Nonclassical Monocytes Are Prone to Migrate Into Tumor in Diffuse Large B-Cell Lymphoma

Absolute count of circulating monocytes has been proposed as an independent prognostic factor in diffuse large B-cell lymphoma (DLBCL). However, monocyte nomenclature includes various subsets with pro-, anti-inflammatory, or suppressive functions, and their clinical relevance in DLBCL has been poorly explored. Herein, we broadly assessed circulating monocyte heterogeneity in 91 DLBCL patients. Classical- (cMO, CD14pos CD16neg) and intermediate- (iMO, CD14pos CD16pos) monocytes accumulated in DLBCL peripheral blood and exhibited an inflammatory phenotype. On the opposite, nonclassical monocytes (ncMOSlanpos, CD14low CD16pos Slanneg and ncMOSlanneg, CD14low CD16pos, Slanneg) were decreased in peripheral blood. Tumor-conditioned monocytes presented similarities with ncMO phenotype from DLBCL and were prone to migrate in response to CCL5 and CXCL12, and presented similarities with DLBCL-infiltrated myeloid cells, as defined by mass cytometry. Finally, we demonstrated the adverse value of an accumulation of nonclassical monocytes in 2 independent cohorts of DLBCL.

HUMAN CYTOPENIA VARIANTS AT DIVERSE HIV INFECTION STAGES

Over decades, HIV infection and its complications have been one of the most debated problems in the world. The human immunodeficiency virus leads not only to weakened immune system, but also disrupts normal hematopoiesis manifested as cytopenia (anemia, thrombocytopenia and neutropenia). Materials and methods. A retrospective analysis of cases of combined HIV infection and inhibited hematopoiesis was carried out according to hemogram data of patients admitted for treatment at the IKB No. 1 named after D. Dalmatov, Omsk. The inclusion criteria were cytopenia during hospitalization detected in detailed blood test (by calculating hemoglobin level, counts of erythrocytes, leukocytes, platelets). The age of the patients included in the study differed: from 20 to 29 years - 27 patients (24.6%), from 30 to 39 years - 69 subjects (62.7%), from 40 to 49 years - 13 patients (11.8%), over 50 years old 1 patient (0.9%). All patients had suppression of at least one hematopoietic cell lineage. Anemia was considered as decreased hemoglobin level below than 130 g / l in men and 120 g / l in women. Erythrocytopenia was considered as decreased erythrocyte count below 4.76x10 * 12 / l. Leukopenia was defined as decreased total count of leukocytes below 4.0x10 * 9 / L, while a decrease in the absolute count of neutrophils below 1000 cells / μL was considered as neutropenia. Thrombocytopenia was determined as decreased platelet count below 150x10 * 9 \ l. Results. All patients had suppression of at least one hematopoietic cell lineage. 6 patients with stage 2 had one-cell lineage cytopenias, 7 – two- cell lineages. While analyzing the data obtained, it can be concluded that in patients with stage 2 HIV, inhibition of erythroid and platelet cell lineage predominates, whereas thrombocytopenia reached grade IV. At stage 3 HIV, all 7 patients had inhibition of only one cell lineage. In this group, the inhibition of hematopoiesis had a lighter degree in all hematopoietic cell lineages. In 46 patients with stage 4, there were various oppression of one of the hematopoietic cell lineages, in 44 patients there were two-cell lineage cytopenias. For patients with a more advanced stage of HIV, a decrease in the number of all cellular elements of the blood in the hemogram is characteristic; these disorders are more severe and persistent.

In vivo immunoloprotective potentials of Plantago lanceolata aqueous extract on methotrexate induced albino male mice

This study aimed to evaluate the immunological potential of Plantago lanceolata through determination of (total and absolute count of white blood cell, total count of red blood cell and total count of hemoglobin). The results indicated the ability of plant extract to modulate toxic effect of methotrexate on albino mice by enhancing immunity through all tested parameters All this effect due to the presence of chemical active constituents of plant especially (flavonoid and alkaloids).

Immunological monitoring in cardiac allograft vasculopathy

Background The interaction of immunological determinants and classic cardiovascular risk factors can accelerate the development of cardiac allograft vasculopathy (CAV) with deleterious consequences for the graft function in heart transplantation (HTx). When it comes to immunological risk assessment, inverse CD4/CD8 ratio can be a poor prognostic marker in coronary artery disease, but its influence is unclear in CAV. Aim To evaluate the role of the T-lymphocyte count in peripheral blood as well as CD4/CD8 ratio as a predictive marker for CAV severity in a very long-term follow-up after HTx. Methods We performed a retrospective analysis of patient data collected during routine clinical follow-up visits. These data included innate and adaptive immune cell count in peripheral blood (lymphocyte count, CD3+, CD4+, CD8+ and CD19+ T cells and NK cells). Results The study population consisted of 174 patients with a mean follow-up of 13.1±6.5 years and a mean age at the time of HTx of 45.2±15.0 years. CAV was diagnosed in 71 patients (40.8%), more than half of which underwent interventional procedure or surgical therapy (n=40, 56.3%). A comparison of the cytoimmunological profile of patients with no CAV or mild disease (group 1, n=134) vs. with CAV requiring treatment (group 2, n=40), revealed significantly reduced percentage of CD4+ T cells (46.4±11.4% vs. 41.2±9.6%, p=0.01) and elevated percentage of CD8+ T lymphocytes in group 2 (28.3±14.1% vs. 35.8±13.7%, p=0.003). Thus, the CD4/CD8 ratio was altered in therapy requiring CAV (2.3±2.0 vs. 1.5±1.0, respectively, p<0.001). However, we observed no differences in the absolute count of T-helper cells (CD4+ T cells: 692.2±329.2 vs. 653.8±390.5 cells/μL, p=0.54) and cytotoxic T lymphocytes (CD8+ T cells: 474.7±450.2 vs. 600.0±469.0 cells/μL, p=0.13). Further analysis showed no differences regarding lymphocyte count and absolute count or percentage of CD3+ and CD19+ T cells as well as NK cells. Inverse CD4/CD8 ratio (<1) was associated with greater risk for therapy requiring CAV (OR 2.8, 95% CI 1.3 – 5.9, p=0.009) in a univariate logistic regression analysis. Conclusions Decreased CD4+ T cell count along with increased cytotoxic T lymphocyte count resulting in inverse CD4/CD8 ratio is associated with increased CAV severity in HTx. Given the possible interactions with the immunosuppressive agents and prednisolone, monitoring of the cytomimmunological profile can help identify patients at risk and be useful in establishing therapeutic strategies. FUNDunding Acknowledgement Type of funding sources: None.

Neutrophil Maturation, Reactivity and Granularity Research Parameters to Characterize and Differentiate Convalescent Patients from Active SARS-CoV-2 Infection

Studying the dynamics changes of neutrophils during innate immune response in coronavirus 2019 (COVID-19) can help understand the pathogenesis of this disease. The aim of the study was to assess the usefulness of new neutrophil activation parameters: Immature Granulocyte (IG), Neutrophil Reactivity Intensity (NEUT-RI), Neutrophil Granularity Intensity (NEUT-GI), and data relating to granularity, activity, and neutrophil volume (NE-WX, NE-WY, NE-WZ) available in hematology analyzers to distinguish convalescent patients from patients with active SARS-CoV-2 infection and healthy controls (HC). The study group consisted of 79 patients with a confirmed positive RT-PCR test for SARS-CoV2 infection, 71 convalescent patients, and 20 HC. We observed leukopenia with neutrophilia in patients with active infection compared to convalescents and HC. The IG median absolute count was higher in convalescent patients than in COVID-19 and HC (respectively, 0.08 vs. 0.03 vs. 0.02, p < 0.0001). The value of the NEUT-RI parameter was the highest in HC and the lowest in convalescents (48.3 vs. 43.7, p < 0.0001). We observed the highest proportion of NE-WX, NE-WY, and NE-WZ parameters in HC, without differences between the COVID-19 and convalescent groups. New neutrophil parameters can be useful tools to assess neutrophils’ activity and functionalities in the immune response during infection and recovery from COVID-19 disease.

Abstract 46: Vascular Endothelium-derived Endothelin 1 Promotes Salt-induced Kidney Inflammation In Male But Not Female Mice

Endothelin 1 (ET-1), potent vasoactive and pro-inflammatory peptide, plays a criticalrole in hypertension and end-organ damage. The inflammatory response to high saltintake displays sex differences; however, if a sex difference exists in vascular ET-1inducing salt-induced kidney inflammation is unknown. We aimed to assess the role ofendothelium-derived ET-1 in promoting kidney inflammation in male and female micefed high salt. Aged-matched male and female vascular endothelial cell ET-1 knockout(VEET KO) and control floxed ET-1 (VEET fl/fl ) mice were fed 4.0% NaCl chow (HSD).After 3 weeks of HSD, kidney myeloid and lymphoid cell populations and their functionalstatus were examined by flow cytometry. In response to HSD, male VEET fl/fl mice hadsignificantly higher kidney TH17 cell numbers than female VEET fl/fl mice (IL-17A + CD4 + cell absolute count, male vs. female: 229.6±27.7 vs. 84.9±17.2, p=0.0003, n=6-10/group), indicating a substantial sex-dependent difference in kidney TH17 cellabundance. Lack of endothelium-derived ET-1 resulted in 72% reduction in renal TH17cell number in males (VEET fl/fl vs. VEET KO: 229.6±27.7 vs 64.0±64, p=0.0002), but notin females (p>0.05). Frequency of kidney IL-17A + CD4 + cells was not different betweengenotypes in females or between male and female VEET fl/fl mice. However, male VEETKO mice displayed 57% less renal IL-17A + CD4 + cells than male VEET fl/fl (respectively,0.6±0.071 vs. 1.4±0.2, p=0.0055), and 59% reduction compared to female VEET KOmice (1.4±0.2, p=0.0052). Assessment of the effects of vascular ET-1 on the innateimmune response showed that activated kidney resident macrophages(F4/80 hi CD11b + CD64 + ) were increased in male vs. female VEET fl/fl (MFI, respectively,2080±105 vs. 1672±25, p=0.0002). Lack of endothelium-derived ET-1 significantlyreduced CD64 abundance in males (p=0.012), but not females (p>0.05), whencompared to VEET fl/fl . In conclusion, endothelium-derived ET-1 is critical in promotingrenal innate and adaptive inflammatory responses in males in response to high salt, andimportant male-female differences exist in the manner by which vascular ET-1 regulateskidney inflammation in this setting. Funded by NIH F31 HL151264-0 to PAM,P01HL136267 to JSP, and K01HL145324 to CDM.

Evaluation of the Relationship Between Epidemiological Factors and Laboratory Findings With the Cause of Neutropenia in Children: A Cross-Sectional Study

Background and Objectives: Neutropenia is the absolute count of neutrophils less than 1500 per cubic millimeter. Because the early detection of the cause of neutropenia and appropriate measures to reduce its mortality and financial costs are important, this study was conducted to investigate the relationship between the cause of neutropenia and the severity of neutropenia with clinical and laboratory findings to take appropriate measures. Subjects and Methods This study was a cross-sectional descriptive-analytical study. In this study, 111 patients with neutropenia were studied in Hazrat Masoumeh Hospital in Qom City, Iran, by a census method in 3 years from 2014 to 2016. Necessary information was obtained from the patients' medical records through a questionnaire. Data were analyzed by SPSS software. Results The study findings showed relationships between the cause of the disease and variables of age (P= 0.007), the severity of neutropenia (P<0.001), disease outcome (P<0.001), length of hospital stay (P<0.001), Hb (P<0.001), and WBC (P<0.001). The causes of neutropenia in the studied patients were viral (54.1%), sepsis (24.3%), malignancy (10.8%), anemia (4.5%), idiopathic (3.6%) and ITP (2.7%). Conclusion The present study showed a significant relationship between demographic and laboratory findings with the cause and severity of the disease. Therefore, considering these factors at the beginning of hospitalization can play a crucial role in promoting proper management in the treatment of patients with neutropenia.

Evaluation of lymphocyte subtypes in COVID-19 patients

Background: Although the many aspects of COVID-19 have not been yet recognized, it seems that the dysregulation of the immune system has a very important role in the progression of the disease. In this study the lymphocyte subsets were evaluated in COVID-19 patients with different severity. Methods: In this prospective study, the levels of peripheral lymphocyte subsets (CD3+, CD4+, CD8+ T cells; CD19+ and CD20+ B cells; CD16+/CD56+ NK cells, and CD4+/CD25+/FOXP3+ regulatory T cells) were measured in 67 confirmed patients with COVID-19 on the first day of admission. Results: The mean age of cases was 51.3 plus-or-minus sign 14.8 years. Thirty-two patients (47.8%) were classified as severe cases and 11 (16.4%) patients were categorized as critical. The frequency of blood lymphocytes, CD3+ cells, CD25+FOXP3+ T cells; and absolute count of CD3+ T cells, CD25+FOXP3+ T cells, CD4+ T cells, CD8+ T cells, CD16+56+ lymphocytes were lower in more severe cases in comparison to milder cases. Percentages of lymphocytes, T cells, and NK cells were significantly lower inthe patients who died (p= 0.002 and P= 0.042, p=0.006, respectively). Conclusion: Findings of this cohort study suggests that the frequency of CD4+, CD8+, CD25+FOXP3+ T cells, and NK cells were difference in the severe COVID-19 patients. Moreover, lower frequency of , T cells, and NK cells are predictors of mortality of these patients.

cfDNA in pancreatic neuroendocrine carcinoma management with Cushing’s syndrome

We report a case of metastatic pancreatic neuroendocrine carcinoma associated with paraneoplastic Cushing’s syndrome, successively treated with five lines of treatment (platin-etoposide, LV5FU2-dacarbazine, Folfirinox, pembrolizumab, and paclitaxel) and anti-secretory treatment. Circulating-free DNA (cfDNA) was analysed at each morphological evaluation starting from the second-line treatment. cfDNA changes were well-correlated with the disease course, and cfDNA may be used as a predictive marker and/or an early marker of response. In addition, absolute count of atypical cells was elevated upon disease progression.

An analysis of eight family cluster cases in Wuhan, China: Peripheral blood lymphocyte count may predict Covid-19 mortality

Background: Since December 2019, a novel coronavirus (SARSCoV-2) causing COVID-19 has spread across the world in a global pandemic. Tens of thousands of people were infected, several thousand patients died. However, key risk factors for predicting mortality remain unclear. This study aims to analyze the differences in mortal risk factors between fatal and non-fatal cases within each family, to identify the key risk factors for COVID-19 mortality. Method: Retrospective, randomly selected eight family clusters consisting of 21 individual cases who had been confirmed positive for SARS-CoV-2 and admitted to the Wuhan Union Hospital, Wuhan, China, from February 6 to March 3, 2020. Clinical characteristics and demographic data were tracked up to March 3. Results: Among all 8 family cluster cases, 4 families had death cases. All deaths were elderly individuals (range, 77-88 years), all ICU and severe cases were also elderly individuals (72-88 years). Patient 2-M1, who was the oldest of all cases and first confirmed with COVID-19 on January 10, had four critical comorbid conditions including colon cancer, COPD, hypertension, and coronary disease. But he remains in stable condition after more than 50 days of inpatient treatment. We observed that the absolute count of peripheral blood lymphocyte dropped to less than 0.8G/L of all death and ICU cases, ranging from 0.22 G/L to 0.81 G/L. Conclusions: We found that elderly age is one of the main risk factors for mortality, comorbidities were not predictive of mortality due to COVID-19, although they may extend disease duration. Importantly, we discovered that within our study population the absolute count of peripheral blood lymphocyte is a predictive risk factor for mortality due to COVID-19, establishing that it may be a very important factor for judging a patient’s prognosis. Keywords: COVID-19; Family clusters; Mortality; Risk factors.