Aim. To assess the number of plasma cells (PC) in the bone marrow and their immunophenotype using flow cytometry (FC) and light microscopy. To analyze clinical and prognostic value of the data obtained in newly diagnosed multiple myeloma (MM) patients treated with first-generation proteasome inhibitor bortezomib. Materials & Methods. The study enrolled 153 newly diagnosed MM patients treated and followed-up at the IP Pavlov First Saint Petersburg State Medical University in the period from 2007 to 2017. The median age of patients was 69 years. In 115 patients, the regimens based on first-generation proteasome inhibitor bortezomib were used as induction therapy. To determine the immunophenotypic profile of PC, the CD19, CD20, CD27, CD38, CD45, CD56, CD138, and CD117 monoclonal antibodies were used. PC immunophenotyping in the bone marrow was performed by FC using Cytomics FC500 (Beckman Coulter, USA). Results. Patients with different phenotypes did not show any considerable differences in monocloncal production of certain classes and types of immunoglobulin heavy and/ or light chains. In case of immunophenotypic profile of CD20+CD27- myeloma cells, the secretion of the monoclonal к-chain predominated over that of Л-chain. By and large, the secretion of light chains was observed more often in ММ CD20+ and more seldom in ММ CD56+. In case of CD56 expression, IgM secretion was more often reported; IgAK secretion was more common in case of CD117 expression. Worst survival scores were shown by patients with PC immunophenotype CD27-CD56-. At the primary MM diagnosis, the advanced stages of the disease, according to the ISS, were more commonly characterized by phenotype CD45-CD27-CD56+. Conclusion. The flow cytometry characteristics of PC immunophenotype can be applied to evaluate the prognosis of MM and to optimize the therapy.

Цель. Оценить с помощью методов проточной цитометрии (ПЦ) и световой микроскопии количество плазматических клеток (ПК) в костном мозге и их иммунофенотип. Проанализировать клиническое и прогностическое значение полученных данных у пациентов с впервые диагностированной множественной миеломой (ММ), получавших лечение на основе ингибитора протеасомы первого поколения бортезомиба. Материалы и методы. В исследование включено 153 пациента с впервые диагностированной ММ, проходивших лечение с последующим наблюдением в ПСПбГМУ им. И.П. Павлова в период с 2007 по 2017 г. Медиана возраста пациентов 69 лет. В качестве индукционной терапии у 115 пациентов применялись схемы на основе ингибитора протеасомы первого поколения бортезомиба. Для определения иммунофенотипического профиля ПК использовались моноклональные антитела CD19, CD20, CD27, CD38, CD45, CD56, CD138, CD117. Иммунофенотипирование ПК в костном мозге проводили методом ПЦ на приборе Cytomics FC500 (Beckman Coulter, США). Результаты. Значительных различий в моноклональной продукции отдельных классов и типов тяжелых и/или легких цепей иммуноглобулинов у больных с различным фенотипом не выявлено. При иммунофенотипическом профиле миеломных клеток CD20+CD27–преобладала секреция моноклональной цепи κ над λ. В целом секреция легких цепей чаще отмечалась при ММ CD20+, реже — при ММ CD56+. При экспрессии CD56 чаще наблюдалась секреция IgAλ, а при экспрессии CD117 — IgAκ. Наихудшие показатели выживаемости оказались у пациентов с иммунофенотипом ПК CD27–CD56–. Поздние стадии заболевания по системе ISS на этапе первичной диагностики ММ чаще характеризовались фенотипом CD45–CD27–CD56+. Заключение. Особенности иммунофенотипа ПК, выявленные по результатам ПЦ, могут использоваться у пациентов с ММ для определения прогноза и оптимизации терапии.

Decreased activity of glucocerebrosidase (GCase) as a result of mutations in the GBA gene causes Gaucher’s disease (GD), which belongs to the group of lysosomal storage disorders. The risk of Parkinson’s disease in homo- and heterozygous carriers of GBA mutations is elevated seven- to eightfold. Screening of novel compounds designed to enhance GCase activity requires development of in vitro models based on primary cell cultures obtained from patients carrying GBA mutations. In this work, the efficiency of different methods used to culture peripheral blood macrophages of GD patients and control subjects was compared, and GCase activity and lysosphingolipid concentrations were evaluated using tandem mass spectrometry (HPLC‒MS/MS) in dried cell spots. For the first time, the efficacy of restoring the activity of mutant GCase has been assessed in primary macrophages of GD patients cultured in the presence of pharmacological GCase chaperones isofagomine and ambroxol. Based on these results, a convenient method of in vitro screening of candidate pharmacological agents designed to increase GCase activity can be proposed.

Introduction. The main features of bone marrow blasts cells in Burkitt lymphoma/leukemia (BL) are L3 morphology, mature immunophenotype of blasts with surface IgM expression, and presence of typical MYC gene rearrangements.The aim of the study was to show discrepancy examples in laboratory signs of BL.Patients and methods. 10 patients (8 boys and 2 girls) aged 1 to 18 years were included in the present study. The inclusion criterion was the identification of discrepancies between flow cytometric, morphological and cytogenetic data.Results. In 2 cases there were no rearrangements of the MYC gene. In 2 patients, the L2 morphological variant went against the presence of typical MYC gene rearrangements. In one case, undifferentiated blasts cells were described by morphology together with presence of surface IgM, and atypical genetics. In 8 patients, there was no expression of surface IgM. Of these, patients with absence of cytomorphological data cytometric and genetic data were controversial.Сonclusion. The cases presented in this study and the cases described in the literature demonstrate the importance of an attentive and comprehensive approach in evaluating the results of laboratory tests in the diagnosis of BL.

Introduction. Surface immunoglobulin expression is a main immunophenotypic criteria of mature subtype of B-lineage acute lymphoblastic leukemia. Although the majority of such cases represents Burkitt leukemia/lymphoma, it was shown for several times that membrane IgM could be detected in the absence of other mature lymphomas signs.The aim of the study was to evaluate heterogeneity of childhood acute lymphoblastic leukemia (ALL) with surface IgM expression and to assess correspondence of BIV EGIL ALL subtype with Burkitt lymphoma (BL) bone marrow dissemination.Materials and methods. Immunophenotypic, cytomorfologic and genetic data of 54 BIV-ALL cases were analyzed.Results. Among the studied patients 39 had BL, while others belonged to B-cell precursor ALL (BCP-ALL). All BL patients and none of BCP-ALL patients carried C-MYC rearrangement while in BCP-ALL group in 8 cases and in any BL cases KMT2A rearrangements were found. None of BCP-ALL children had L3 morphology according to FAB classification.Conclusions. B-lineage ALL with surface IgM expression is rather heterogeneous group of cases including typical BL and rare cases of BCP-ALL even with KMT2A-rearrangements. Combination of all available diagnostic technologies will allow precise split of these two different disease and select the appropriate treatment scheme.

Comparison of interpretation of acute lymphoblastic leukemia (ALL) flow cytometric diagnostics data was the aim of the study. Immunophenotyping data obtained from 10 patients with ALL were analysed separately in 26 laboratories from Russian Federation and Kazahstan. Results comparison showed four main type of discordance: B-lineage ALL diagnostics during heavy bone marrow regeneration, great variability of T-ALL interpretation, complexity of ambiguous lineage acute leukemia and, finally, very different report types, unique for each laboratory. All these problems are the serious obstacles for standardization of flow cytometric ALL diagnostics in multicenter setting. Continuation of similar QC rounds following by consecutive discussions with further development of consensus diagnostic algorithm could be the first step for standardization of ALL immunophenotyping in Russian Federation and CIS countries.

Острый мегакариобластный лейкоз представляет собой достаточно редкую форму патологии даже в практике специализированных онкогематологических отделений. На основании анализа данных цитометрии 2363 больных, наблюдавшихся в ФНКЦ ДГОИ им. Д. Рогачева и ПСПбГМУ им. акад. И.П. Павлова с 2007 по 2014 г., диагноз острого мегакариобластного лейкоза установлен 24 пациентам различных возрастных категорий, включая 16 детей (11 мальчиков и 5 девочек) и 8 взрослых (соотношение М:Ж 1:1). Трисомия по хромосоме 21 выявлена только у 4 пациентов раннего возраста (из 13 детей младше 3 лет). В статье описаны особенности пробоподготовки и интерпретации данных многоцветной проточной цитометрии с учетом опыта отечественных и зарубежных коллег.

The acute megakaryoblastic leukemia is a rather rare form of pathology even in practice of specialized oncological hematological departments. The data of cytometry analysis of 2363 patients monitored from 2007 to 2014 carried out in The Dmitrii Rogachev Federal research clinical center of children hematology, oncology and immunology and The academician I.P. Pavlov First St. Petersburg state medical university was used to diagnose acute megakaryoblastic leukemia. The corresponding diagnosis was established in 24 patients of various age categories, including 16 children (11 boys and 5 girls) and 8 adults (ratio male/female 1:1). The trisomy on chromosome 21 was revealed only in 4 children out of 13 of early age (younger than 3 years). The article describes characteristics of samples taking preparation and interpretation of data of multicolor with regard to experience of national and foreign colleges.

The authors present results of the investigation of melatonin receptors expression in lymphocytes in dynamics in 102 patients with acute pancreatitis of mild and severe form and in 50 volunteers. A correlated analysis was made between obtained results of laboratory and instrumental researches and clinical course of acute pancreatitis. The decrease of MT1 receptors expression was noted on 25% in patients with acute pancreatitis. The decline of MT2 receptors expression was observed on 40% of patients with acute severe pancreatitis and in a case of acute mild pancreatitis--on 15.5%, respectively. Values of MT1 and MT2 expression were equal between healthy volunteers. The decline of MT2 expression was a prognostic unfavourable sign. Obtained results of dynamic expression assessment of MT-receptors were presented as MT2/MT1 indices. Given index didn't change during disease, because of this, the index could be used as a prognostic development marker of destructive form of acute pancreatitis at the moment of patient's admission to hospital. Mean values of MT2/MT1 were determined for the purpose of universalization of used method (1.13 +/- 0.09 for mild form and 0.81 +/- 0.09 for severe form of acute pancreatitis, respectively).

Immunophenotyping by flow cytometry is an important component of acute leukemia diagnostics. Revised WHO classification of tumors of hematopoietic and lymphoid tissues incorporates new changes related to acute leukemia subclassification. First part of this article is dedicated to the acute myeloid leukemia and represents a comparison of im-munophenotypic features that characterize the subclasses of acute myeloid leukemia recognized by the revised WHO classification.

Melatonin is a mammalian hormone that has a great variety of effects. At present there is evidence that this hormone considerably reduces the manifestations of many gastrointestinal inflammatory diseases. The biological effects of melatonin are realized through the receptors located on the membranes of different animal cells. Three types of melatonin receptors are now known; of them two (MT1 and MT2) receptors were detected in mammals. Varying clinical forms of gastrointestinal diseases may be pathogenetically caused by the quality and amount of MT1 and MT2 receptors, their ratio, and endogenous melatonin activation. The purpose of this study was to develop a procedure for measuring the human blood cell levels of MT1 and MT2 receptors. For this, specific antibodies to MT1 and MT2 receptors were experimentally obtained; then indirect immunofluorescence was used to determine the content and ratio of blood mononuclear cells having these receptors onto the surface in 23 volunteers. The findings are an initial stage of this study and provide considerable opportunity to study a role of melatonin and its receptors in the pathogenesis of many diseases of the human digestive system.