Lower Monocyte Research Articles

Complete blood count as a biomarker for preeclampsia with severe features diagnosis: a machine learning approach

ObjectiveThis study introduces the complete blood count (CBC), a standard prenatal screening test, as a biomarker for diagnosing preeclampsia with severe features (sPE), employing machine learning models.MethodsWe used a boosting machine learning model fed with synthetic data generated through a new methodology called DAS (Data Augmentation and Smoothing). Using data from a Brazilian study including 132 pregnant women, we generated 3,552 synthetic samples for model training. To improve interpretability, we also provided a ridge regression model.ResultsOur boosting model obtained an AUROC of 0.90±0.10, sensitivity of 0.95, and specificity of 0.79 to differentiate sPE and non-PE pregnant women, using CBC parameters of neutrophils count, mean corpuscular hemoglobin (MCH), and the aggregate index of systemic inflammation (AISI). In addition, we provided a ridge regression equation using the same three CBC parameters, which is fully interpretable and achieved an AUROC of 0.79±0.10 to differentiate the both groups. Moreover, we also showed that a monocyte count lower than 490/mm3\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$490 /mm^{3}$$\\end{document} yielded a sensitivity of 0.71 and specificity of 0.72.ConclusionOur study showed that ML-powered CBC could be used as a biomarker for sPE diagnosis support. In addition, we showed that a low monocyte count alone could be an indicator of sPE.SignificanceAlthough preeclampsia has been extensively studied, no laboratory biomarker with favorable cost-effectiveness has been proposed. Using artificial intelligence, we proposed to use the CBC, a low-cost, fast, and well-spread blood test, as a biomarker for sPE.

The transcriptome of CD14 + CD163 - HLA-DR low monocytes predicts mortality in Idiopathic Pulmonary Fibrosis.

The association between immune-cell-specific transcriptomic profiles and Idiopathic Pulmonary Fibrosis (IPF) mortality is unknown. To determine immune-cell-specific transcriptomic profiles associated with IPF mortality. We profiled peripheral blood mononuclear cells (PBMC) in 18 participants [University of South Florida: IPF, COVID-19, post-COVID-19 Interstitial Lung Disease (Post-COVID-19 ILD), controls] by single-cell RNA sequencing (scRNA-seq) and identified 16 immune-cell-specific transcriptomic profiles. The Scoring Algorithm of Molecular Subphenotypes (SAMS) was used to calculate Up-scores based on these 16 gene profiles. Their association with outcomes was investigated in peripheral blood, Bronchoalveolar Lavage (BAL) and lung tissue of N=416 IPF patients from six cohorts. Findings were validated in an independent IPF, PBMC scRNA-seq dataset (N=38). Cox-regression models demonstrated that 230 genes from CD14 + CD163 - HLA-DR low circulating monocytes predicted IPF mortality [Pittsburgh (p=0.02), Chicago (p=0.003)]. PBMC proportions of CD14 + CD163 - HLA-DR low monocytes were higher in progressive versus stable IPF (Yale, 0.13±0.05 versus 0.09±0.05, p=0.034). Receiving operating characteristic identified a 230 gene, Up-score >41.84 (Pittsburgh) predictive of mortality in Chicago (HR: 6.58, 95%CI: 2.15-20.13, p=0.001) and in pooled analysis of BAL cohorts (HR: 2.20, 95%CI: 1.44-3.37, p=0.0003). High-risk patients had decreased expression of the T-cell co-stimulatory genes CD28 , ICOS , ITK and LCK (Pittsburgh and Chicago, p<0.01). 230 gene-up-scores negatively correlated with Forced Vital Capacity (FVC) in IPF lung tissues (LGRC, rho=-0.2, p=0.02). Results were replicated using a subset of 13 genes from the 230-gene signature (pooled PBMC cohorts - HR: 5.34, 95%CI: 2.83-10.06, p<0.0001). The transcriptome of CD14 + CD163 - HLA-DR low monocytes is associated with increased IPF mortality.

Oral Complications Due to Medication in Stevens-Johnson Syndrome Patient with Systemic Involvement

Background: Stevens-Johnson Syndrome (SJS) is a drug-induced hypersensitivity reaction involving mucocutaneous with various trigger factors including drugs and herpes simplex virus. Objective: This case report aimed to discuss oral complications due to medication in SJS patients with systemic disease involvement. Case: A 51-year-old man was referred to the Department of Oral Medicine, Faculty of Dentistry, Padjadjaran University, at Hasan Sadikin Hospital from the dermatology and venereology department complaining of pain in the oral cavity, especially when eating and drinking two months before, with a history of phenytoin, salbutamol and theophylline therapy. Extraoral examination showed erosive lesions and tended to bleed serosanguinolenta crusts on the lips. Intraoral there were erosive lesions and white plaque on the tongue, buccal mucosa, labial mucosa, and palate, as well as dental caries and calculus. Blood examination showed low hemoglobin, hematocrit, erythrocytes, lymphocytes, monocytes, SGOT, and sodium levels, while HbA1c, random, fasting, and 2 HPP glucose levels were high. Reactive Anti-HSV-1 IgG and rheumatoid factor. KOH examination showed positive spores, hyphae, pseudohypha, and budding cells. The diagnosis was SJS-associated oral lesions with HSV-1 virus infection, oral candidiasis accompanied with diabetes mellitus. For diabetes mellitus treatment, he was referred to the internal medicine department. Case Management: The therapy was 0.9% NaCl for lip compress, acyclovir tablet, nystatin oral suspension, chlorhexidine digluconate 0,12% mouthwash, folic acid, and vitamin B12. Oral lesions were improved significantly after diabetes mellitus was treated. Conclusion: The SJS patient was susceptible to complications in the oral cavity, especially fungal and viral infections due to the received medication.

Autopsy findings from patients diagnosed with COVID-19 demonstrate unique morphological patterns in bone marrow and lymph node

AimsThe identification of haemophagocytosis in bone marrow (BM) is recurrently identified in patients with severe COVID-19. These initial COVID-19 autopsy studies have afforded valuable insight into the pathophysiology of this...

Chronic myelomonocytic leukemia with ring sideroblasts/SF3B1 mutation presents with low monocyte count and resembles myelodysplastic syndromes with-RS/SF3B1 mutation in terms of phenotype and prognosis.

Chronic myelomonocytic leukemia (CMML) and myelodysplastic syndromes (MDS) with ring sideroblasts (RS) or SF3B1 mutation (MDS-RS/SF3B1) differ in many clinical features, but share others, such as anemia. RS and SF3B1 mutation can also be found in CMML. We compared CMML with and without RS/SF3B1 and MDS-RS/SF3B1 considering the criteria established by the 2022 World Health Organization classification. A total of 815 patients were included (CMML, n=319, CMML-RS/SF3B1, n=172 and MDS-RS/SF3B1, n=324). The percentage of RS was ≥15% in almost all CMML-RS/SF3B1 patients (169, 98.3%) and most (125, 72.7%) showed peripheral blood monocyte counts between 0.5 and 0.9 x109/L and low risk prognostic categories. CMML-RS/SF3B1 differed significantly from classical CMML in the main clinical characteristics, whereas it resembled MDS-RS/SF3B1. At a molecular level, CMML and CMML-RS/SF3B1 had a significantly higher frequency of mutations in TET2 (mostly multi-hit) and ASXL1 (p=0.013) and CMML had a significantly lower frequency of DNMT3A and SF3B1 mutations compared to CMML/MDS-RS/SF3B1. Differences in the median overall survival among the three groups were statistically significant: 6.75 years (95% confidence interval [CI] 5.41-8.09) for CMML-RS/SF3B1 vs. 3.17 years (95% CI 2.56-3.79) for CMML vs. 16.47 years (NA) for MDS-RS/SF3B1, p<0.001. Regarding patients with CMML and MDS, both with SF3B1 mutation, survival did not significantly differ. CMML had a higher risk of transformation to acute myeloid leukemia (24% at 8 years, 95%CI 19%-30%). CMML-RS/SF3B1 mutation resembles MDS-RS/SF3B1 in terms of phenotype and clearly differs from CMML. The presence of ≥15% RS and/or SF3B1 in CMML is associated with a low monocyte count. SF3B1 mutation clearly improves the prognosis of CMML.

The association between systemic immune-inflammation index and cardiotoxicity related to 5-Fluorouracil in colorectal cancer

Background and aimsThe cardiotoxicity related to 5-Fluorouracil (5-FU) in cancer patients has garnered widespread attention. The systemic immune-inflammation index (SII) has recently been identified as a novel predictive marker for the development of cardiovascular illnesses in individuals without pre-existing health conditions. However, it remains unclear whether the levels of SII are linked to cardiotoxicity related to 5-FU. This retrospective study aims to fill this knowledge gap by examining the correlation between SII and cardiotoxicity related to 5-FU in a colorectal cancer cohort.MethodsThe study comprised colorectal cancer patients who received 5-FU-based chemotherapy at the affiliated cancer hospital of Guizhou Medical University between January 1, 2018 and December 31, 2020. After adjustment for confounders and stratification by tertiles of the interactive factor, linear regression analyses, curve fitting and threshold effect analyses were conducted.ResultsOf the 754 patients included final analysis, approximately 21% (n = 156) of them ultimately experienced cardiotoxicity related to 5-FU. Monocytes (M) was found as an influential element in the interaction between SII and cardiotoxicity related to 5-FU. In the low tertile of M (T1: M ≤ 0.38 × 109/L), increasing log SII was positively correlated with cardiotoxicity related to 5-FU (Odds Ratio [OR], 8.04; 95% confidence interval [95%CI], 1.68 to 38.56). However, a curvilinear relationship between log SII and cardiotoxicity was observed in the middle tertile of M (T2: 0.38 < M ≤ 0.52 × 109/L). An increase in log SII above 1.37 was shown to be associated with a decreased risk of cardiotoxicity (OR, 0.14; 95%CI, 0.02 to 0.88), indicating a threshold effect. In the high tertile of M (T3: M > 0.52 × 109/L), there was a tendency towards a negative linear correlation between the log SII and cardiotoxicity was observed (OR, 0.85; 95%CI, 0.37 to 1.98).ConclusionOur findings suggest that SII may serve as a potential biomarker for predicting cardiotoxicity related to 5-FU in colorectal cancer patients. SII is an independent risk factor for cardiotoxicity related to 5-FU with low monocytes levels (T1). Conversely, in the middle monocytes levels (T2), SII is a protective factor for cardiotoxicity related to 5-FU but with a threshold effect.

Immunomodulatory Synthetic Glycocluster Molecule Prevents Melanoma Growth In Vivo.

Triacedimannose (TADM) is a synthetic trivalent acetylated glycocluster and a transmembrane macrophage activator independent of the mannose receptor. TADM induces Th1-type immune responses and suppresses Th2-type cytokines in acute and chronic allergic inflammation models in vivo. We, therefore, wanted to test whether TADM could also facilitate anti-tumour tissue responses similar to what has been observed for the immune checkpoint inhibitors, such as anti-PD-1 and anti-CTLA-4. A syngeneic mouse melanoma model was selected since metastatic melanoma has been successfully targeted by checkpoint inhibitors in the clinic. TADM inhibited the growth of B16 mouse melanoma tumours at levels comparable to an anti-PD-1 antibody. TADM-treated tumours encompassed significantly more apoptotic cells as measured by TUNEL staining, and interferon-gamma (IFN-γ) expression was increased in the spleens of TADM-treated mice compared to untreated controls. TADM-treated mice also demonstrated increased Ly6 C low monocytes and neutrophils in the spleens. However, TADM-treated tumours showed no discernible differences in infiltrating immune cells. TADM can alone suppress the growth of melanoma tumours. TADM likely activates M1 type macrophages, type N1 neutrophils, and CD8+ and Th1 T cells, suppressing the type 2 immune response milieu of melanoma tumour with a strong type 1 immune response.

Abstract 3054: Proinflammatory Phenotypes And Role Of Ly-6c Low Monocytes In Atherosclerosis With Hypertriglyceridemia

Background: Monocytes play pivotal roles in atherosclerosis. Ly-6C low monocytes, also known as patrolling monocytes, become foamy in the circulation of mice with hyperlipidemia. However, the role of Ly-6C low monocytes in atherosclerosis with hypertriglyceridemia (HTG) remains to be determined. Aim: To examine the phenotypes and role of Ly-6C low monocytes in atherosclerosis in HTG. Method: Ldlr -/- ApoC3Tg (transgenic expression of human ApoC3) mice fed a western high-fat diet (WD) were used to examine the effects of HTG on monocyte lipid accumulation (foamy monocyte formation), phenotypes, and contributions to atherosclerosis. Results: Plasma triglycerides in Ldlr -/- ApoC3Tg mice were 5.1 times higher than in Ldlr -/- controls at 6 weeks on WD, correlating with more than twice the increase in aortic atherosclerosis (P&lt;0.01). Consistently, Ly-6C low compared to Ly-6C high monocytes in Ldlr -/- mice had increased lipid accumulation and exhibited elevated levels of TNFα, IL-6, IL-1β, Plin2, and PPAR-γ. In Ldlr -/- ApoC3Tg mice, Ly-6C low (also CD11c + ) monocytes demonstrated even greater lipid accumulation and adhesion to VCAM-1 and had further elevations in levels of TNFα, IL-6, IL-1β, Plin2, and PPAR-γ than CD11c + (Ly-6C low ) monocytes in Ldlr -/- controls, indicating an enhanced inflammatory response and lipid accumulation under HTG. Furthermore, compared to Ldlr -/- control mice, Ldlr -/- ApoC3Tg mice had an increase in the level of SIRPα (a ligand of the “don’t-eat me” signal CD47) on Ly-6C low monocytes, indicative of impairment in efferocytosis and clearance of apoptotic cells. Additionally, nuclear c-Rel mRNA and protein levels were upregulated in Ly-6C low monocytes of Ldlr -/- ApoC3Tg mice compared to Ldlr -/- controls, suggesting activation of the NF-κB pathway in Ly-6C low monocytes by HTG. Finally, depletion of Ly-6C low monocytes reduced atherosclerosis by 45% (P&lt;0.05) in Ldlr -/- ApoC3Tg mice, indicating a role of Ly-6C low monocytes in atherosclerosis in HTG. Conclusions: HTG accelerates atherogenesis in Ldlr -/- mice, possibly by increasing lipid accumulation and inflammatory phenotypic changes in monocytes, primarily CD11c + Ly-6C low monocytes.

Intestinal stroma guides monocyte differentiation to macrophages through GM-CSF

Stromal cells support epithelial cell and immune cell homeostasis and play an important role in inflammatory bowel disease (IBD) pathogenesis. Here, we quantify the stromal response to inflammation in pediatric IBD and reveal subset-specific inflammatory responses across colon segments and intestinal layers. Using data from a murine dynamic gut injury model and human ex vivo transcriptomic, protein and spatial analyses, we report that PDGFRA+CD142−/low fibroblasts and monocytes/macrophages co-localize in the intestine. In primary human fibroblast-monocyte co-cultures, intestinal PDGFRA+CD142−/low fibroblasts foster monocyte transition to CCR2+CD206+ macrophages through granulocyte-macrophage colony-stimulating factor (GM-CSF). Monocyte-derived CCR2+CD206+ cells from co-cultures have a phenotype similar to intestinal CCR2+CD206+ macrophages from newly diagnosed pediatric IBD patients, with high levels of PD-L1 and low levels of GM-CSF receptor. The study describes subset-specific changes in stromal responses to inflammation and suggests that the intestinal stroma guides intestinal macrophage differentiation.

Along with PaO2/FiO2 ratio and lymphopenia, low HLA-DR monocytes are the only additional parameter that independently predicts the clinical course of undifferentiated SARS-CoV-2 patients in emergency departments.

Because one-third of patients deteriorate after their admission to the emergency department, assessing the prognosis of COVID-19 patients is of great importance. However, to date, only lymphopenia and the partial pressure of oxygen/fraction of inspired oxygen (PaO2/FiO2) ratio have been reported as partly predictive of COVID-19-related further deterioration, and their association has not been evaluated. We asked whether other key biomarkers of SARS-CoV-2 immunologic defects-increase in circulating immature granulocytes, loss of monocyte HLA-DR (mHLA-DR) expression, and monocyte differentiation blockade-could also predict further COVID-19 deterioration. A series of 284 consecutive COVID-19 patients, with the sole inclusion criterion of being an adult, were prospectively enrolled at emergency department admission (day 0) of 2 different hospitals: 1 for the exploratory cohort (180 patients) and 1 for the confirmatory cohort (104 patients). Deterioration was assessed over the next 7 days. Neither increased immature granulocyte levels nor monocyte differentiation blockade predicted patient worsening. Among more than 30 clinical, biological, and radiological parameters, the value of decreased P/F ratio and lymphopenia for prediction of further COVID-19 deterioration was strongly confirmed, and the loss of mHLA-DR was the only additional independent marker. Combined together in a simple OxyLymphoMono score, the 3 variables perfectly predicted patients who did not worsen and correctly predicted worsening in 59% of cases. By highlighting lymphocyte and monocyte defects as preceding COVID-19 deterioration, these results point on early immunosuppression in COVID-19 deterioration. Combining P/F ratio, lymphopenia, and loss of mHLA-DR together in a simple and robust score could offer a pragmatic method for COVID-19 patient stratification.

Abstract WP317: Phagocytic Potentials of Purinergic Receptor P2x4 Blockade After Ischemic Stroke

Backgrounds: We previously showed that genetic deletion as well as short term pharmacological inhibition of P2X4R, a purinergic receptor for adenosine triphosphate ATP, provides neuroprotection by reducing acute excessive inflammation and thus can be potential drug targets to treat ischemic stroke. However, the mechanism how P2X4R blockade provide long term benefits are not known. Here we hypothesize that P2X4R blockade increase phagocytic uptake of damaged or dead tissue and facilitate swift recovery. Methods: We used both 1μm size fluorescent phagocytic bead (Polyscience Inc) and pHrodo Red zymosan A Bioparticles conjugate (Thermofisher) to study in vitro and ex-vivo phagocytic uptake by LPS (200 μg/ml for 2 hours) primed Bone Marrow Derived Macrophages (BMDMs) from P2X4R KO and littermate WT mice. We also used flow sorted Ly6C hi and low monocytes for phagocytic uptake study obtained from perilesional ipsilateral mouse brain tissue of P2X4R KO and littermate WT mice after 3 or 7 days of stroke. The later mice were subjected to 60 mins of transient middle cerebral artery occlusion. Results: Both phagocytic beads and pHrodo bioparticle uptake study suggest significantly higher uptake (1.5 and 2-fold P&lt;0.05 vs WT student t test n=4 mice/group x 2 technical replication) by BMDMs obtained from P2X4R KO mice. FACS sorted infiltrated Ly6Chi inflammatory monocytes from global P2X4R KO mice brain after 3 days of stroke show significantly (*P&gt;0.05 vs WT) high phagocytic bead uptake as compared to WT control. Further these monocytes also showed elevated expression of CD36, a marker for scavenger receptor at 7 days after stroke. Conclusions: This study suggest that P2X4R blockade provide swift and sustained neuroprotection by increasing phagocytic uptake of damage tissue after ischemic stroke.

Molecular Classification of Chronic Myelomonocytic Leukemia: Results of the Analysis of an International Cohort of 2,471 Patients

Background. Chronic myelomonocytic leukemia (CMML) is a rare myeloid neoplasm that shares both dysplastic and proliferative features and includes patients with highly heterogeneous clinical manifestations and prognosis. Despite known co-mutation patterns that are suggesting of a CMML diagnosis such as TET2 and SRSF2 mutations, the current diagnosis and classification schemes rely solely on morphological criteria that fail to capture the genomic heterogeneity of the disease. Here we perform an unsupervised analysis using only genomic features to generate a novel classification schema of CMML patients with unique clinical features. Methods. A cohort of 2,471 CMML patients defined according to WHO 2016 criteria from European and US centers was analyzed. Bayesian network analysis and hierarchical Dirichlet processes were used to identify genomic associations and subgroups as a basis to define a molecular classification of CMML. Bayesian networks allow to infer the structure of conditional dependencies among mutations. Dirichlet processes were applied to define clusters capturing broad dependencies among all gene mutations and cytogenetic abnormalities. Multivariate logistic regression analysis was applied to compare clinical and hematological characteristics among different groups. ( JCO 2021 39:11, 1223-1233) Results. Baseline clinical and molecular characteristics are summarized in Table 1. We identified 11 clusters defined according to specific genomic features (Table 2). Two clusters were primarily defined by the presence of mutations in splicing genes SRSF2 and ZRSR2, frequently associated with TET2 mutations. Patients assigned to these clusters showed myelodysplastic features according to French American British (FAB) criteria, had a low rate of peripheral blood cytopenias, and low blast counts. Additionally, two clusters were characterized by the same mutations in splicing genes (i.e., SRSF2 or ZRSR2 with associated TET2), together with co-mutations in genes that correlate with a more aggressive phenotype ( RUNX1, ASXL1, EZH2). These groups showed overlapping myelodysplastic and myeloproliferative features by FAB, intermediate-risk per prognostic scores, and decreased survival as compared with the first 2 clusters. A distinct cluster was characterized by SF3B1 gene mutations (frequently associated with TET2 or DNMT3A). The SF3B1 cluster was marked by a dysplastic phenotype and more severe anemia (with a high rate of transfusion-dependence) with respect to the other genomic groups. Four clusters (27.8% of the overall population) were characterized by the presence of mutations in signal transduction and tyrosine kinase pathways genes ( NRAS/KRAS, CBL, JAK2 and SETBP1 often associated with ASXL1/TET2, respectively). Patients within these groups showed a mostly proliferative phenotype and intermediate survival. The presence of JAK2 mutations appeared to confer a better prognosis compared with other proliferative clusters. A minority of patients (4.6%) were assigned to two high-risk clusters, defined by TP53 mutations (often biallelic) with complex karyotype, and AML-like mutations ( NPM1, IDH1/2, FLT3), respectively. Patients in these clusters were predominantly dysplastic by FAB, had a high prevalence of severe cytopenias, higher blast counts, and a dismal outcome. Finally, we identified a cluster of patients defined by the absence of gene mutations and chromosomal abnormalities (4.5%). The majority of patients assigned to this cluster were characterized by a dysplastic phenotype, had low absolute monocyte counts, and favorable outcome. A subset of patients (18.9%) could not be assigned to a specified cluster in this preliminary analysis. An independent validation of this molecular classification is currently being conducted on a US prospective cohort (n=650). Conclusion. Our results provide proof of concept for a molecular classification of CMML based on the identification of homogeneous genomic signatures and associated clinical features. 15% of patients showed clear overlapping features with other myeloid neoplasms, thus providing the rational to refine the boundaries among different clinical entities. A molecular classification of CMML may provide a basis to define a next-generation prognostic tool and improve clinical decision-making.

A Novel Model Combining Circulating Absolute Monocyte Counts and a Four-Gene Monocyte Signature in Leukapheresis Identifies Lymphoma Patients at Very High Risk of Progression after Tisagenlecleucel or Axicabtagene Ciloleucel Therapy

Introduction CD19-directed chimeric antigen receptor (CAR) T can induce durable remissions in a fraction of relapsed/refractory large B-cell lymphomas (R/R LBCL), but 60% of patients (pts) still relapse. Biological mechanisms explaining lack of responses are emerging but they are largely unsuccessful in predicting disease response at the patient level. Aim To address the potential role of lymphocyte features before CAR T manufacturing, we performed transcriptomic and functional evaluations of leukapheresis products (LK) in 95 R/R LBCL pts enrolled in a prospective observational study, to identify correlates of response and survival to tisagenlecleucel (Tisa-cel) and axicabtagene ciloleucel (Axi-cel). Methods RNA of sorted CD3+ cells from LK was digitally quantified using the nCounter CAR T Characterization Panel (NanoString). Flow cytometry (FCM) was used for CAR T enumeration and monocyte analysis. Disease response at day 90 was assessed by computed tomography (CT) and positron emission tomography-CT (PET/CT) according to Lugano criteria. Statistical analyses were performed by GraphPad Prism v.9.00 and R v.4.1.2. Results We identified a 4-gene transcriptional signature (including MS4A4A, CD86, CD163 and SIGLEC5) able to divide LBCL pts into two groups, [namely Expressors (EXP) and poor-Expressors (poor-EXP)], characterized by significantly different survival probabilities. EXP have shorter PFS (P&amp;lt;0.0001: median PFS for EXP was 61 days and not reached for poor-EXP) (Fig. 1) and are less likely to respond (P&amp;lt;0.01). As the identified genes are highly expressed in monocytes but not in T cells, we sought to define whether monocytes contained in LK have a role in determining post CAR T outcome. In line with the fact that EXP had higher frequencies of monocytes in LK by FCM, we confirmed elevated absolute monocyte counts (AMC) both in LK (P&amp;lt;0.01) and in peripheral blood (PB) (P&amp;lt;0.05) of EXP. We then checked if monocyte levels could influence response and survival. When compared to responders, non-responders (NR) had higher LK AMC (P&amp;lt;0.01), higher percentages of monocytes (P&amp;lt;0.01) but also decreased percentages of lymphocytes (P&amp;lt;0.001), a parameter that was also associated with reduced PFS [HR: 0.97 (CI 0.96 - 0.99) P=0.01]. Consistent with the fact that circulating monocytes significantly correlated with those in LK (r= 0.6621, P&amp;lt;0.0001), NR also had significantly higher AMC (P&amp;lt;0.001) and AMC levels in PB negatively correlated with PFS [HR: 1.8 (CI 1.2 - 2.9) P=0.01]. Additionally, pts with serum C-reactive protein (CRP) and ferritin levels higher than the upper normal limit, displayed significantly higher AMC (P&amp;lt;0.001 for CRP; P&amp;lt;0.05 for ferritin). Accordingly, univariate analysis indicated that high ferritin, CRP and LDH levels at the time of LK were associated with the lack of response, but only high serum ferritin levels correlated with shorter PFS [HR: 2.86 (CI 1.25-6.55) P=0.013]. We next investigated if the combination of the monocyte signature and high ferritin levels and/or high circulating monocytes could better identify a group of pts at high risk of progression, compared to the expression of the 4 genes alone. Remarkably, albeit high ferritin levels and monocyte counts are overall associated with significantly reduced survival, the shortest PFS was achieved in EXP with monocyte counts above the median (P=0.0234: median PFS for EXP with high monocytes was 32.5 days and for EXP with low monocytes 209 days) (Fig. 2), thus supporting the hypothesis that CAR T treatment outcome is influenced by the presence of elevated levels of monocytes expressing the 4 genes. Concatenated monocyte FCM data from 30 pts analyzed with FlowSOM, followed by a consensus clustering algorithm combined with t-SNE (t-stochastic neighbor embedding), indicated that EXP had higher frequencies of CD14hi monocytes co-expressing MS4A4A, CD86, CD163 and SIGLEC5 at the protein level than poor-EXP (P&amp;lt;0.05), and accordingly, the presence of this population was also associated with lack of response (P&amp;lt;0.05). Conclusions Our data suggest that the poor outcome of some LBCL pts receiving CAR T is due to the presence of high monocytes levels expressing a four-gene signature in an inflammatory microenvironment. Additionally, the pre-manufacturing combined analysis of the gene signature and of PB AMC, could be used to plan trials evaluating CAR T cells versus other newly approved therapies such as bispecific antibodies.

Serum Interferon Gamma (IFN-ɣ) Levels and Hematological Indices in Patients with HIV-MTB Co-Infection in North-Eastern Nigeria

Introduction: The dual epidemic of human immunodeficiency virus (HIV) infection and Mycobacterium tuberculosis (MTB) poses significant health challenges, particularly in sub-Saharan Africa. Understanding the immune response and hematological changes in HIV-MTB co-infection is crucial for better management of affected individuals. Objectives: This study aimed to evaluate the serum levels of IFN-ɣ and hematological indices in patients with HIV-MTB co-infection in North-Eastern Nigeria, as well as explore any potential relationships between these factors. Methods: A total of 88 participants were enrolled in the study, including 44 antiretroviral therapy-naive patients with HIV-MTB co-infection (study group) and 44 HIV mono-infected individuals as controls. Data on personal biodata and clinical details were collected using an interviewer-administered questionnaire. Blood samples were obtained from each participant and analyzed for IFN-ɣ levels using ELISA and hematological indices using an automated hematology analyzer. Statistical analysis, including Mann-Whitney U test, independent samples t-test, and Pearson's correlation analysis, was conducted to compare the study and control groups and assess the relationship between IFN-ɣ levels and hematological parameters. Results: Serum IFN-ɣ was insignificantly increased in the study group compared to the control group (p=0.093). The WBC count was also significantly reduced in the study group compared to the control group (p=0.038). The HGB, HCT, MCV and MCH were significantly reduced in the study group compared to the control group (p=0.001, 0.001, 0.002 and 0.001) respectively. Participants with HIV-TB co-infection have insignificantly increased serum IFN-ɣ levels, low total WBC, lymphocyte and monocyte counts compared to those with HIV mono-infection. In conclusion, participants with HIV-TB co-infection have insignificant increased serum IFN-ɣ levels, low total WBC, and lymphocyte and monocyte counts compared to those with HIV mono-infection. There was no correlation of IFN-ɣ with any of the haematological indices.

The Prognostic Role of Preoperative Hematological and Inflammatory Indices in Canine Appendicular Osteosarcoma.

Hematological indices play a prognostic role in human osteosarcoma (OSA), but data are limited in dogs. The aim of this retrospective multicentric cohort study was to investigate the prognostic significance of pre-operative hematological/inflammatory indices in a cohort of client-owned dogs with appendicular OSA receiving standardized treatment. Cut-offs associated with progression-free survival (PFS) for pre-operative hematological values/ratios were established using the minimal p-value approach. Historical prognostic factors were also assessed. Statistical analyses were performed for the whole population and after the exclusion of sighthounds. Fifty-nine dogs were included (13 were sighthounds). Multivariable analysis revealed that a low neutrophil count (<4.37 × 109/L, HR0.28, CI 95% 0.13-0.61, p = 0.001), a high red blood cell count (≥7.91, HR3.5, CI 95% 1.56-7.9, p = 0.002), and a proximal humerus location (HR3.0, CI 95% 1.48-6.1, p = 0.002) were associated with shorter PFS. In the sighthound-only population, only OSA location was significantly associated with PFS in univariable analysis. When sighthounds were excluded, a low neutrophil count, a low monocyte count, and a proximal humerus location were associated with shorter PFS, in multivariable analysis. Neutrophil count and possibly monocyte and red blood cell counts can be useful prognostic markers in canine OSA treated with amputation and adjuvant carboplatin. However, not all indices are appropriate in sighthounds.

Monocytic HLA-DR expression in type 2 diabetes mellitus: Impact on disease susceptibility

Monocytes are innate immune cells that act as antigen-presenting cells. HLA-DR is a cell surface protein that plays a crucial role in the immune system by presenting antigens to CD4 T Cells (T helper). Based on surface expression, monocytes can differentiate into three subsets; classical monocytes (CD14+CD16-), intermediates monocytes (CD14+ CD16+), and non-classical monocytes (CD14dim CD16+). Among these subsets, classical monocytes have the lowest, and intermediate monocytes have the highest expression of HLA-DR. Monocytes are very sensitive to environmental changes, like hyperglycemia in Type 2 Diabetes Mellitus (T2DM). Metabolic changes in T2DM conditions trigger changes in immune cells including monocytes. Chronic low-grade inflammation in T2DM decreased HLA-DR expression in all monocyte subsets. Monocytes that do not express or lose HLA-DR expression are known as CD14+HLA-DR-/low monocyte and is immunosuppressive. This increase in monocytes in T2DM conditions is related to the ability of monocytes as antigen-presenting cells. Decreased HLA-DR expression increased the risk of severity and susceptibility to several diseases such as COVID-19 infection, TB, cancer, or sepsis. The alterations in HLA-DR expression are one of the factors that make T2DM a comorbid disease. This literature review aims to explore monocytic HLA-DR in T2DM conditions hence increasing the risk of severity and susceptibility to disease. This literature is expected to be the basis for research on the potential of HLA-DR as a prevention or treatment for T2DM patients.

Vpx requires active cellular dNTP biosynthesis to effectively counteract the anti-lentivirus activity of SAMHD1 in macrophages

HIV-1 replication in primary monocyte-derived macrophages (MDMs) is kinetically restricted at the reverse transcription step due to the low deoxynucleoside triphosphates (dNTP) pools established by host dNTPase, SAM and HD domain containing protein 1 (SAMHD1). Lentiviruses such as HIV-2 and some Simian immunodeficiency virus counteract this restriction using viral protein X (Vpx), which proteosomally degrades SAMHD1 and elevates intracellular dNTP pools. However, how dNTP pools increase after Vpx degrades SAMHD1 in nondividing MDMs where no active dNTP biosynthesis is expected to exists remains unclear. In this study, we monitored known dNTP biosynthesis machinery during primary human monocyte differentiation to MDMs and unexpectedly found MDMs actively express dNTP biosynthesis enzymes such as ribonucleotide reductase, thymidine kinase 1, and nucleoside-diphosphate kinase. During differentiation from monocytes the expression levels of several biosynthesis enzymes are upregulated, while there is an increase in inactivating SAMHD1 phosphorylation. Correspondingly, we observed significantly lower levels of dNTPs in monocytes compared to MDMs. Without dNTP biosynthesis availability, Vpx failed to elevate dNTPs in monocytes, despite SAMHD1 degradation. These extremely low monocyte dNTP concentrations, which cannot be elevated by Vpx, impaired HIV-1 reverse transcription in a biochemical simulation. Furthermore, Vpx failed to rescue the transduction efficiency of a HIV-1 GFP vector in monocytes. Collectively, these data suggest that MDMs harbor active dNTP biosynthesis and Vpx requires this dNTP biosynthesis to elevate dNTP levels to effectively counteract SAMHD1 and relieve the kinetic block to HIV-1 reverse transcription in MDMs.

Abstract 107: CaMK4 is a Novel Regulator of Myeloid Phenotype and Function in Atherosclerosis

Background: Chronic inflammation is a major driver of atherosclerotic cardiovascular disease, and therapeutics that target inflammation reduce clinical cardiac events beyond levels seen with conventional strategies targeting cholesterol alone. Recent work by our group and others has demonstrated that advanced atherosclerosis is also characterized by the failure of an active repair process termed ‘inflammation resolution’. Strategies that boost resolution and break the cycle of chronic inflammation promotes plaque stability. Hypothesis: Our review of publicly available RNAseq data revealed an increase in expression of Calcium/calmodulin-dependent protein kinase 4 (CaMK4) in advanced/unstable regions of plaque within human carotid arteries as well as in myeloid cells derived from atherosclerotic murine aortas. Therefore, we hypothesized that CaMK4 promotes inflammation and impairs resolution. Results and Methods: Control and Camk4 -/- mice were injected with AAV-8 PCSK9 virus and fed a high-fat high-cholesterol Western diet for 12 weeks. Cross-sectional analysis of aortic roots showed that Camk4 -/- mice had significantly less plaque burden than Controls. Flow cytometric analysis revealed elevated monocyte numbers in Camk4 -/- mice compared to Control mice. Further analysis demonstrated increased skewing of Camk4 -/- monocyte populations toward a Ly6c low subset, indicating a more pro-reparative phenotype compared to Control mice. We considered whether CaMK4 may regulate a gene signature driving monocyte conversion and found that Camk4 -/- monocytes expressed higher levels of Nr4a1, Cebpb, and Klf2, which have been shown to promote conversion to Ly6c low monocytes. As Ly6c low monocytes give rise to pro-reparative macrophages, we generated bone marrow-derived macrophages and found that loss of CaMK4 in vitro led to higher levels of pro-reparative cytokines being produced in response to LPS stimulation as well as more efficient clearance of apoptotic cells when compared to Controls. Conclusions: Overall, these findings suggest that CaMK4 regulates myeloid phenotype in vitro and in vivo and that loss of CaMK4 promotes pro-resolving macrophage function. Therefore, targeting CaMK4 may offer a unique way to target atheroprogression.

Are preoperative monocytes and HDL values ​​an early predictor of recurrence in the surgical ablation treatment of atrial fibrillation?

Objective: Monocyte HDL (high-density lipoprotein) cholesterol ratio has been accepted as a newly emerging cardiovascular prognostic marker. The aim of this study is to investigate the determinants of monocyte and HDL cholesterol values ​​in the early recurrence of atrial fibrillation (AF) treated with cryoablation and radiofrequency ablation.&#x0D; &#x0D; Materials and Methods: This retrospective study was conducted between September 2006 and July 2014 in the Department of Cardiovascular Surgery, Adana City Hospital, Health Sciences University, including 100 patients who underwent surgical AF ablation with open heart surgery. Logistic regression analysis was used to determine monocytes and HDL cholesterol values, which are among the factors affecting recurrence in the first 3 months postoperatively.&#x0D; &#x0D; Results: 100 patients who underwent surgical ablation together with open heart surgery were evaluated for early postoperative recurrence. According to the logistic regression analysis, the most effective features and measurements for early recurrence were diabetes mellitus (DM), AF duration, left atrial diameter, and low HDL cholesterol and high monocyte values ​​before the procedure.&#x0D; &#x0D; Conclusion: Preoperative low HDL and high monocyte values ​​can be considered as a determining factor for early recurrence in surgical ablation treatment of AF.

Immune Profiling Panel Gene Set Identifies Critically Ill Patients With Low Monocyte Human Leukocyte Antigen-DR Expression: Preliminary Results From the REAnimation Low Immune Status Marker (REALISM) Study.

There is a crucial unmet need for biomarker-guided diagnostic and prognostic enrichment in clinical trials evaluating immune modulating therapies in critically ill patients. Low monocyte expression of human leukocyte antigen-DR (mHLA-DR), considered as a reference surrogate to identify immunosuppressed patients, has been proposed for patient stratification in immunostimulation approaches. However, its widespread use in clinic has been somewhat hampered by technical constraints inherent to flow cytometry technology. The objective of the present study was to evaluate the ability of a prototype multiplex polymerase chain reaction tool (immune profiling panel [IPP]) to identify immunosuppressed ICU patients characterized by a low mHLA-DR expression. Retrospective observational cohort study. Adult ICU in a University Hospital, Lyon, France. Critically ill patients with various etiologies enrolled in the REAnimation Low Immune Status Marker study (NCT02638779). None. mHLA-DR and IPP data were obtained from 1,731 blood samples collected from critically ill patients with various etiologies and healthy volunteers. A partial least square regression model combining the expression levels of IPP markers was trained and used for the identification of samples from patients presenting with evidence of immunosuppression, defined here as mHLADR less than 8,000 antibodies bound per cell (AB/C). The IPP gene set had an area under the receiver operating characteristic curve (AUC) of 0.86 (95% CI 0.83-0.89) for the identification of immunosuppressed patients. In addition, when applied to the 123 patients still in the ICU at days 5-7 after admission, IPP similarly enriched the number of patients with ICU-acquired infections in the immunosuppressed group (26%), in comparison with low mHLA-DR (22%). This study reports on the potential of the IPP gene set to identify ICU patients presenting with mHLA-DR less than 8,000 AB/C. Upon further optimization and validation, this molecular tool may help in the stratification of patients that could benefit from immunostimulation in the context of personalized medicine.