Chronic Lymphoproliferative Diseases Research Articles

The major mechanisms of disruption of the arginine/citrulline cycle in the pathogenesis of cytostatic-induced intestinal injury in patients with chronic lymphoproliferative diseases and methods of their correction

Objective — on the basis of investigation of the role of oxidative stress in disruption of arginine/citrulline cycle with the use of citrulline as a marker of intestinal functional capacity, to provide ground for administration of S‑ademetionine (S‑AMe) in combination with Bifidobacterium infantis 35624 (B. infantis 35624) in patients with chronic lymphoproliferative disorders (CLPD) for prophylaxis of cytostatic‑induced mucositis during chemotherapy (CT). Materials and methods. Examinations involved 38 patients with CLPD, including 8 (21%) females and 30 (79%) males aged 30—76 years. Patients with CLPD and registered diarrheal syndrome against the background of CLPD progression have been enrolled in the study. All patients received CT according to the current guidelines. Depending on the type of concomitant therapy, patients were divided into three groups: group І (n=13) included patients with CLPD, who underwent CT only; patients of group ІI (n=12) received CT and S‑Ame in a dose of 1000 mg/day intravenously for 10 days and then in a dose of 500 mg twice a day for 20 days; patients of group ІII (n=13) during CT received S‑AMe, in a dose of 1000 mg/day intravenously for 10 days, then in a dose of 500 mg twice a day for 20 days with concomitant B. infantis 35624 1 capsule/day for 30 days. Control group consisted of 20 practically healthy subjects (9 (45%) females and 11 (55%) males, aged 22 to 26 years. The examinations were conducted twice: before CT and after the three courses of CT. Blood serum examinations included levels of substances that form a trimethine complex (TBARS) with 2‑thiobarbituric acid, assessment of catalase activity, arginine and citrulline levels and arginase activity. Results. At baseline examinations of patients of groups I, II and III, levels of TBARS in blood serum exceeded the norm in 1.4 (p=0.0479), 1.3 (p=0.0122) and 1.4 times (р=0.026), respectively. Blood serum citrulline level in the group I was in 1.2 times (p=0.0171) lower than in controls. The negative correlation was determined between serum citrulline levels and TBARS levels in the group I (r=–0.52; p=0.02). After the 3rd course of CT in patients of I group, the concentration of TBARS was in 1.3 times (p=0.0002) higher vs baseline values, and in 1.8 times (p=0.0012) higher than in controls. In patients of the II and III groups, serum TBARS levels were in 1.39 (p=0.0005) and 1.46 times (p=0.0002) lower compared to the group I of patients. The application of S‑Ame and B. infantis 35624 combination in group III patients allowed an increase serum citrulline levels in 1.4 times (p=0.0002) compared to group І and in 1.1 times (p=0.001) compared to group ІІ. Conclusions. The conduction of chemotherapy in patients with chronic lymphoproliferative disorders is associated with an increased risk of cytostatic‑induced mucositis, oxidative stress plays a crucial role in its pathogenesis. The application of S‑ademetionine in combination with Bifidobacterium infantis 35624 allows to prevent intestinal injury during chemotherapy.

Immunogenetic Background of Chronic Lymphoproliferative Disorders in Romanian Patients-Case Control Study.

The implications of the genetic component in the initiation and development of chronic lymphoproliferative disorders have been the subject of intense research efforts. Some of the most important genes involved in the occurrence and evolution of these pathologies are the HLA genes. The aim of this study is to analyze, for the first time, possible associations between chronic lymphoproliferative diseases and certain HLA alleles in the Romanian population. This study included 38 patients with chronic lymphoproliferative disorders, diagnosed between 2021 and 2022 at Fundeni Clinical Institute, Bucharest, Romania, and 50 healthy controls. HLA class I and class II genes (HLA-A/B/C, HLA-DQB1/DPB1/DRB1) were investigated by doing high resolution genotyping using sequence specific primers (SSP). Several HLA alleles were strongly associated with chronic lymphoproliferative disorders. The most important finding was that the HLA-C*02:02 (p = 0.002, OR = 1.101), and HLA-C*12:02 (p = 0.002, OR = 1.101) have a predisposing role in the development of chronic lymphoproliferative disorders. Moreover, we identified that HLA-A*11:01 (p = 0.01, OR = 0.16), HLA-B*35:02 (p = 0.037, OR = 0.94), HLA-B*81:01 (p = 0.037, OR = 0.94), HLA-C*07:02 (p = 0.036, OR = 0.34), HLA-DRB1*11:01 (p = 0.021, OR = 0.19), and HLA-DRB1*13:02 (p = 0.037, OR = 0.94), alleles have protective roles. Our study indicates that HLA-C*02:02 and HLA-C*12:02 are positively associated with chronic lymphoproliferative disorders for our Romanian patients while HLA-DRB1*11:01, HLA-DRB1*13:02, and HLA-B*35:02 alleles have a protective role against these diseases.

Immunophenotypic Profile of Chronic Lymphoproliferative Diseases in the State of Rio Grande Do Norte, Northeast, Brazil

INTRODUCTION: Chronic lymphoproliferative diseases (CLPD) are considered as a heterogeneous group of diseases characterized by monoclonal expansion and accumulation of apparently mature lymphocytes, which have a proliferative and/or survival advantage over normal lymphocytes in different organs such as bone marrow, peripheral blood and lymph nodes. This translates into the progressive accumulation of clonal cells and their products, causing peripheral blood and bone marrow lymphocytosis, in addition to lymphadenopathy, splenomegaly or other organomegaly. According to the World Health Organization, which reflects the international consensus and is based on pathological, genetic and clinical factors, lymphoproliferative neoplasms are classified according to the stage of maturation and the lineage where the neoplastic transformation occurs through flow cytometry (FC). OBJECTIVES: The aim of this study was to demonstrate the immunophenotypic diagnostic profiles of patients with CLPD in Rio Grande do Norte, Brazil by FC. METHODOLOGY: Were investigated 884 patients with CLPD of both sexes through CF with a panel of monoclonal antibodies directed to CLPD. In addition, other patient information, such as age, gender, and clinical and hematological data were also collected. RESULTS: The results showed that 89.7% of the cases were B Cell CLPD (B-CLPD) and 10.3% T and natural killer cells (T/NK-CLPD). The distribution of B-CLPD cases were as follows: 487 Chronic lymphocytic leukemia; 18 B-Cell Pro-lymphocytic leukemia; 25 Hairy cell leukemia; 6 variant Hairy cell leukemia; 6 Follicular lymphoma leukemized; 4 Splenic lymphoma with villous lymphocytes; 33 Mantle cell lymphoma; 4 Waldenstrom's macroglobulinemia; 69 Multiple myeloma; 14 Plasma cell leukemia; 11 Burkitt's lymphoma and 117 Leukemic Non-Hodgkin lymphoma (Table 1). The distribution of CLPD-T/NK was as follows: 12 Large granular lymphocytic leukemia; 14 T-Cell prolymphocytic leukemia; 18 Adult T-cell leukemia; 20 Sézary syndrome; and 24 Peripheral T-Cell lymphoma and 2 DLPC-NK (Table 1). DISCUSSION: The B-CLPD are the most commonly observed corresponding to more than 85% of all cases and are characterized by the presence of clonal B lymphocytes with expression of a single type of immunoglobulin light chain (kappa or lambda) in a proportion greater than 1/10, associated the strong positivity of Pam-B antigens such as CD19, CD22 and CD20 and the absence or low expression of Pam-T antigens. However, other antigens such as CD200, CD5, FMC7, CD79b and CD103, among others, may be present or absent, specifically characterizing these entities as Chronic lymphocytic leukemia (weak CD22+; CD200+, CD5+, FMC7-, CD79b-), Hairy cells leukemia (strong CD22+; FMC7+; CD79b+; CD5-; CD200-; CD103+; CD123+; CD11c+ and Follicular lymphoma (strongCD22+/CD20+, CD10+, CD200-, CD79B+, FMC7+). T/NK-CLPD are less frequent and are characterized by the expression of Pan-T antigens CD2, CD3, CD5, CD7, CD28 and TCL-1, with clonality evidenced by strong positivity of TCRα/β in most cases and increase of TCD4+ or TCD8+ lymphocytes in a ratio above 1:10 in most cases. In this group, large granular lymphocyte leukemia (LGLG) is a rare type of T/NK cell CLPD characterized by peripheral and medullary infiltration of large hypergranular lymphocytic cells, comprising about 2-5% of all cases, the most commonly seen being CD8+ T cells. CONCLUSION: The results presented demonstrate that immunophenotyping is a defining technique for clarifying the diagnosis of patients with CPLD, clonality was also successfully characterized in the investigated cases of CPLP of B ,T and NK lineages.

Features of the course and adverse events risk of coronavirus infection COVID-19 in hematological diseases (data from the CHRONOS19 register for the Primorsky Territory, intermediate stage)

To assess the characteristics of the course of coronavirus infection COVID-19 and to determine the risk factors for adverse events in patients of the regional hematological center. As part of an observational prospective cohort study, data from 144 medical records of patients in Primorsky Krai with hematological diseases and COVID-19 were analyzed. The data of the developed standardized questionnaire of the CHRONOS19 study were used. The primary endpoint (adverse outcome) was a composite point that included mortality from any cause during the observation period, development of acute respiratory distress syndrome, hospitalization in the intensive care unit, and the need for mechanical ventilation. A study of the features of the course of COVID-19 in hematological patients showed an increase in the number of adverse events in patients with neoplastic blood diseases, especially in chronic lymphoproliferative diseases and acute myeloid leukemia. Significant predictors of an unfavorable course of COVID-19 include a refractory/recurrent variant of the course of a blood tumor, glucocorticoid therapy as part of the protocol for the treatment of the underlying disease, stage 3-4 lung damage according to computerised tomography scans at the onset of COVID-19, and the presence of diabetes mellitus. Predictors of an unfavorable course of COVID-19 in hematological patients have been identified. Hematological patients in the context of the COVID-19 pandemic require a coordinated interdisciplinary approach involving hematologists and therapists, careful monitoring of clinical and laboratory parameters to reduce the risk of adverse events.

A-159 Role of TCRB1 Antibody in the Evaluation Panel of T Lymphoproliferative Diseases

Abstract Background The diagnosis of T lymphoproliferative diseases is usually a challenge due to the difficulty indifferentiating pathological T cells from reaction cells and also due to the limitation of clonalitytests currently available. TCRB1 is a monoclonal antibody specific for region 1 of the TCR T cellreceptor Beta chain constant, detecting clonal populations of TCRab positive T cells in caseswhere its expression is monotypic. Its application is similar to immunoglobulin light chainrestriction in the detection of B cell lymphoproliferations and in normal T CD3 + TCRab + cells, theexpression of this marker is polytypic. The objective of the present work is to report theexperience of including this antibody in the lymphoproliferative T panel in a private laboratoryin southern Brazil. Methods Samples received with indication of lymphoproliferative disease and lymphocytosis wereevaluated using the tube Lymphocyte Screnning Tube—LST (Euroflow) and were acquired usingFACS CANTO or FACS LYRIC cytometers. Analyzes were performed using Infinicyt software. After, all, samples that showed alteration in the CD4/CD8 ratio, or antigen alteration in the expressionof pan T markers were evaluated for the complete investigation panel according to Euroflowprotocol. This panel had the incorporation of the tube for the evaluation of clonality that hasthe TCRB1 antibody in the PE channel associated with the anchor markers CD45V500, CD3PerCpcy5, CD4V450 and CD8APCH7. The channels that were not used (FITC, APC, Pecy7) areused when necessary, to evaluate antigens that can help select the desired aberrant population,such as CD5, CD2, CD7 antibodies or to assess maturation, like CD45RA and CD27. Data werecollected from the beginning of the use of this antibody from February 2021 until the presentmoment. Results During this period, 2870 samples for immunophenotyping analysis were received, of which 17cases were T chronic lymphoproliferative diseases. Of the 17 cases found, 11 (65%) were caseswith the pathological population TCRab positive and the expression of TCRB1 could beevaluated. In these cases, the expression of the patologic cells was monotypic for the TCRB1antigen. The cases were: 4 large granular lymphocytic (LGL) leukemia, 4 Adult T-cellleukemia/lymphoma /Peripheral T-cell lymphoma, unspecified , 1 Sezary syndrome, 1 T-celllymphoblastic lymphoma in cortical/common thymocyte stage and 1 T-cell prolymphocyticleukemia (T-PLL). Recent studies have demonstrated the high specificity of this test for clonality, as long as it is associated with an expanded and standardized lymphoproliferative panel. Thepresent cases demonstrated good results for peripheral blood samples, bone marrow and lymphnode biopsy. Conclusion T-lymphoproliferative disorders remain a diagnostic challenge, due to the difficulty of a simpletest to assess clonality. The inclusion of the TCRB1 antibody in the lymphoproliferative panelincreased the specificity of the test for diagnosis, being a method of rapid execution and lowcost. Another advantage of using this marker is the possibility of evaluating small cell clonesthat may not be detected in molecular biology techniques due to the sensitivity of the test.

Expert Consensus on the prevention and treatment of indolent B-cell non-Hodgkin lymphoma with novel coronavirus infection

Indolent B-cell non-Hodgkin lymphoma (B-iNHL) is a group of mature B-cell lymphomas that develop slowly. It is characterized by low immune function and could be risky when complicated with novel coronavirus infection (COVID-19). In order to guide the prevention and treatment of B-iNHL combined with COVID-19, China Anti-Cancer Association Hematological malignancies Committee, the Chinese Society of Hematology Medical Association and Chinese Chronic lymphoproliferative Diseases Working Group formed consensus on COVID-19 vaccination, prognosis, treatment and follow-up of B-iNHL patients for clinician reference.

ЯКІСТЬ ЖИТТЯ ПАЦІЄНТІВ З ХРОНІЧНИМИ ЛІМФОПРОЛІФЕРАТИВНИМИ ЗАХВОРЮВАННЯМИ В ЗАЛЕЖНОСТІ ВІД НАЯВНОСТІ РЕСПІРАТОРНИХ СИМПТОМІВ, СУПУТНІХ ЗАХВОРЮВАНЬ ОРГАНІВ ДИХАННЯ ТА ПОКАЗНИКІВ ФУНКЦІЇ ЗОВНІШНЬОГО ДИХАННЯ

Aim: to determine the indicators of the quality of life in patients with chronic lymphoproliferative diseases (CLPD), depending on the existing respiratory comorbidity, symptoms and the state of the pulmonary function (PF). Materials and methods. 90 inpatients with CLPD were included in the study. All subjects underwent a general clinical examination, evaluation of dyspnea severity according to the mMRC scale and the symptoms severity according to COPD assessment test (CAT). Two questionnaires were used to assess patients’ quality of life: SF-36 and EQ-5D-5L. All patients underwent computer spirometry to assess PF. Data were processed using methods of descriptive and analytical statistics. Results and discussion. Patients with respiratory symptoms had worse indicators of quality of life in domains, reflecting the state of physical health, namely “self-care” (“SC”) (p = 0.008) and “pain/discomfort” (“PD”) (p = 0.04). And the patients with pronounced respiratory symptoms (10 or more points according to CAT) had a significantly lower quality of life according to the indicators of “physical component of health” (“PCH”) (p = 0.0001), “general health” (“GH”) (p = 0.001), “pain” (“P”) (p = 0.002), “role limitation due to physical health” (“RLP”) (p=0.00005), “mobility” (“M”) (р = 0.00007), “SC” (р = 0.00001), “general daily activities” (“GDA”) (р = 0.000001), “PD” (р = 0.000001), “psychological component of health” (“PsCH”) (p = 0.0004), “energy/fatigue” (“EF”) (p = 0.003), “emotional well-being” (“EWB”) (p = 0.01), “role limitation due to emotional problems” (“RLE”) (p = 0.0004), “anxiety/depression” (“AD”) (p = 0.000001). Patients with mixed-type PF disorder with a predominance of obstruction had the lowest indicators of quality of life in the “GH”, “PCH”, “EF”, “SC” and “PD” domains. Patients with mixed PF disturbances with a predominance of restriction had the lowest indicators of quality of life in the domains “EF”, “social functioning” (“SF”), “change in health” (“CH”), “M” and “GDA”. Patients with CLPD who had any of the PF disturbances had lower physical health indicators: “RLP” and “PCH” (p = 0.04 in both cases). Using regression analysis, it was found that decreased FEV1/FVC ratio was a predictor of a lower level of the indicator “PCH”. At the same time, among patients with lymphomas, the presence of obstruction was associated with worse indicators of quality of life in “PD”domains (p = 0.03). Among patients with CLL, the presence of PF obstructive disturbances was associated with a lower indicator of “SF” (p = 0.04). Conclusion: among patients with CLPD, the presence of pronounced respiratory symptoms and disturbances of the PF had a significant negative impact on the quality of life. Thus, there is a need to develop treatment to consider these indicators in order to improve the quality of life of patients with CLPD. Key words: oncohematological diseases, asthma, COPD, pneumonia, comorbidity, leukemi

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Abstract. The aim: to study the prevalence of respiratory symptoms and pulmonary function (PF) in patients with various chronic lymphoproliferative diseases (CLPD) in the Dnipro region of Ukraine. Methods. In all patients who were included in this prospective single-center study, complaints and anamnesis data were collected, anthropometric and general physical examinations, mMRC and CAT questionnaires, analysis of the results of high-resolution computed tomography of the chest organs to determine the presence of intrathoracic lymphadenopathy (ITLAP) were performed. In all patients, parameters of PF were determined using computer spirometry, and when signs of bronchial obstruction were detected, a test with a bronchodilator was performed to determine its reversibility. We analyzed the level of FEV1 , FVC and the ratio of FEV1 /FVC, PEF, MEF25, MEF50 and MEF75. Patients with a confirmed diagnosis of bacterial or viral pneumonia (including patients with COVID-19) were excluded from the study. The methods of descriptive and analytical statistics of the software product STATISTICA v.6.1 (Statsoft Inc., USA) (license number AGAR909E415822FA) were used to process the obtained results. Results. The study found that 37.8 % of patients had complaints from the respiratory system, and 42 % of all patients had significant severity of respiratory symptoms according to the CAT questionnaire. At the same time, only 4.4 % of patients, according to the anamnesis, had established diagnoses of respiratory comorbidity. Among the examined patients with CLPD, 54.4 % had PF disorders, and 81.6 % of them had only obstruction or mixed disorders with a predominance of obstruction (52.5 % mild obstruction, 37.5 % moderate obstruction, and 10 % severe obstruction) and 12.5 % of cases had a positive bronchodilation test. The ratio of FEV1 /FVC was found to be significantly lower in patients with multiple myeloma than in patients with chronic lymph leucosis and lymphomas, which may hypothetically be related to the formation of fibrosis of the bronchial wall; however, the proportions of patients with obstructive PF disorders were equivalent in the groups. ITLAP was detected in 42.2 % of patients. The presence of ITLAP in patients with lymphomas was significantly associated with a lower FEV1 , while in patients with chronic lymph leucosis, who had bronchial obstruction in 43.4 % of cases, the presence of ITLAP had a tendency, but was not statistically significantly associated with the presence obstruction or a lower FEV1 level. Conclusions The obtained results indicate the need for a thorough examination of the respiratory organs, including chest computed tomography and spirometry, in all patients with CLPD, even in the absence of known respiratory comorbidity. According to the obtained results, the presence of PF violations of the obstructive type and the presence of ITLAP are among the main factors that affect the severity of respiratory symptoms in CLPD, therefore, they require careful diagnosis and, in the presence of bronchial obstruction, the appointment of appropriate broncholytic therapy, which can potentially improve the quality life of this category of patients. Key words: multiple myeloma, chronic lymphocytic leukemia, lymphoma, pulmonary function, respiratory symptoms.

Application of modified Matutes score containing CD200 in diagnosis of chronic lymphocytic leukemia.

Immunophenotyping technique is a powerful tool for the diagnosis and differential diagnosis of chronic lymphocytic leukemia (CLL) and other B-cell chronic lymphoproliferative diseases (B-CLPD). CD200 is strongly expressed in CLL. This study aims to analyze the clinical value of modified Matutes score (MMS) containing CD200 in the diagnosis of CLL. We retrospectively analyzed 103 B-CLPD patients diagnosed from January 2020 to July 2021, including 64 CLL patients, 11 follicular lymphoma (FL) patients, 14 mantle cell lymphoma (MCL) patients, 6 marginal zone lymphoma (MZL) patients, 1 hairy cell leukemia (HCL) patient, and 7 lymphoplasmic lymphoma/Waldenstrom macroglobulinemia (LPL/WM) patients. The expression of CD markers between the CLL group and the non-CLL group was compared, and the sensitivity, specificity, and clinical consistency of MMS and Royal Marsden Hospital (RMH) immunophenotyping score system were analyzed. There were significant differences in the expressions of CD5, CD23, FMC7, CD22, CD79b, CD200, and sIg between the CLL group and the non-CLL group (χ2 values were 37.42, 54.98, 30.71, 11.67, 55.26, 68.48, and 17.88, respectively, all P<0.01). When the RMH immunophenotyping score≥4, the sensitivity was 79.7%, and the specificity was 100%. When the MMS≥3, the sensitivity was 95.3%, and the specificity was 100%. The Kappa coefficient of RMH immunophenotyping system was 0.677, and the Kappa coefficient of MMS system was 0.860. The MMS system containing CD200 has better sensitivity and same specificity compared with RMH immunophenotyping system, and MMS system may be more useful in the diagnosis of CLL.

Цитогенетическая диагностика хронических лимфопролиферативных заболеваний с применением локусспецифичных флуоресцентных зондов

Цитогенетическое исследование клеток костного мозга или периферической крови у лиц с лимфопролиферативными заболеваниями является обязательным и необходимым. Результаты помогают врачу с ранней диагностикой заболевания и выбором тактики лечения. Современные молекулярно-цитогенетические подходы, основанные на применении локусспецифичных флуоресцентных зондов (метод FISH), дают возможность быстро получить точную информацию о наличии или отсутствии наиболее распространенных мутаций при определенных нозологиях. Целью настоящей работы является анализ собственных данных цитогенетического обследования c применением «коктейля» из 5 локусспецифичных флуоресцентных зондов лиц с лимфопролиферативными заболеваниями, большая часть которых имела диагноз хронический лимфолейкоз. Цитогенетическое исследование было проведено 16 пациентам в период с 2014 г. по 2022 г. в лаборатории радиационной генетики ФГБУН УНПЦ РМ. Использовали коктейль локусспецифичных зондов: ATM (11q22)/TP53 (17p13), DLEU (13q14)/LAMP (13q34) и cen 12 (12p11.1;12q11.1). 75% обследованных лиц были носителями мутаций. Наиболее часто встречались мутации del17p13 (50%), del13q14 (50%). Считаем, что при наличии у пациента мутации в 13q14 необходимо провести дополнительно исследование на del 13q14.2 (RB1). The cytogenetic assay of bone marrow or peripheral blood cells in persons with lymphoproliferative diseases is mandatory and necessary. The results help a doctor with the diagnosis of the disease and the choice of treatment tactics as early as possible after the patient’s visit to the hematologist. Modern molecular cytogenetic approaches based on the use of locus-specific fluorescent probes (FISH method) make it possible to quickly and accurately obtain information about the presence or absence of the most common mutations. The aim of this work is to analyze our cytogenetic data given with a “cocktail” of 5 locus-specific fluorescent probes in individuals with lymphoproliferative diseases, most of whom were diagnosed with CLL. A cytogenetic study was conducted in 16 patients in the period 2014 - 2022 in the Laboratory of Radiation Genetics of the URCRM. A cocktail of locus-specific probes was used: ATM (11q22)/TP53 (17p13) and DLEU (13q14)/LAMP (13q34) and cen 12 (12p11.1;12q11.1). Seventy five percent of the examined individuals were carriers of mutations. The most common mutations were del17p13 (50%), del13q14 (50%). We believe that if a patient has a mutation in 13q14, it is necessary to conduct an additional study on del13q14.2 (RB1).

Geotrichum Capitatum Infections in the Hematologic Setting

Introduction: Geotrichum capitatum is a ubiquitous fungus in nature and can be responsible for invasive fungal infections (IFI) in immunocompromised patients (pts) in the hematologic setting, most often within the context of a diagnosis of acute leukemia. Approximately 85% of cases of G. capitatum IFI have been reported in Europe (with approximately 85% of which in Italy and Spain), and it has been suggested that the mediterranean climate could be a contributing factor to its epidemiology; considering the limited geographic distribution of reported cases, the largest published multicentric clinical series comprises just 35 pts. The narrow clinical experience with this IFI thus hinders the development of treatment guidelines, and therefore larger series are needed to address these limitations. Methods: We reviewed all fungal infections in our mediterranean Hematology Department in adult pts (18 years and over) over a 10.5 year period from October 2011 to April 2022, selecting cases with positive fungal identifications of G. capitatum by mycologic methods. The date of "Diagnosis” (Dx) was considered to be the moment of collection of the positive sample. Results: We identified 39 patients with G. capitatum isolates. The average number of cases per year was 3.7 (range: 0-6), with a peak of infections in 2014-2015 temporally coinciding with construction work in the main hospital building, in proximity to the Hematology ward. The mean age at IFI-Dx was 59.0 years (range: 23.2-83.2; median: 64.5) and 61.5% of pts were male. The majority of pts had a Dx of acute leukemia - 61.5% (n=24) myeloid (AML) and 7.7% (n=3) lymphoblastic (ALL) -, with a further 5.1% (n=2) with myelodysplastic syndromes (MDS) with excess blasts; 5.1% (n=2) had Burkitt's lymphoma, 12.8% (n=5) had other chronic lymphoproliferative diseases, 5.1% (n=2) had multiple myeloma (MM), and one pt (2.5%) had aplastic anemia. Two-thirds of pts (66.7%, n=26) presented with fungemia in cultures of peripheral blood (PB) or central venous access (CVA) blood (with 38.5%, n=15, in both); 28.2% (n=11) had positive cultures in either sputum or bronchial aspirate (n=1 in both); 7.7% had urinary isolates and one pt each had positive cultures in a wound exudate and in abscess material. The median OS after IFI-Dx was 17 days (range: 0 days-8.5 years); 31% of pts (n=12) died within a week of Dx, half of whom within the first 3 days, often before fungal identification was established; these pts (none of whom were autopsied) were considered a posteriori to have a G. capitatum IFI as their cause of death. A further 33% of pts (n=13) died from a week to a month after Dx; the direct cause of death in this group of pts could not be established in the absence of autopsy, and both the infection and the underlying hematologic disease were considered equally likely causes. A final 21% of pts (n=8) died between one month and one year after IFI-Dx, and the likeliest cause of death was considered to be progressive disease. Five pts (13%), treated with voriconazole+liposomal amphotericin B, are still alive, with no evidence of recurring IFI, and in complete remission (CR) of the underlying hemato-oncology disease: one with ALL at 8.3 months post-IFI-Dx, one with diffuse large B-cell lymphoma at 11.2 months post-IFI-Dx, one with MM at 4.1 years, and two cases of AML, at 7.2 and 8.5 years post-IFI-Dx, respectively. While the ALL pt had fungemia (in both PB and CVA cultures), the other four surviving pts had no evidence of fungemia and had positive cultures in sputum (n=3) and abscess fluid (n=1). Discussion: We present a very large unicentric clinical series, overtaking the largest published multicentric series reported in the literature. In our cohort, the acute leukemias together with high risk MDS account for three quarters of cases. Approximately one third of patients died of active infection in the acute phase of IFI, while one fifth died of progressive hemato-oncologic disease; in one third of patients, who were not autopsied, the cause of death (IFI vs progressive disease) was not established. Only one-eighth of the cohort currently survives in CR; this subset of pts is relevant for the relative absence of fungemia - present in only 20% of pts, vs 66.7% of the full cohort. Our retrospective series of a rare infection, although underpowered to allow statistical analysis (as has been the case with previously published data) relevantly expands the current knowledge base on G. capitatum IFI in the hematologic patient.

Chronic lymphoproliferative diseases and cardiovascular risk (a literature review)

Heart dysfunction that occurred after using of anticancer drugs and monoclonal antibodies may be a limit factor in treatment of chronic lymphoproliferative diseases (CLPD). Cancer therapy-related cardiovascular toxicity include hypotension, hypertension, arrhythmias, conduction disturbances, pericarditis, thromboembolic events, heart failure, death. The risk of cardiotoxicity may be increased by some factors that include drug exposure, age, history of heart diseases, arterial hypertension, drug combination, previous radiotherapy or chemotherapy. The aim of the work is to assess the impact of anticancer treatment on the occurrence of cardiovascular events in patients with CLPD according to the world scientific literature data. It is important to detect the cardiovascular toxicity before the development of clinical manifestations of damage to the myocardium and blood vessels. The role of markers in identifying the risk group of adverse cardiovascular events remains unclear. Early diagnostics and determination of prognostic factors of cardiovascular toxicity, which develop after anticancer therapy of CLPD, are important and not solved problems. Conclusions. The prognosis for the development of cardiovascular events after antitumor treatment of CLPD remains unfavorable. Clinical monitoring, imaging methods, determination of the biomarker levels (natriuretic peptides, troponins) for cardiotoxicity risk stratification are recommended during antitumor treatment to identify early signs and risk of cardiotoxicity. The use of the latest biomarkers and their combinations may be a way to improve the assessment of the cardiotoxicity risk in CLPD. To date, there is no sufficient evidence on the feasibility of routine determining these biomarkers, which indicates the need to plan new studies.

Combined Immune Defect in B-Cell Lymphoproliferative Disorders Is Associated with Severe Infection and Cancer Progression.

B cell chronic lymphoproliferative diseases (B-CLPD) are associated with secondary antibody deficiency and other innate and adaptive immune defects, whose impact on infectious risk has not been systematically addressed. We performed an immunological analysis of a cohort of 83 B-CLPD patients with recurrent and/or severe infections to ascertain the clinical relevance of the immune deficiency expression. B-cell defects were present in all patients. Patients with combined immune defect had a 3.69-fold higher risk for severe infection (p = 0.001) than those with predominantly antibody defect. Interestingly, by Kaplan–Meier analysis, combined immune defect showed an earlier progression of cancer with a hazard ratio of 3.21, than predominantly antibody defect (p = 0.005). When B-CLPD were classified in low-degree, high-degree, and plasma cell dyscrasias, risk of severe disease and cancer progression significantly diverged in combined immune defect, compared with predominantly antibody defect (p = 0.001). Remarkably, an underlying primary immunodeficiency (PID) was suspected in 12 patients (14%), due to prior history of infections, autoimmune and granulomatous conditions, atypical or variegated course and compatible biological data. This first proposed SID classification might have relevant clinical implications, in terms of predicting severe infections and cancer progression, and might be applied to different B-CLPD entities.

Radiological Findings on Chest Computed Tomography in Patients With the Primary Diagnosed Chronic Lymphoproliferative Diseases.

IntroductionThe presence of concomitant respiratory pathology complicates the process of treatment and recovery of patients with chronic lymphoproliferative diseases (CLDs). Therefore, the diagnosis of lung injury is an important step in the management of such patients.ObjectivesThe aim of this study was to determine the prevalence, nature, extent, and location of changes diagnosed by high-resolution chest computed tomography (CT) in patients with CLDs at the initial examination.MethodsMedical records of inpatients who were hospitalized in 2018-2019 to the City Hematology Center of the Public Non-Profit Enterprise “City Clinical Hospital #4” of Dnipro City Council with a confirmed clinical diagnosis of CLDs were included in the retrospective study. The results of initial high-resolution chest CT were studied and analyzed.ResultsOut of 1,004 hospitalized patients with confirmed CLDs, 119 patients were primarily diagnosed. Among them, 81 patients underwent chest CT examination (68.1%) before the beginning of specific therapy. The average age was 65 (56;68) years, 46 (56.8%) were men. 23 (28.4%) patients were diagnosed with chronic lymphocytic leukemia, 28 (34.6%) patients with multiple myeloma, 24 (29.6%) patients with lymphoma, and six patients (7.4%) had other CLDs. It was found that both central and peripheral lymphadenopathy had about a third of the studied cohort of patients (33.3 and 29.6%, respectively), and these symptoms dominated in patients with chronic lymphocytic leukemia (43.5 and 50%, respectively), lymphoma (50 and 52.2%, respectively), and other CLDs (45.8 and 16.7%, respectively), in contrast to patients with multiple myeloma (7.1 and 0%, respectively). Lesions of the lung parenchyma were found in 45.7% of the studied cohort and were met more often in patients with multiple myeloma (67.9%). However, when comparing the prevalence of their categories, no statistically significant differences were found. Predictable bone-destructive changes were statistically significantly more common in patients with multiple myeloma than in other groups of patients with CLDs (P=0.0003).ConclusionsSigns of pulmonary diseases during initial chest CT were found almost in half of the patients with CLDs. It potentially may affect the frequency of treatment complications in such patients. Lymphadenopathy was the most common finding on chest CT, especially in patients with chronic lymphocytic leukemia and lymphoma. And enlarged intrathoracic lymph nodes possibly could lead to pulmonary functions disorders. Among the types of lung parenchyma lesions, pneumofibrosis and foci of consolidation in the lower lung lobes were the most often diagnosed. Chest CT is informative at the stage of the initial examination of patients with CLDs not only for clinical diagnosis but also for the diagnosis of respiratory comorbidities and prediction of the disease outcome and treatment complications.

Pulmonary comorbidity in chronic lymphoproliferative diseases: realities of the problem in the Dnipro region

The prevalence of the chronic lymphoproliferative diseases is increasing worldwide with increase of the population age. It is known that the presence of comorbidities in such patients plays an important role in predicting treatment outcomes. The aim of the work was to study the prevalence and determine the structure of respiratory symptoms and comorbid pulmonary pathology in patients with chronic lymphoproliferative diseases (CLPD) in the Dnipro region of Ukraine. After analyzing 986 cards of inpatients of the hematology department, whose average age was 65 (56; 69) years, it was determined that 9.0% of patients had at least one chronic respiratory disease, the most common among which were chronic bronchitis, chronic obstructive pulmonary disease, as well as community-acquired pneumonia. Dyspnea and tachypnea are also common among patients with CLPD without established respiratory comorbidity, cardiovascular disease, or anemia. Based on the data obtained, we can recommend a study of the respiratory function and pulse oximetry, as well as a thorough collection of anamnesis of smoking and analysis of the results of chest computed tomography in all patients with CLPD in order to identify the possible cause of shortness of breath and establish the presence of respiratory comorbidity.

Use of Flow Cytometry to Evaluate the Immunophenotypic Profile of Patients with Chronic Lymphocytic Leukemia from Rio Grande Do Norte State, Brazil

BACKGROUND: Chronic lymphocytic leukemia (B-CLL) is a clonal proliferation of mature B lymphocytes characterized by indolent clinical course. This clonality is characterized biologically by low expression of surface immunoglobulin (sIg) and restriction to a single immunoglobulin light chain associated with high expression of CD5 antigen and positivity to B cell antigen lymphocytes such as CD19, CD20 and CD23 and negativity to FMC7. The immunological profile and morphological analysis of lymphoid cells are the main means for the differential diagnosis of B-CLL from other chronic lymphoproliferative diseases. OBJECTIVE: The aim of this study was to evaluate the expression pattern of a variety of membrane antigens in leukemic cells originating from patients with B-CLL. METHODS: Peripheral blood samples from 80 patients with B-CLL were analyzed by multiparametric flow cytometry and routine hematologic exams, using a panel of monoclonal antibodies (MoAb): CD45/CD14, CD3/CD19/CD45, CD4/CD8/CD3, CD20/CD5/CD3, CD3/CD16-56/CD45, CD2/CD7, FMC7/CD23, CD103/CD22/CD20, HLADR/CD38, CD10/CD19, CD1a, CD11b in addition to IgM/gD, kappa and lambda immunoglobulin light chains to detect surface immunoglobulin and clonal restriction for immunoglobulin light chains. The hematological data were obtained using a hematological analyzer and cytomorphological analysis in blood film stained with Leishman's stain.RESULTS: The study sample consisted of 45 men and 35 women, ages ranging from 55 to 84 years (mean 65 years). Complete white blood cell count ranged from 10.0 to 42.0 x 109/L. (mean 50.0 x 109/L) and lymphocyte count was greater than 5.0 x 109/l in all cases. The neoplastic cells displayed B-CLL phenotype (CD5+/CD19+/CD20+/HLADR+/CD23+) in the vast majority of the cases, associated with failure to stain for T cell markers (CD1a, CD2, CD4, CD3, CD7, CD8), CD103, CD14 and FMC7. Leukemic cells of most patients also expressed low intensity of IgM and IgD, with clonal restricted kappa light chain in the majority of cases (59.7%). CONCLUSIONS: This investigation highlights the importance of immunophenotyping for correct diagnosis of chronic lymphoproliferative syndromes, and the MoAb panel used was sufficient for diagnostic confirmation of B-CLL. DisclosuresNo relevant conflicts of interest to declare.

Features of the immune status in chronic hepatitis C in patients with lymphopriliferative diseases

Tumor transformation of immunocompetent cells in patients with chronic lymphoproliferative diseases (CLPD) leads to the formation of secondary immune deficiency, which increases the prevalence of infectious complications. Whether the presence of hepatitis C virus (HCV) and the formation of chronic hepatitis (HCV) further weaken the immunoreactivity of the patient, remains poorly understood. The purpose of this work was to reveal the patterns of HCG formation in patients with CLPD with a detailed analysis of immune disorders associated with the chronicity of viral hepatitis C. Another aim of the study was to disclose the patterns of HCG formation in patients with CLP with a detailed analysis of immune disorders associated with the chronicity of viral hepatitis C. The comparative assessment of immunological and biochemical parameters was carried out in 110 patients with HCV (85 – chronic lymphocytic leukemia, 25 – non- Hodgkin lymphoma), with the presence of hepatitis C virus markers (n=20) and 90 patients negative for HCV. To identify intergroup differences, the method of nonparametric statistics was used: the Mann-Whitney criterion. Results. It was found that the group of HCV-infected patients is represented by younger persons compared to uninfected patients (median age – 44 years vs. 60 years, p=0.004). Analyzing the state of the immune system, we have revealed that regardless of the presence of chronic hepatitis C, all patients had a defect in both cellular and humoral immunity. The degree of immune deficiency did not have a direct dependence on viral liver damage. Moreover, in the presence of HCV infection, there was an increase in the microbicidal activity of neutrophils, as well as an increase in the expression of CD95+ on lymphoid cells. Cytokine levels during HCV contamination fluctuated in different directions. There was an increase in the level of ALT in the group of patients with HCV. Findings. The formation of immune deficiency in patients with CLPD is largely associated with the presence of oncopathology and practically does not depend on the presence of chronic hepatitis C.

Analysis of T-Cell Receptor Beta Chain (Tß) Gene Rearrangements Demonstrates the Monoclonal Nature of T-Cell Chronic Lymphoproliferative Disorders

We investigated the rearrangement patterns of the gene coding for the beta chain of the T cell receptor (T beta) in 11 patients with T-cell derived chronic lymphoproliferative disorders, including T-cell prolymphocytic leukemia (T-PLL) and T-cell chronic lymphocytic leukemia (T-CLL). We found that all five cases of T-PLL, and five of six cases of T-CLL, displayed T beta-gene rearrangements, clearly establishing their monoclonal nature. Clonality could not be determined in one case of T-CLL where the T beta gene was found unrearranged. Our results demonstrate that the majority of cases of both clinically aggressive T-PLL and clinically indolent T-CLL are monoclonal. These results suggest that the analysis of T beta gene rearrangements represents a valid tool for the differential diagnosis and clinical monitoring of T-cell derived chronic lymphoproliferative diseases.

Prognostic Role of C-Reactive Protein, C-Reactive Protein Kinetics and C-Reactive Protein/ Albumin Ratio in Newly Diagnosed B-Cell Chronic Lymphoproliferative Diseases

Introduction: C-reactive protein (CRP), the most commonly used clinical indicator of inflammation, plays an important role in disease diagnosis and efficacy evaluation. Recent studies identified elevated CRP level and CRP kinetics, dynamic change of CRP level throughout treatment, were associated with decreased clinical outcome in some malignancies. Because of increased catabolism and the reduce of hepatic synthesis, albumin is oppositely associated with inflammatory. CRP/ albumin ratio (CAR) is a novel and superior prognostic factor involving inflammatory and nutritional factors in various cancers. However, the prognostic role of CRP, CRP kinetics and CRA in B-cell chronic lymphoproliferative diseases (B-CLPD) were not well characterized yet. Our study focused on the prognostic role of CRP, CRP kinetics and CRA in newly diagnosed B-CLPD. Methods: In total, 243 newly diagnosed B-CLPD patients from January 2012 to December 2019 at the Shandong provincial hospital in China were analyzed for overall survival (OS) and disease-free survival (DFS), depending on CRP, CRP kinetics and CRA. OS and DFS were determined by Kaplan-Meier curves and log-rank test. Cox proportional analysis was performed to examine the prognostic significance of clinicopathological variables in multivariate analyses. Results: The five-year OS of patients with elevated pretreatment CRP level (94.3% vs. 56.7%, p&amp;lt;0.001) (Figure 1a), elevated post-treatment CRP level (81.7% vs. 39.4%, p&amp;lt;0.001) (Figure 1c), continuously elevated CRP level during the whole treatment process (80.0% vs. 47.1%, p&amp;lt;0.001) (Figure 1e) and elevated pretreatment CRA level (93.1% vs. 61.9%, p&amp;lt;0.001) (Figure 1g) were shorter than normal patients, respectively. Compared to normal patients, the five-year DFS of patients with elevated pretreatment CRP level (88.0% vs. 33.9%, p&amp;lt;0.001) (Figure 1b), elevated post-treatment CRP level (56.4% vs.35.7%, p=0.020) (Figure 1d), ever-elevated CRP level (62.9% vs. 35.3%, p&amp;lt;0.001) (Figure 1f), continuously elevated CRP level during the whole treatment process (80.2% vs. 35.3%, p&amp;lt;0.001) (Figure 1f) and elevated pretreatment CRA level (87.3% vs. 42.8%, p&amp;lt;0.001) (Figure 1h) were shorter, respectively. Multivariate analyses identified that elevated pretreatment CRP level (HR: 5.110, p=0.001) (Table 1), elevated post-treatment CRP level (HR: 5.826, p=0.006) (Table 2), continuously elevated CRP level (HR: 6.461, p&amp;lt;0.001) (Table 3) and elevated pretreatment CAR (HR: 3.768, p=0.008) (Table 4) had association with worse OS. Likewise, elevated pretreatment CRP level (HR: 3.767, p=0.001) (Table 1), post-treatment CRP level (HR: 2.384, p=0.043) (Table 2), ever-elevated CRP level (HR: 2.425, p=0.027) (Table 3), continuously elevated CRP level (HR: 4.748, p&amp;lt;0.001) (Table 3) and elevated pretreatment CAR level (HR: 2.824, p=0.007) (Table 4) were in independent significance with worse DFS. Conclusions : We demonstrate that CRP level, CRP kinetics and CAR could be potential prognostic indicators with independent significance in patients with B-CLPD. CRP and CAR make an implementation for prognostic evaluation more easily and effectively in B-CLPD patients. Disclosures No relevant conflicts of interest to declare.

Prospective Multicentric Observational Study of COVID19 in Oncohematological Patients in the Catalonia Region: The Opposite Effect of Steroids on Survival

IntroductionCoronavirus Disease 2019 (COVID19) has shown higher mortality in patients with comorbidities, including cancer. First reports from China, Italy, and US showed mortality comprised between 20% and 40% in this specific population. However, the number of patients in these studies was limited and the percentage of hematological patients was underrepresented. In our study, we present a prospective evaluation of patients affected by solid and liquid tumors who were affected by COVID19 from the same geographical area and time period. Survival outcomes, prognostic risk factors, and effects of oncological treatments received were analyzed.MethodsWAll consecutive oncological patients with age > 18 years old affected by COVID19 (confirmed by pharyngeal PCR test) who were hospitalized in 5 tertiary oncological referral centers in the Catalonia region were included in the study. The recruitment period started from 13/03/2020 to 24/04/2020.Two-hundred thirty-one patients were recruited. Thirty-three percent had hematological malignancies and 67% had solid tumors. The median age was 65 years (range 58-75 years). Female patients were 41%. Twenty-six percent of patients were not considered candidates to intensive care treatments (tracheal intubation) due to their advanced oncological status. Of the hematological cohort, lymphoid malignancies represented 36% of the population, multiple myeloma 20%; acute leukemias 15%; chronic lymphoproliferative diseases 12% and other malignancies 17%. At the time of infection, 11% of patients were receiving steroids (more than 0.5mg/kg/24 hours >15 days), 4% were on active immunosuppressive therapy (calcineurin inhibitors, sirolimus or mycophenolate mofetil) and 5% had grade >3 neutropenia. Most importantly, 61% were on active oncological treatment, 18% were on follow-up, 10% had been diagnosed without having received any treatment.ResultsAt COVID19 diagnosis, 75% of patients had a fraction of inspired oxygen (FiO2) < 24% and 63% had fever. Only 16% had cough,16 % had mucous secretion and 12% had dyspnea. All the other symptoms (myalgia, diarrhea, anosmia, ageusia, fatigue, headache) had an incidence < 10%. Chest X-ray was abnormal in 60% of cases. Regarding in-hospital treatments, 85% received antibiotics, 76% hydroxychloroquine, 12% tocilizumab, 30% corticoids, 70% prophylactic anticoagulation and 55% oxygen support.The median duration of hospitalization was 9.00 days (range 5-16 days).WThe overall mortality rate was 26% with a non-significant difference between hematological (32%) and oncological (23%) patients (p-value = 0.23, figure 1). A significant difference was observed between patients who were candidates to intensive care treatments (21.5%) and not candidates (42.6%)(p-value = 0.0038, figure 1).The following risk factors were associated with decreased survival on multivariate analysis: having the oncological disease in progression (HR 3.33, 95%Confidence Interval (CI)= 1.73-6-41, p<0.001); use of steroids (>0.5mg/kg/24 hours) in the last 15 days (HR 3.76; 95%CI=1.69-8.40, p<0.001); age (continuous variable, HR 1.05, 95%CI= 1.01-1-08, p=0.005); grade 4 neutropenia. When considering COVID19 in-hospital treatments, only steroids were associated with a protective effect on survival (HR 0.51, 95%CI=0.27-0.94, p=0.032) while hydroxychloroquine and tocilizumab had no significant effect.Severe respiratory insufficiency (defined as use of >50% FiO2 as oxygen support) was present or developed in 46% of patients. On multivariate analysis, the following factors were associated with a higher risk of developing severe respiratory insufficiency: use of steroids (>0.5mg/kg/24 hours) in the last 15 days (HR 2.36; 95%CI=1.12-4.97, p=0.023); age (HR 1.05, 95%CI= 1.01-1-08, p=0.009) and dyspnea at diagnosis (HR 3.95; 95%CI=1.77-8.78, p=0.001).ConclusionWCOVID19 is associated to increased mortality in patients affected by solid and liquid tumors. Being a candidate for intensive care treatments could improve survival while having progressive disease, older age and grade >3 neutropenia were considered negative factors. Interestingly, the use of steroids was associated to reduced survival if received within 15 days before COVID19 diagnosis, while it has protective effect when used as part of COVID19 therapy.DisclosuresMussetti:Novartis, Gilead: Honoraria, Research Funding. Sureda Balari:Gilead/Kite: Consultancy, Honoraria; Celgene: Consultancy, Honoraria; BMS: Speakers Bureau; Roche: Honoraria; Sanofi: Consultancy, Honoraria; Merck Sharpe and Dohme: Consultancy, Honoraria, Speakers Bureau; Janssen: Consultancy, Honoraria; Novartis: Consultancy, Honoraria; Celgene/Bristol-Myers Squibb: Consultancy, Honoraria; Takeda: Consultancy, Honoraria, Speakers Bureau; Incyte: Consultancy.OffLabel Disclosure:Tocilizumab for COVID19