High Monocytes Research Articles

IMMU-38. DEEP IMMUNOPROFILING OF HUMAN GLIOBLASTOMA (GBM) REVEALS DIFFERENCES IN THE TUMOR IMMUNE CELL INFILTRATE IN PATIENTS WITH HIGH VS. LOW PLASMA CELL-FREE DNA (CFDNA)

Abstract BACKGROUND We have previously demonstrated that high baseline plasma cfDNA concentration is associated with poor survival in patients with newly diagnosed GBM. The mechanism of this association remains unknown. To explore whether differences in the immune landscape between high- vs. low-cfDNA patients may play a role in their divergent clinical outcomes, we phenotyped tumors from patients with high vs. low cfDNA using mass cytometry by time of flight (CyTOF). METHODS We performed CyTOF on frozen tumor infiltrate suspension from a pilot cohort of patients with previously untreated GBM with known baseline plasma cfDNA concentration (Bagley, Clin Cancer Res 2020). CyTOF was used to simultaneously measure expression of 39 molecules related to immune cell lineage, differentiation state, and function. Differences in immune cell infiltrates between high- and low-cfDNA patients were assessed using Mann-Whitney U tests. RESULTS Four patients with high cfDNA (median 57, range 33-90 ng/mL) were compared to six patients with low cfDNA (median 12, range 7-16 ng/mL). Immune cell infiltrates with increased adaptive cells (high monocytes and T cells, p=0.05) were present in high-cfDNA compared to low-cfDNA patients. While > 70% of the infiltrating T cells were exhausted in both groups, the pattern of exhaustion was significantly different in high- vs. low-cfDNA patients, with less CXCR5+CD69+ and more CXCR5-CD69- (p=0.008) progenitor exhausted T cells in cfDNA-high patients. CONCLUSIONS In this GBM pilot study, we demonstrated differences in the tumor immune infiltrate in patients with high vs. low baseline plasma cfDNA concentration. Preclinical studies will be needed to determine if this explains the association between high plasma cfDNA and poor outcomes previously observed in patients. Our results may have implications for the use of cfDNA concentration as a predictive biomarker for immunotherapy, as tumors with more intermediate progenitor (CXCR5-CD69-) exhausted T cells may respond better to PD-1 checkpoint blockade.

The Utility of Extended White Blood Cell Differential by Flow Cytometry in Routine Hematology.

Abstract 4708Today's Hematology instruments identify white blood cells (WBC) by their physical characteristics using various measurements such as Impedance, Conductivity and Light Scatter. A complete cell identification and enumeration is performed but is limited to only six cell types: neutrophils, lymphocytes, monocytes, eosinophils, basophils and NRBCs. Automation of the WBC manual differential is possible by flow cytometry for more accurate and automated information (van Lochem et al. 2004, M. Roussel et al. 2007, J. Feuillard et al. 2007).The CytoDiff” tube is a 5-color / 6-marker panel that provides an extended cytometric differential for whole blood specimens. The CytoDiff” tube is comprised of: CD36-FITC, (CD2+CD294)-PE, CD19-ECD, CD16-PC5, and CD45-PC7 (Beckman Coulter, Inc. (BEC), For Investigational Use Only. The performance characteristics of this product have not been established).The peripheral blood (PB) specimens were lyzed using VersaLyse (BEC) to prevent selective loss of cells and to conserve light scatter characteristics. PB leukocytes were analyzed according to their immunofluorescence reactivity. At least 20,000 CD45+ events per sample were acquired on an FC500 Cytomics flow cytometer (BEC) and analyzed using CXP software (BEC). For each case with elevated numbers of leukocytes or with abnormal cells, additional analyses were performed using light microscopy and full panel extended flow cytometric immunophenotyping to identify lineages and the level of differentiation of the cells. PB was evaluated from 62 patients with different pathological conditions. Half the patients were females and half males, with adult median age 60. Four boys; median age 10.5, and two girls; median age 4.5 were also evaluated. The study was approved by The Pavlov State Medical University's Institutional Review Board, and the following results were obtained:Two cases had no immunological abnormalities in WBC subsets. Extended immunophenotyping determined 13 lymphocytosis cases with more than 50% of nucleated cells (53.1–94.5%) to be B-lymphocytosis, with 12 B-ChLL and one PLL. One specimen with normal lymphocytes had high B-cells (45%) and extended immunophenotyping specified it as hairy cell leukemia.Six cases were identified with B-blasts, nine with noTnoB (myelo) blasts and two with immature granulocytes. The analysis of these specimens using light microscopy, extended immunophenotyping and cytogenetics identified six B-cell precursors ALL, two cases of MDS transformation to AML, two relapses of AML with maturation, one new diagnosed case of AML with maturation, one case of AML with myelomonoblastic differentiation, three cases of residual disease after AML chemotherapy, one plasmoblastic leukemia (transformed after multiple myeloma) and one acute promyelocytic leukemia with t(15;17). Therefore, true positive blasts were identified in15 of 17 PB specimens using extended immunophenotyping. Two cases with immature granulocytes in PB were misclassified with the CytoDiff” tube but verified as plasmablastic and acute promyelocytic leukemia after bone marrow extended immunophenotyping and cytogenetics.Nine cases had increased numbers of monocytes (14.2–32.0%). Three cases came from the surgery department, with more than 20% of monocytes being inflammatory CD16+. Two cases with multiple myeloma after chemotherapy and cytopenia recovery had 38.8% and 43.0% of monocytes being inflammatory CD16+. Another three cases (two with craniocerebral trauma, and one with phlegmona of mandibula) showed very low inflammatory monocytes (3.5–9.3%). There was one case with very high monocytes (32%) but none were inflammatory. The analysis of this specimen using light microscopy, extended immunophenotyping and cytogenetics identified promyelocytic leukemia. ConclusionThese data demonstrate that a differential leukocyte count using Cytodiff” allows for verification of a wide spectrum of normal and pathological cells in peripheral blood. This will provide clinicians with a diagnostic tool to rapidly determine appropriate therapies for a wide variety of blood-related diseases. Disclosures:No relevant conflicts of interest to declare.

MICROENVIRONMENT IMMUNE CELLS IN BONE MARROW AND LYMPH NODE MEASURED BY FLOW CYTOMETRY CAN PREDICT SURVIVAL IN DIFFUSE LARGE B CELL LYMPHOMA

Introduction: Tumor microenvironment (TME) in diffuse large B-cell lymphoma (DLBCL) has recently been in the spotlight. The predictive role of immune cells (IC) in peripheral blood (PB), bone marrow (BM) and lymph nodes (LN) has been individually assessed, often using techniques not widely available. However, data regarding the relative prognostic impact of TME in each compartment evaluated simultaneously in DLBCL is still lacking. Aims: To determine the prognostic impact on survival of the Monocyte-Lymphocyte prognostic score (MLS) in PB and the percentage (%) of IC in the BM and LN measured by flow cytometry (FC) in DLBCL. Methods: We retrospectively collected information of patients (pts) with DLBCL and available BM aspiration and LN FC data at diagnosis, treated at our center between 2012 and 2019. Pts were stratified according to the MLS (Wilcox et al, leukemia 2011) in two groups: low (PB monocyte counts <630/ml and lymphocyte counts ≥1000/ml) and intermediate-high. FC analysis was performed with 8-color panels according to Euroflow protocols. % of BM and LN IC by FC were compared to normal values determined by Matarraz (Cytometry part B 2010) and Battaglia et al (Immunology 2003), and analyzed in 3 categories: low, normal and high. Survival analysis was performed with Kaplan-meier (variables compared with log-rank) and Cox regression. Results: 71 pts were included with a median age of 59 years. 49.3% had poor R-IPI. All pts received immunochemotherapy. Complete remission rate was 82%. Median overall survival (OS) was 120.5 months (m), with a median follow up time of 34.7 m. On univariate analysis TME variables from all the compartments showed prognostic impact on OS. BM IC levels did not statistically differ in involved vs not involved BM. Both low LN TL % and high and low BM Mo % remained independent predictors of survival on multivariate analysis. Moreover, pts with both of these unfavorable variables had a HR for death of 40.22 (CI 95% 5.16-313.04, p < 0.001) with a median OS of only 22.19 m. Variables Univariate analysis Multivariate analysis HR (95% CI) P-value HR (95% CI) P-value LN T lymphocytes (TL) low 3.98 (1.31-12.06) 0.014 5.44 (1.48-20.04) 0.011 BM B lymphocytes low 4.73 (1.62-13.79) 0.004 3.36 (0.99-11,34) 0.0501 BM Monocytes (Mo) High 10.53 (3.66-30.26) <0.001 4.66 (1.34-16.25) 0.015 BM Monocytes Low 3.87 (1.28-11.7) 0.016 3.40 (1.05-10.99) 0.04 PB MLS intermediate-high 2.59 (1.12-6) 0.02 1.05 (0.4-2.77) 0.91 Conclusion: Abnormal BM Mo % and low LN TL % measured by FC were associated with inferior OS in our cohort of DLBCL pts. These two TME variables presented at diagnosis could be easily translated into prognostic biomarkers in DLBCL, identifying a subgroup of pts with a dismal short term prognosis. In the future, therapies that target the crosstalk between lymphoma cells and the LN and BM ME might represent an encouraging strategy to improve outcomes in this high risk subset of pts. Kaplan-meler estimates of overall survival stratified by number of adverse prognosis tumor microenviroment varaibles present(low lymph node T-lymphocytes and high low bone marrow monocytes) Keywords: Diagnostic and Prognostic Biomarkers No conflicts of interests pertinent to the abstract. On univariate analysis TME variables from all the compartments showed prognostic impact on OS. BM IC levels did not statistically differ in involved vs not involved BM. Both low LN TL % and high and low BM Mo % remained independent predictors of survival on multivariate analysis. Moreover, pts with both of these unfavorable variables had a HR for death of 40.22 (CI 95% 5.16-313.04, p<0.001) with a median OS of only 22.19 m. Variables Univariate analysis Multivariate analysis HR (95% CI) P-value HR (95% CI) P-value LN T lymphocytes (TL) low 3.98 (1.31-12.06) 0.014 5.44 (1.48-20.04) 0.011 BM B lymphocytes low 4.73 (1.62-13.79) 0.004 3.36 (0.99-11,34) 0.0501 BM Monocytes (Mo) High 10.53 (3.66-30.26) <0.001 4.66 (1.34-16.25) 0.015 BM Monocytes Low 3.87 (1.28-11.7) 0.016 3.40 (1.05-10.99) 0.04 PB MLS intermediate-high 2.59 (1.12-6) 0.02 1.05 (0.4-2.77) 0.91

The Prognostic Impact of Eosinophils and the Eosinophil-to-Lymphocyte Ratio on Survival Outcomes in Stage II Resectable Pancreatic Cancer.

The relationship between eosinophils and cancer prognosis is unknown. Therefore, we analyzed the relationship between circulating eosinophils and the survival of stage IIA and IIB pancreatic cancer patients who underwent surgical resection. This study included a retrospective cohort of 67 consecutive patients. Patients were categorized into two different groups based on the optimal cutoff for pretreatment levels of each biomarker, according to the receiver operating characteristic curves. The Kaplan-Meier method showed that low eosinophil (P = 0.0403), high neutrophil (P = 0.0066), and high monocyte (P = 0.0003) counts were associated with short overall survival (OS). Low lymphocyte-to-monocyte ratio (P = 0.0194) and eosinophil-to-lymphocyte ratio (ELR) (P = 0.0413) were associated with reduced OS. In multivariate analysis, histological differentiation (P = 0.0014), high neutrophils (P = 0.047), high monocytes (P = 0.029), and low eosinophils (P < 0.0001) were correlated with poorer OS. Histological differentiation (P = 0.033), low lymphocyte-to-monocyte ratio (P = 0.029), and low ELR (P = 0.005) were correlated with poor OS and were significant independent prognostic factors of poor outcomes. Low eosinophils and low ELR were significant independent prognostic factors of poor outcomes.

Deciphering the complex circulating immune cell microenvironment in chronic lymphocytic leukaemia using patient similarity networks

The tissue microenvironment in chronic lymphocytic leukaemia (CLL) plays a key role in the pathogenesis of CLL, but the complex blood microenvironment in CLL has not yet been fully characterised. Therefore, immunophenotyping of circulating immune cells in 244 CLL patients and 52 healthy controls was performed using flow cytometry and analysed by multivariate Patient Similarity Networks (PSNs). Our study revealed high inter-individual heterogeneity in the distribution and activation of bystander immune cells in CLL, depending on the bulk of the CLL cells. High CLL counts were associated with low activation on circulating monocytes and T cells and vice versa. The highest activation of immune cells, particularly of intermediate and non-classical monocytes, was evident in patients treated with novel agents. PSNs revealed a low activation of immune cells in CLL progression, irrespective of IgHV status, Binet stage and TP53 disruption. Patients with high intermediate monocytes (> 5.4%) with low activation were 2.5 times more likely (95% confidence interval 1.421–4.403, P = 0.002) to had shorter time-to-treatment than those with low monocyte counts. Our study demonstrated the association between the activation of circulating immune cells and the bulk of CLL cells. The highest activation of bystander immune cells was detected in patients with slow disease course and in those treated with novel agents. The subset of intermediate monocytes showed predictive value for time-to-treatment in CLL.

Peripheral Blood Smear Findings of COVID-19 Patients Provide İnformation about the Severity of the Disease and the Duration of Hospital Stay.

BackgroundData about the morphological changes in peripheral blood smears during COVID-19 infection and their clinical severity association are limited. We aimed to examine the characteristics of the cells detected in the pathological rate and/or appearance and whether these findings are related to the clinical course by evaluating the peripheral blood smear at the time of diagnosis in COVID-19 patients.MethodsClinical features, laboratory data, peripheral blood smear of fifty patients diagnosed with COVID-19 by PCR was evaluated at diagnosis. Peripheral smear samples of the patients were compared with the age and sex-matched 30 healthy controls. Pictures were taken from the patient’s peripheral blood smear. Patients were divided into two groups. Mild and severe stage patient groups were compared in terms of laboratory data and peripheral smear findings. The relationship between the laboratory values of all patients and the duration of hospitalization was analyzed.ResultsThe number of segmented neutrophils and eosinophils were low, pseudo-Pelger-Huet, pseudo-Pelger-Huet/mature lymphocyte ratio, atypical lymphocytes, monocytes with vacuoles, bands, and pyknotic neutrophils rates were higher in the peripheral blood smear of the patient group (p <0.05). Increased pseudo-Pelger-Huet anomaly, pseudo-Pelger Huet/mature lymphocyte ratio, a decreased number of mature lymphocytes, and eosinophils in peripheral blood smear were observed in the severe stage patients (p <0.05). A negative correlation was observed between hospitalization duration and mature lymphocyte and monocytes with vacuoles rates (p <0.05).ConclusionA peripheral blood smear is an inexpensive, easily performed, and rapid test. Increased Pseudo-Pelger-Huet anomaly/mature lymphocyte rate suggests a severe stage disease, while high initial mature lymphocyte and monocytes with vacuoles rates at the time of diagnosis may be an indicator of shortened duration of hospitalization.

Low High-Density Lipoprotein Cholesterol and High White Blood Cell Counts: A Mendelian Randomization Study.

Animal studies suggest that HDL (high-density lipoprotein) regulates proliferation and differentiation of hematopoietic stem cells. Using a Mendelian randomization approach, we tested the hypothesis that low HDL cholesterol is associated with high white blood cell counts. Approach and Results: We included 107 952 individuals aged 20 to 100 years from the Copenhagen General Population Study with information on HDL cholesterol, white blood cell counts, and 9 genetic variants associated with HDL cholesterol. In multivariable-adjusted observational analyses, HDL cholesterol was inversely associated with white blood cell counts. On a continuous scale, a 1-mmol/L (39 mg/dL) lower HDL cholesterol was associated with 5.1% (95% CI, 4.7%-5.4%) higher leukocytes, 4.5% (95% CI, 4.0%-4.9%) higher neutrophils, 5.7% (95% CI, 5.3%-6.1%) higher lymphocytes, 5.7% (95% CI, 5.3%-6.2%) higher monocytes, 14.8% (95% CI, 13.9%-15.8%) higher eosinophils, and 3.9% (95% CI, 3.1%-4.7%) higher basophils. In age- and sex-adjusted genetic analyses using the inverse-variance weighted analysis, a 1-mmol/L (39 mg/dL) genetically determined lower HDL cholesterol was associated with 2.2% (95% CI, 0.3%-4.1%) higher leukocytes, 4.3% (95% CI, 1.6%-7.1%) higher lymphocytes, 4.3% (95% CI, 2.6%-6.1%) higher monocytes, and 4.8% (95% CI, 1.2%-8.5%) higher eosinophils. Overall, the genetic associations were robust across sensitivity analyses and replicated using summary statistics from the UK Biobank with up to 350 470 individuals. Genetic and hence lifelong low HDL cholesterol was associated with high peripheral blood leukocytes, including high lymphocytes, monocytes, and eosinophils. The concordance between observational and genetic estimates and independent replication suggest a potential causal relationship.

Simulation of atherosclerotic plaque growth using computational biomechanics and patient-specific data

Atherosclerosis is the one of the major causes of mortality worldwide, urging the need for prevention strategies. In this work, a novel computational model is developed, which is used for simulation of plaque growth to 94 realistic 3D reconstructed coronary arteries. This model considers several factors of the atherosclerotic process even mechanical factors such as the effect of endothelial shear stress, responsible for the initiation of atherosclerosis, and biological factors such as the accumulation of low and high density lipoproteins (LDL and HDL), monocytes, macrophages, cytokines, nitric oxide and formation of foams cells or proliferation of contractile and synthetic smooth muscle cells (SMCs). The model is validated using the serial imaging of CTCA comparing the simulated geometries with the real follow-up arteries. Additionally, we examine the predictive capability of the model to identify regions prone of disease progression. The results presented good correlation between the simulated lumen area (P < 0.0001), plaque area (P < 0.0001) and plaque burden (P < 0.0001) with the realistic ones. Finally, disease progression is achieved with 80% accuracy with many of the computational results being independent predictors.

Abstract B25: Demographic characteristics and peripheral blood clinical laboratory variables predict irAE occurrence in patients with advanced melanoma receiving anti-PD-1 monotherapy

Abstract Background: Although immune-related adverse effects (irAEs) are commonly seen in anti-PD-1 monotherapy-treated patients, their predictive biomarkers are still lacking. Methods: In this retrospective analysis, clinical data were collected from 141 advanced melanoma patients treated with anti-PD-1 monotherapy at Massachusetts General Hospital from Sept. 2009 to Dec. 2017. Demographic characteristics, baseline, and early-on-treatment (median 3 weeks after anti-PD-1 monotherapy initiation) routine laboratory variables were collected and correlated with occurrence of irAEs in both univariate and multivariate analyses. Results: 80/141 (56.7%) patients developed irAEs after anti-PD-1 monotherapy initiation. Significant independent predictive variables (P &amp;lt; 0.05) in favor of the occurrence of irAEs included the following: with no distant metastasis (Stage M0) before treatment initiation, high baseline neutrophils, and high early-on-treatment neutrophil-to-lymphocyte ratio (NLR). High baseline monocytes, high baseline basophils, and high early-on-treatment WBCs were of borderline significance (P &amp;lt; 0.1) in multivariate analyses. An irAE risk scoring model (total scale: 0-6) comprising these 6 factors (all as dichotomous variables) was established (HR 1.781, 95% CI 1.450 – 2.187, P &amp;lt; 0.001). Risks of developing at least one irAE within 12 weeks after anti-PD-1 monotherapy initiation were 0%, 21%, 21%, 39%, 30%, and 63%; within 48 weeks were 0%, 41%, 57%, 68%, 77%, and 100%, for patients with scores of 0 to 5 (no patient scored 6 within this cohort), respectively. Conclusions: Scoring system based on simple, easily accessible, and routinely tested biomarkers could be used to help predict the risk of irAEs occurrence in anti-PD-1 monotherapy-treated advanced melanoma patients. Citation Format: Xue Bai, Michelle S. Kim, Gyulnara Kasumova, Justine Cohen, Donald Lawrence, Christine Freedman, Riley Fadden, Krista Rubin, Tatyana Sharova, Keith Flaherty, Ryan Sullivan, Genevieve Boland. Demographic characteristics and peripheral blood clinical laboratory variables predict irAE occurrence in patients with advanced melanoma receiving anti-PD-1 monotherapy [abstract]. In: Proceedings of the AACR Special Conference on Melanoma: From Biology to Target; 2019 Jan 15-18; Houston, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(19 Suppl):Abstract nr B25.

The Role of ROCK in Platelet-Monocyte Collaborative Induction of Thromboinflammation during Acute Coronary Syndrome.

Arterial thrombosis is initiated by atherosclerotic plaque damage, prothrombotic material release and platelet aggregation. Platelets are primary mediators involved in thrombosis and cooperate with vascular and immune cells. Herein, we investigated how activated platelets interacted with monocytes in atherothrombosis. We collected patients' blood from coronary arteries during percutaneous coronary intervention and measured platelet activity. Platelets from coronary arteries had higher pseudopodium expression and activity in patients with acute coronary syndrome (ACS). Ribosome profiling of platelets from coronary blood mapped a vigorous upregulation of Rho GTPases and their downstream effectors. RhoA activated downstream Rho-associated coiled-coil containing protein kinase (ROCK), and ROCK increased surface P-selectin in coronary blood platelets. The interaction between platelets and monocytes was observed in vitro, and was found in ruptured coronary plaques of ACS. Further we found that activated platelets promoted monocytes transmigration, which could be suppressed in the presence of ROCK inhibitors. The increased surface P-selectin on thrombin-induced platelets interacted with monocytes to upregulate monocyte chemokine receptor 2 (CCR2) expression via the ROCK pathway. The expression of CCR2 was higher in monocyte-platelet aggregates than in monocytes without platelets. Finally, using the Asian Screening Array BeadChip, we identified single-nucleotide polymorphism (SNP) associated with cardiovascular events. Notably, patients having homozygous major alleles of the RHOA SNP rs11706370 presented with higher risks of cardiovascular events. Through ROCK-activated cytoskeleton remodeling and P-selectin expression, platelets were recruited and interacted synergistically with high CCR2-expressing monocytes to induce thromboinflammation in atherothrombosis.

The Metabolic Changes and Immune Profiles in Patients With COVID-19.

To explore the metabolic changes and immune profiles in patients with COVID-19, we analyzed the data of patients with mild and severe COVID-19 as well as young children with COVID-19. Of the leukocytes, 47% (IQR, 33–59) were lymphocytes [2.5 × 109/L (IQR, 2.2–3.3)], and monocytes were 0.51 × 109/L (IQR, 0.45–0.57) in young children with COVID-19. In 32 mild COVID-19 patients, circulating monocytes were 0.45 × 109/L (IQR, 0.36–0.64). Twenty-one severe patients had low PO2 [57 mmHg (IQR, 50–73)] and SO2 [90% (IQR, 86–93)] and high lactate dehydrogenase [580 U/L (IQR, 447–696)], cardiac troponin I [0.07 ng/mL (IQR, 0.02–0.30)], and pro-BNP [498 pg/mL (IQR, 241–1,726)]. Serum D-dimer and FDP were 9.89 mg/L (IQR, 3.62–22.85) and 32.7 mg/L (IQR, 12.8–81.9), and a large number of RBC (46/μL (IQR, 4–242) was presented in urine, a cue of disseminated intravascular coagulation (DIC) in severe patients. Three patients had comorbidity with diabetes, and 18 patients without diabetes also presented high blood glucose [7.4 mmol/L (IQR, 5.9–10.1)]. Fifteen of 21 (71%) severe cases had urine glucose +, and nine of 21 (43%) had urine ketone body +. The increased glucose was partially caused by reduced glucose consumption of cells. Severe cases had extraordinarily low serum uric acid [176 μmol/L (IQR, 131–256)]. In the late stage of COVID-19, severe cases had extremely low CD4+ T cells and CD8+ T cells, but unusually high neutrophils [6.5 × 109/L (IQR, 4.8–9.6)], procalcitonin [0.27 ng/mL (IQR, 0.14–1.94)], C-reactive protein [66 mg/L (IQR, 25–114)] and an extremely high level of interleukin-6. Four of 21 (19%) severe cases had co-infection with fungi, and two of 21 (9%) severe cases had bacterial infection. Our findings suggest that, severe cases had acute respiratory distress syndrome (ARDS) I–III, and metabolic disorders of glucose, lipid, uric acid, etc., even multiple organ dysfunction (MODS) and DIC. Increased neutrophils and severe inflammatory responses were involved in ARDS, MODS, and DIC. With the dramatical decrease of T-lymphocytes, severe cases were susceptible to co-infect with bacteria and fungi in the late stage of COVID-19. In young children, extremely high lymphocytes and monocytes might be associated with the low morbidity of COVID-19. The significantly increased monocytes might play an important role in the recovery of patients with mild COVID-19.

An engineered anti-idiotypic antibody-derived killer peptide (KP) early activates swine inflammatory monocytes, CD3+CD16+ natural killer T cells and CD4+CD8α+ double positive CD8β+ cytotoxic T lymphocytes associated with TNF-α and IFN-γ secretion

This study evaluated the early modulation of the phenotype and cytokine secretion in swine immune cells treated with an engineered killer peptide (KP) based on an anti-idiotypic antibody functionally mimicking a yeast killer toxin. The influence of KP on specific immunity was investigated using porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) as ex vivo antigens. Peripheral blood mononuclear cells (PBMC) from healthy pigs were stimulated with KP and with a scramble peptide for 20 min, 1, 4 and 20 h or kept unstimulated. The cells were analyzed using flow cytometry and ELISA. The same time-periods were used for KP pre-incubation/co-incubation to determine the effect on virus-recalled interferon-gamma (IFN-γ) secreting cell (SC) frequencies and single cell IFN-γ productivity using ELISPOT.KP induced an early dose-dependent shift to pro-inflammatory CD172α+CD14+high monocytes and an increase of CD3+CD16+ natural killer (NK) T cells. KP triggered CD8α and CD8β expression on classical CD4−CD8αβ+ cytotoxic T lymphocytes (CTL) and double positive (DP) CD4+CD8α+ Th memory cells (CD4+CD8α+low CD8β+low). A fraction of DP cells also expressed high levels of CD8α. The two identified DP CD4+CD8α+high CD8β+low/+high CTL subsets were associated with tumor necrosis factor alpha (TNF-α) and IFN-γ secretion. KP markedly boosted the reactivity and cross-reactivity of PRRSV type-1- and PCV2b-specific IFN-γ SC. The results indicate the efficacy of KP in stimulating Th1-biased immunomodulation and support studies of KP as an immunomodulator or vaccine adjuvant.

Aberrant activation of bone marrow Ly6Chighmonocytes in diabetic mice contributes to impaired glucose tolerance.

Accumulating evidence indicates that diabetes and obesity are associated with chronic low-grade inflammation and multiple organ failure. Tissue-infiltrated inflammatory M1 macrophages are aberrantly activated in these conditions and contribute to hyperglycemia and insulin resistance. However, it is unclear at which stage these cells become aberrantly activated: as precursor monocytes in the bone marrow or as differentiated macrophages in tissues. We examined the abundance, activation state, and function of bone marrow-derived Ly6Chigh monocytes in mice with diabetes and/or obesity. Ly6Chigh monocytes were FACS-purified from six groups of male mice consisting of type 2 diabetes model db/db mice, streptozotocin (STZ) induced insulin depletion mice, high fat diet (HFD) induced obesity mice and each control mice. Ly6Chigh monocytes were then analyzed for the expression of inflammation markers by qRT-PCR. In addition, bone marrow-derived Ly6Chigh monocytes from db/+ and db/db mice were fluorescently labeled and injected into groups of db/db recipient mice. Cell trafficking to tissues and levels of markers were examined in the recipient mice. The expression of many inflammation-related genes was significantly increased in Ly6Chigh monocytes from db/db mice, compared with the control. Bone marrow-derived Ly6Chigh monocytes isolated from db/db mice, but not from db/+ mice, displayed prominent infiltration into peripheral tissues at 1 week after transfer into db/db mice. The recipients of db/db Ly6Chigh monocytes also exhibited significantly increased serum glucose levels and worsening tolerance compared with mice receiving db/+ Ly6Chigh monocytes. These novel observations suggest that activated Ly6Chigh monocytes may contribute to the glucose intolerance observed in diabetes.

EXAMINATION OF PULMONARY TUBERCULOSIS PATIENTS AT THE REGIONAL GENERAL HOSPITAL (RSUD) JAYAPURA

Tuberculosis is a disease that is directly transmitted and caused by bacteria Mycobacterium tuberculosis. Tuberculosis is an infectious disease that still causes high morbidity and mortality rates in the world. This research aims to determine the value of hemoglobin levels, platelet counts, erythrocytes, the number of neutrophils and Monocytes in positive patients with pulmonary tuberculosis who performed examinations at the Jayapura Regional General Hospital. This research was conducted for 1 month, starting from 1 May to 1 June 2018. This research is descriptive with a laboratory test approach. The population in this research included all patients who had positive pulmonary tuberculosis and the samples in this study were venous blood positive patients with pulmonary tuberculosis. The method used in this examination is flow cytometry using the hematology analyzer Sysmex XN-1000. The results of the research showed that from 20 samples, hemoglobin levels were obtained as much as 30% with normal hemoglobin levels and as much as 70% with low hemoglobin levels 70% of normal platelet value and as much as 30% high platelet count; 70% of the value of erythrocytes is normal and as much as 30% the amount of erythrocytes decreases; Neutrophils are as high as 55% while normal is 45%; and high monocytes as much as 25% and normal monocytes as much as 75%.

New prognosis score including absolute lymphocyte/monocyte ratio, red blood cell distribution width and beta-2 microglobulin in patients with diffuse large B-cell lymphoma treated with R-CHOP: Spanish Lymphoma Group Experience (GELTAMO).

The International Prognostic Index (IPI) is the most widely used score for non-Hodgkin lymphoma but lacks the ability to identify a high-risk population in diffuse large B-cell lymphoma (DLBCL). Low absolute lymphocyte count and high monocytes have proved to be unfavourable factors. Red-cell distribution width (RDW) has been associated with inflammation and beta-2 microglobulin (B2M) with tumour load. The retrospective study included 992 patients with DLBCL treated with R-CHOP. In the multivariate analysis, age, Eastern Cooperative Oncology Group performance status (ECOG-PS), stage, bulky mass, B2M, RDW, and lymphocyte/monocyte ratio (LMR) were independently related to progression-free survival (PFS). A new prognosis score was generated with these variables including age categorized into three groups (0, 1, 2 points); ECOG ≥3-4 with two; stage III/IV, bulky mass, high B2M, LMR<2·25 and RDW>0·96 with one each; for a maximum of 9. This score could improve the discrimination of a very high-risk subgroup with five-year PFS and overall survival (OS) of 19% and 24% versus 45% and 59% of R (revised)-IPI respectively. This score also showed greater predictive ability than IPI. A new score is presented including complete blood cell count variables and B2M, which are readily available in real-life practice without additional tests. Compared to R-IPI, it shows a more precise high-risk assessment and risk discrimination for both PFS and OS.

Innate Immune Cells and C-Reactive Protein in Acute First-Episode Psychosis and Schizophrenia: Relationship to Psychopathology and Treatment

Innate immunity has been linked to initiation of Alzheimer’s disease and multiple sclerosis. Moreover, risk of first-episode psychosis (FEP) and schizophrenia (Sz) is increased after various infections in predisposed individuals. Thus, we hypothesized an analogous role of innate immunity with increased C-reactive protein (CRP) in non-affective psychosis. Differential blood count, CRP, neutrophil and monocyte–macrophage activation markers, cortisol and psychotic symptoms (Positive and Negative Syndrome Scale [PANSS]) were assessed in controls (n = 294) and acutely ill unmedicated FEP (n = 129) and Sz (n = 124) patients at baseline and after 6 weeks treatment. Neutrophils, monocytes, and CRP were increased in patients vs controls at baseline (P < .001), and neutrophil and monocyte counts correlated positively with activation markers. Eosinophils were lower at baseline in FEP (P < .001) and Sz (P = .021) vs controls. Differences in neutrophils (P = .023), eosinophils (P < .001), and CRP (P < .001) were also present when controlling for smoking and cortisol, and partially remitted after antipsychotic treatment. FEP patients with high neutrophils (P = .048) or monocytes (P = .021) had higher PANSS-P scores at baseline but similar disease course. CRP correlated with PANSS-P at baseline (ρ = 0.204, P = .012). Improvement of positive symptoms after treatment correlated with declining neutrophils (ρ = 0.186, P = .015) or CRP (ρ = 0.237, P = .002) and rising eosinophils (ρ = −0.161, P = .036). In FEP, normalization of neutrophils (ρ = −0.231, P = .029) and eosinophils (ρ = 0.209, P = .048) correlated with drug dosage. In conclusion, innate immune system activation correlated with PANSS-P, supporting the immune hypothesis of psychosis. Neutrophil and monocyte counts and CRP levels may be useful markers of disease acuity, severity, and treatment response.

A computational multi-level atherosclerotic plaque growth model for coronary arteries.

In this work, we present a novel computational approach for the prediction of atherosclerotic plaque growth. In particular, patient-specific coronary computed tomography angiography (CCTA) data were collected from 60 patients at two time points. Additionally, blood samples were collected for biochemical analysis. The CCTA data were used for 3D reconstruction of the coronary arteries, which were then used for computational modeling of plaque growth. The model of plaque growth is based on a multi-level approach: i) the blood flow is modeled in the lumen and the arterial wall, ii) the low and high density lipoprotein and monocytes transport is included, and iii) the major atherosclerotic processes are modeled including the foam cells formation, the proliferation of smooth muscle cells and the formation of atherosclerotic plaque. Validation of the model was performed using the follow-up CCTA. The results show a correlation of the simulated follow-up arterial wall area to be correlated with the corresponding realistic follow-up with r2=0.49, P<; 0.0001.

CLINICAL NUTRITION PROGRAM IMPROVES FATIGUE, ANXIETY, BRAIN FOG AND IMMUNO-HORMONAL AND GASTROINTESTINAL SYMPTOMS IN A 40 YEAR OLD FEMALE WITH SYSTEMIC ERYTHEMATOUS LUPUS

Event Abstract Back to Event CLINICAL NUTRITION PROGRAM IMPROVES FATIGUE, ANXIETY, BRAIN FOG AND IMMUNO-HORMONAL AND GASTROINTESTINAL SYMPTOMS IN A 40 YEAR OLD FEMALE WITH SYSTEMIC ERYTHEMATOUS LUPUS Tatiana Habanova1* 1 The Pilates Wellness Center, United States Background: A 4o year old female presented with fatigue and anxiety in maintaining her work schedule as a dietician while caring for her two young children and living an active life. She required daily naps, many times working from home in bed. She had constant brain fog and difficulty finding words. She was often “under the weather” taking months to recover. She experienced painful menstrual cramps two weeks prior to menses which were themselves irregular. Constant irritable bowel symptoms, caused her to use activated charcoal to cope with severe bloating and discomfort. Strict sun avoidance was necessary otherwise a rash developed and she expressed great concern over her progressive diffuse patchy hair loss. A recent diagnosis of Systemic Erythematous Lupus (SLE) was made by a rheumatologist but she states her symptoms have been occurring for 2 years prior to diagnosis. No treatment was prescribed at time of diagnosis. Her diet was unrestricted, she drank red wine or light beer every other day, does not smoke, had exercise intolerance, slept well yet woke up fatigued, surgical history significant for cholecystectomy and a family history significant for gastrointestinal cancers. Methods: The patient had a strong adverse clinical reaction to Th-2 stimulators. Blood chemistry revealed lab low Total Cholesterol (131 mg/dl); functionally low Vitamin D (36 ng/ml); lab high CRP (5.6 mg/l), AST (49 U/L) and ALT (62 U/L); functionally high Monocytes (10.3%). She was placed on an anti-inflammatory diet and supplement protocol including sublingual vitamin D, turmeric, resveratrol, glutamine and multi-vitamin/mineral with therapeutic levels of chromium and vanadium. Results: After 120 days on her treatment plan, the patient reported significant improvements: hormonal and gastrointestinal symptoms, as well as patchy hair, lose resolved 100%; fatigue improved 95% with now only needing one nap per month and she now distinguishes feeling tired vs feeling fatigued and adjusts her activities accordingly to avoid “crashing”; she no longer experiences exercise intolerance and started gentle exercise four times per week without negative consequences; she was ill twice during her treatment plan both episodes resolved within two weeks; she can tolerate brief sun exposure (3-5 min) without a rash developing; and states her anxiety has reduced by 90% as she feels capable of living an active life and juggling her work schedule with family time while successfully managing her SLE symptoms. Conclusions: The author suggests further investigation into clinical nutrition and supplementation in the treatment of fatigue, anxiety, brain fog, and immune-hormonal and gastrointestinal symptoms, in patients with a history of SLE. The research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. Keywords: Autoimmunity, Systemic erythematous lupus, supplementation, Anti-inflammatory diet, Fatigue Conference: International Symposium on Clinical Neuroscience, Orlando, United States, 24 May - 26 May, 2019. Presentation Type: Poster Presentation Topic: Clinical Neuroscience Citation: Habanova T (2019). CLINICAL NUTRITION PROGRAM IMPROVES FATIGUE, ANXIETY, BRAIN FOG AND IMMUNO-HORMONAL AND GASTROINTESTINAL SYMPTOMS IN A 40 YEAR OLD FEMALE WITH SYSTEMIC ERYTHEMATOUS LUPUS. Front. Neurol. Conference Abstract: International Symposium on Clinical Neuroscience. doi: 10.3389/conf.fneur.2019.62.00072 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 09 Apr 2019; Published Online: 27 Sep 2019. * Correspondence: Dr. Tatiana Habanova, The Pilates Wellness Center, Wellington, United States, drhabanova@yahoo.com Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Tatiana Habanova Google Tatiana Habanova Google Scholar Tatiana Habanova PubMed Tatiana Habanova Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Abstract 610: Functional Relationship of The CARS2 Locus to Coronary Artery Disease

In a recent meta-analysis of GWAS for cardiovascular disease (CAD) with imputation from 1000 Genomes (Nat Genet 2015), we identified 202 significant CAD associated variants (FDR q&lt; 0.05) in 129 loci. Seven independent association signals were identified in a gene cluster at chromosome 13q34 including rs61969072. This SNP is located in an intergenic region proximal to the ING1 , CARKD and CARS2 genes. Expression data (eQTL) obtained from the GTEx Project demonstrate a strong association of rs61969072 with CARS2 mRNA abundance in artery with decreased CARS2 expression in carriers of the risk G allele, hinting that CARS2 may be a causal CAD gene. CARS2 encodes a putative mitochondrial cysteinyl t-RNA synthetase. Bioinformatics data (BioGPS) indicate high gene expression of CARS2 in monocytes, a precursor of macrophages. Having then demonstrated that CARS2 mRNA expression is decreased in M1 macrophages compared to M0 and M2 macrophages, we examined the role of CARS2 in the anti-inflammatory pathway in macrophages using THP-1 cells, a macrophage model system. Gene expression profiling of 84 key players of the inflammatory response was performed on siRNA mediated knockdown of CARS2 samples. Comparisons between knockdown and control samples (n=4) identified 12 upregulated and 4 down regulated genes with ≥ 1.25 or ≤0.75 fold difference in expression (p&lt;0.05). Gene Set Enrichment Analysis (GSEA), using inflammatory response PCR array data, identified the interleukin-10 signaling pathway as an enriched category (FDR q&lt;0.05). IL-10 has an atheroprotective effect against plaque rupture and thrombus formation by influencing the local inflammatory process in the lesion. IL-10 activates the STAT3 cascade, where phosphorylated STAT3 homodimers translocate to the nucleus to activate specific target effector genes which repress the proinflammatory genes at the transcriptional level. IL-10 induced STAT3 phosphorylation was measured by Western blotting in THP-1 cells. In response to IL-10, CARS2 knockdown samples showed dampened STAT3 phosphorylation compared to control suggesting that CARS2 serves an anti-inflammatory function within the IL-10 signaling pathway.

Clinical features of pediatric tuberculous pleurisy

Objective To sum up the clinical manifestations and laboratory features for the diagnosis of pe-diatric tuberculous pleurisy, and to improve the recognition of this disease in early stage. Methods A retrospective study of 113 children diagnosed as tuberculous pleurisy from August 2006 to September 2014 in the Second Department of Respiratory Medicine, Beijing Children′s Hospital Affiliated to Capital Medical University was conducted.Meanwhile, another 113 cases of children with mycoplasma pneumoniae pneumonia complicated with pleurisy were selected as control group.The general information, clinical symptoms, pleural effusion and imaging features between 2 groups were analyzed and compared by using SPSS 16.0 statistical software. Results The proportion of patients with cough in tuberculous pleurisy group and control group was 47.79% (54/113 cases) and 99.12% (112/113 cases) (χ2=76.33, P<0.01) respectively, and the proportion with severe cough was 3.70% (2/54 cases) and 97.32% (109/112 cases) (χ2=144.10, P<0.01), while the disease duration was 15.00 (10.00, 30.00)days and 10.00 (8.00, 14.50)days (W=8 668.00, P<0.01), respectively, and all the differences between 2 groups were significant.The proportion of patients with low fever, moderate fever, high fever and hyper fever was 8.65% (9/104 cases), 47.12% (49/104 cases), 44.23% (46/104 cases) and 0, respectively in tuberculous pleurisy group, while the proportion was 0.90% (1/111 cases), 18.92% (21/111 cases), 79.28% (88/111 cases) and 0.90% (1/111 cases) respectively in control group, and the difference between 2 groups was significant(W=9 064.00, P<0.01). The unilateral effusion ratio in tuberculous pleurisy group and the control group was 94.69% (107/113 cases) and 71.68% (81/113 cases), respectively (χ2=21.39, P<0.01). The monocyte ratio was higher in tuberculous pleurisy group [0.89(0.76, 0.93)] than that in the control group [0.60(0.30, 0.78)](W=888.50, P<0.01)and the level of protein in 2 groups was [51.00(47.35, 54.20)g/L] and [42.10 (37.85, 46.15)g/L], respectively (W=842.50, P<0.01). The level of lactate dehydrogenase (LDH) in tuberculous pleurisy group[553.50 (358.00, 749.25)U/L] was lower than that in the control group[1 189.10 (670.95, 1 820.00) U/L] (W=2 186.00, P<0.01), and the differences were significant between 2 groups.In addition to pleural effusion, the high density was the main feature of imaging examination in 2 groups.The proportion of patients with atelectasis was 77.88% (88/113 cases) and 4.42%(5/113 cases)(χ2=125.90, P<0.01), while the proportion of patients with consolidation was 4.42% (5/113 cases) and 72.57% (82/113 cases), respectively (χ2=110.80, P<0.01). All the differences between 2 groups were significant.The sputum culture-positive rate of mycobacterium tuberculosis was only 1.77% (2/113 cases) and the other pathogen examinations were negative in tuberculous pleurisy group. Conclusions For patients with unilateral pleural effusion, when the onset only has fever (moderate-high fever), and respiratory symptoms are not clear or symptoms are not proportional to radiographic severity, or when high monocytes proportion(>0.70)in pleural effusion and radiographic evidence of compression atelectasis are observed, tuberculous pleurisy should be considered and further questioning of the predisposing factors, purified protein derivative test should be taken so as to diagnose the tuberculous pleurisy at early stage. Key words: Child; Tuberculous pleurisy; Diagnosis