 genes in 18/18 pts whereas in DLBCL with primary involvement of mediastinal lymph nodes no cases have such expression. Summary. Our data suggest that the quantitative molecular analysis of JAK2, MAL, PDL1, PDL2 and TRAF gene expression enables to distinguish PMBL from DLBCL with primary involvement of mediastinal lymph nodes. Disclosures: No relevant conflicts of interest to declare.

To evaluate the efficiency and toxicity of the intensive Burkitt's lymphoma (BL) therapy protocol BL-M-04.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.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.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.Лимфома Беркитта (ЛБ) является одной из наиболее агрессивных В-клеточных лимфом и характеризуется преимущественно экстранодальной локализацией опухоли, частым вовлечением в опухолевый процесс костного мозга (КМ) и центральной нервной системы (ЦНС). Отличительной чертой ЛБ является высокая пролиферативная активность опухолевых клеток (Ki-67 ~100%) и наличие перестройки гена c-MYC в область тяжелых или легких цепей иммуноглобулина. Цель исследования. Оценка эффективности и токсичности интенсивного протокола терапии лимфомы Беркитта ЛБ-М-04. Материалы и методы. С 2003 по 2014 г. наблюдали 70 больных с диагнозом ЛБ: 45 мужчин и 25 женщин в возрасте от 15 до 62 лет, медиана возраста 31 год. I стадия заболевания по S. Murphy диагностирована у 4 (5,7%) больных, II - у 9 (12,9%), III - у 25 (35,7%), IV - у 11 (15,7%), лейкоз Беркитта - у 21 (30%). У 23 (32,9%) пациентов отмечено вовлечение в опухолевый процесс костного мозга, у 15 (21,4%) больных - ЦНС. В-симптомы выявлялись у 56 (80%) пациентов, повышение активности лактатдегидрогеназы (ЛДГ) - у 50 (78,1%) из 64, при этом у 34 (56,2%) из 64 отмечено повышение активности ЛДГ более чем в 2 раза по сравнению с референсными значениями. Медиана активности ЛДГ составила 2398 (238-20 300) ед/л. Острая почечная недостаточность в дебюте заболевания определялась у 17 (24,2%) больных, у 8 химиотерапия начата на фоне терапии, замещающей функцию почек. Лечение проводили по протоколу ЛБ-М-04±R, (4 последовательных блока А-С-А-С±R). С 2011 г. больным с поражением КМ проводили 6 блоков А-С-А-С-А-С с ритуксимабом. Результаты. Полная ремиссия заболевания достигнута у 62 (89%) больных. При этом 6 пациентов умерли от осложнений терапии в период индукции ремиссии, у 2 наблюдалась прогрессия заболевания, у 3 развился рецидив заболевания (у 2 ранний, у 1 через 2 года после завершения лечения). 5-Летняя общая выживаемость (ОВ) составила 85%, 5-летняя безрецидивная выживаемость (БРВ) - 95%. При проведении многофакторного регрессионного анализа Кокса выявлено, что лейкоз Беркитта и поражение КМ являются независимыми факторами, влияющими на ОВ и БРВ. Плохой соматический статус (оценка по шкале ECOG 3-4 балла по сравнению с 0-2 баллами) оказался статистически значимым для ОВ, но не для БРВ. Заключение. Несмотря на оптимистичные результаты, полученные нашей исследовательской группой, необходимы дальнейшее совершенствование протоколов лечения ЛБ, разработка новых подходов к терапии, особенно у пациентов старшей возрастной группы, и больных лейкозом Беркитта.

To evaluate the late cardiotoxicity (CT) of high-dose chemotherapy (CT) according to the modified NHL-BFM-90 (mNHL-BFM-90) protocol in adult patients with diffuse large B-cell lymphoma (DLBCL).The results of electrocardiography (ECG) and echocardiography (echoCG) were analyzed in 40 DLBCL patients treated according to the mNHL-BFM-90 program in the Hematology Research Center (HRC), Russian Academy of Medical Sciences (RAMS), in 2002 to 2009. A study group consisted of 20 men and 20 women whose age was 31 to 76 years; median age was 56.5 years at the time of their examination and the median follow-up time after therapy was 6 years. The individual cumulative dose of doxorubicin was 150-300 mg/M2. A comparison group included 19 patients receiving CHOP/R-CHOP CT in HRC, RAMS, in 2002 to 2009. Out of them, there were 8 men and 11 women whose age was 39 to 78 years median age was 70 years at the time of their examination. The individual cumulative dose of doxorubicin was 200-400 mg/M2. ECG and echoCG were carried out before and 5 years or more after CT.Out of the 40 patients with DLBCL, the signs of subclinical cardiomyopathy (CMP) were detected in 24 (60%) patients; no clinical manifestations of congestive heart failure (CHF) were found in any patient. In the comparison group of 19 patients receiving CHOP/R-CHOP CT, 14 (74%) patients were found to have signs of subclinical CMP and no clinical signs of CHF. The summary toxicity index significantly depended on age (p=0.03) and a history of heart disease (p=0.3); it was significantly higher after CHOP/R-CHOP CT (p=0.05). There was a statistically significant relationship of the risk of subclinical CMP to the history of heart diseases (p=0.05).Late cardiotoxicity of the mNHL-BFM-90 program does not exceed the toxicity of standard CHOP/R-CHOP therapy. Post-CT Echo-CG and ECG findings showed that the patients with the most marked subclinical signs of CMP in both groups had cardiotoxicity risk factors, such as coronary heart disease, hypertensive disease, or diabetes in their history. No clinically significant CHF was identified in any patient.Цель исследования. Оценка отдаленной кардиотоксичности (КТ) высокодозной химиотерапии по модифицированной программе NHL-BFM-90 (mNHL-BFM-90) у взрослых больных диффузной В-крупноклеточной лимфомой (ДВККЛ). Материалы и методы. Проанализированы результаты электро- и эхокардиографии (ЭхоКГ) у 40 больных ДBККЛ, получивших терапию по программе mNHL-BFM-90 в ГНЦ РАМН в период с 2002 по 2009 г. Основную группу составили 20 мужчин, 20 женщин в возрасте от 31 года до 76 лет, медиана возраста 56,5 года на момент обследования, медиана срока наблюдения после окончания терапии - 6 лет. Индивидуальная кумулятивная доза доксорубицина достигала 150-300 мг/м2. Группу сравнения составили 19 пациентов, получивших химиотерапию (ХТ) по программе СНОР/R-СНОР в ГНЦ РАМН в период с 2002 по 2009 г. Из них 8 мужчин и 11 женщин в возрасте от 39 до 78 лет, медиана 70 лет на момент обследования. Индивидуальная кумулятивная доза доксорубицина - 200-400 мг/м2. Электрокардиографию и ЭхоКГ проводили перед началом и через 5 лет и более после окончания ХТ. Результаты. Из 40 больных ДBККЛ признаки субклинической кардиомиопатии (КМП) обнаружены у 24 (60%), клинических проявлений застойной сердечной недостаточности (ЗСН) ни у одного пациента не выявлено. В группе сравнения из 19 пациентов, получивших ХТ по программе СНОР/R-СНОР, признаки субклинической КМП выявлены у 14 (74%) без клинических признаков ЗСН. Суммарный показатель токсичности достоверно зависел от возраста (p=0,03), наличия заболеваний сердца в анамнезе (р=0,03), и был достоверно выше (р=0,05) после ХТ СНОР/R-СНОР. Выявлена статистически значимая (р=0,05) зависимость риска развития субклинической КМП от наличия заболеваний сердца в анамнезе. Заключение. Отдаленная КТ программы mNHL-BFM-90 не превышает токсичность стандартной терапии СНОР/R-СНОР. У пациентов с наиболее выраженными субклиническими признаками КМП после ХТ по данным ЭхоКГ и электрокардиографии в обеих группах имелись факторы риска КТ - ишемическая болезнь сердца, гипертоническая болезнь, сахарный диабет в анамнезе. Клинически значимой застойной сердечной недостаточности ни у одного пациента не выявлено.

Background. TP53 mutations were described to have a negative impact on prognosis of patients with B-chronic lymphoid leukemia and diffuse large cell lymphoma. However, a role of TP53 mutations in high-grade B-cell lymphomas (HGL) is not well defined in context of other genetic aberrations. Materials and methods. 23 patients (7 males and 16 females) had diagnosis of HGL were treated in National Research Center for hematology, Moscow, Russia. Median age was 49 years old (30-76). 5 patients had HGL with c-MYC (MYC-R) and BCL2 genes rearrangements and 2 - HGB with MYC-R and BCL6 rearrangements. 17 (74%) patients had MYC-R, 11/23 (48%) had double expresser lymphoma (MYC≥40%, BCL2≥50%) (DE). Median of observation time was 29,1 months (6,3-99,8). 19 (82%) of patients had IPI score 3-5 points. 16 patients underwent LM-B-04 with rituximab (Table 1), 5 - R-(DA)-EPOCH, 2 - R-CHOP-21. In 5 cases autologous stem cell transplantation was performed. Sanger sequencing was performed to identify mutations in exons 5-8 of TP53 gene using DNA extracted from formalin fixed paraffin embedded tissue («Extra-DNA» kit, «Genetechnology» LLC). Primers to TP53 gene were synthesized based on nucleotide sequences data available online on website ncbi.com by «Evrogene». To evaluate an influence of such factors as TP53 mutation (TP53mut), MYC-R, DHL, DE, gender, therapy on overall survival (OS) and time to progression (TTP) were performed multivariate dispersion analysis and Cox regression analysis (STATISTICA 10). Results: 8 (35%) cases with TP53mut were identified: c.535C>T 45,6% p.H179Y, c.524G>C 15,6% p.R175P, c.743G>A 75,6% p.R247Q, c.487T>A 25,2% p.Y163N, c.824G>A 75% p.C275Y, c.713G>A 87,7% p.C238Y, c.745A>G 31,9% p.R249G, c.639A>G 41,8% p.R213R. 7/8 of them harbored MYC-R, 2/8 had DHL. In univariate (Picture 1, 2) and multivariate analysis pts harboring TP53mut had worse OS (median OS was 6,2 (0,7-9,5) vs 25,5 (0,7-99,8) months, p=0,004) and shorter TTP (median TTP 3,5 (0,7-9,5) vs 23,1 (0,7-99,8) months, p=0,027) than patients without TP53mut. DHL status had also an adverse effect on OS with lower significance than in pts with TP53mut (p=0,022). Adverse effect of TP53mut trends to play a role in combination with c-MYC gene rearrangement, thus5 pts TP53mut/MYC-R had shorter TTP than 5 DHL pts (3,4 (1,1-9,5) vs 7,3 (0,6-67,1) months, p=0,07). Conclusion: High-grade lymphoma has a more powerful and independent prognostic factor than double-hit status - TP53 mutation that contribute inferior prognosis. This factor shouldn't be underestimated in routine diagnostics because of its frequency and requirement of a different therapeutic approach. Disclosures No relevant conflicts of interest to declare.

Background: According to current data B-cell lymphoma unclassified (BCLU), intermediate between diffuse large B-cell lymphoma and Burkitt lymphoma represents a highly aggressive type of lymphoma with dramatically bad response for chemotherapy. Cases with translocation of two genes (MYC and BCL2 or BCL6) are divided into double-hit lymphomas (DHL). We need to estimate risk factors to intensify treatment and manage indications for autologous stem cell transplantation (auto-SCT) according to individual characteristics. Aim: To evaluate results of BCLU treatment according to LB-M-04±R protocol in adults younger than 60 years old and CHOP-like regimens in elderly patients (≥60 years old) with auto-SCT in high risk patients group. Patients and Methods: 21 pts observed in National Research Center for Hematology (Moscow) between 2004 and 2014 years were included in a current study. All of them were convenient to BCLU diagnosis criteria according to WHO classification of hematological malignancies (2008). Genetic analysis included: standard karyotyping in 7 pts (6 – lymph nodes samples, 1 sample of cerebrospinal fluid). FISH analysis was performed in 21 pts (in 7 cases on tumor cells; on imprints of tumor in 4 cases, on histologic slides from paraffin blocks - in 10 cases). Taking into account heterogeneity of a common group we divided all pts into 2 subgroups: DHL and non-DHL cases (7 vs. 14 respectively). Pts younger than 60 years old (17 pts) were treated according to LB-M-04±R protocol which included A-C-A-C courses. Course A consisted from dexamethasone 10 mg/m2 i.v. 1-5 ds, methotrexate 1500 mg/m2 12-hours infusion 1st d, ifosfamide 800 mg/m2 1-5 ds, vincristine 1 mg/m2 1st d, doxorubicine 50 mg/m2 3rd d, cytarabine 150 mg/m2 4-5 ds, etoposide 100 mg/m2 4-5 ds; course C included dexamethasone 10 mg/m2 i.v. 1-5 ds, methotrexate 1500 mg/m2 12-hours infusion 1st d, vinblastine 5 mg/m2 1st d, cytarabine 2000 mg/m2 twice a day 2-3 ds, etoposide 150 mg/m2 3-5 ds. Rituximab were indicated before chemotherapy in dosage 375 mg/m2. CNS prophylaxis was made by intrathecal administration of prednisolone 30 mg, cytarabine 30 mg, methotrexate 15 mg in 1st day of each course. When complete remission (CR) was achieved after 2 courses, treatment lasted 4 courses. When tumor regression was diagnosed after 4 courses, treatment continued till 6 courses. 4 pts ˃60 years were managed by CHOP-like regimens ±R. We performed auto-SCT in non-DHL group with signs of poor prognosis (bone marrow involvement, multiple extranodal sites, CR after 6 courses) and in DHL when CR had been achieved after 4 courses. Pts with DHL after auto-SCT received 2 R-EPOCH courses more. As a conditional regimen BEAM was used. An overall survival (OS) as a primary endpoint and event-free survival (EFS) were assessed with using the Kaplan-Meier method (with log-rank test) to estimate an efficacy of treatment. Statistical analysis was performed with SAS 9.3 (SAS Institute Inc. Cary, NC). Results: Studying group included 9 males and 12 females. Comparing pts according DH and non-DH status, DHL group (n=7) consisted of 2 males and 5 females, median age 48 years (30-74), ECOG status was 2.6 (95%CI 1.3-3.9) and non-DH group consisted of 7 male and 7 female, median age was 46 (23-76), ECOG status was 2.4 (95%CI 1.8-3.1). DHL pts had stage II of lymphoma according to Ann-Arbor classification in 1 case, III in 1 case, IV in 5 cases. Non-DHL pts had stage II of lymphoma in 2 cases and IV in 12 cases. Bone marrow involvement was revealed in 2 cases in DHL group and in 5 cases in non-DHL group. More than 1 extranodal site took place in 3 cases of DHL and 8 cases of non-DHL. Survival rates of groups were comparable because they were not significantly different of these characteristics. The 2-year OS and EFS rates were less for DHL pts compared with non-DHL pts: OS: 43 vs. 75%, P=0.24 and EFS: 29 vs. 66%, P=0.09 respectively (Figures 1and 2). Auto-SCT was performed in 2 pts with DHL treated by LB-M-04±R protocol (both pts still be alive) and in 3 pts with non-DHL (1 pt treated by LB-M-04±R and 1 treated by CHOP-like regimen+R are alive in CR and 1 pt treated by LB-M-04±R protocol developed a relapse). Conclusions: Low OS and EFS in BCLU group are caused particularly by DHL cases. We need to enlarge an observation group to confirm benefits of auto-SCT in BCLU pts with signs of poor prognosis. Figure 1 Figure 1. Figure 2 Figure 2. Disclosures No relevant conflicts of interest to declare.