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While the main role of phagocytic scavenger cells consists of the neutralization and elimination of pathogens, they also keep the body fluids clean by taking up and breaking down waste material. Since a build-up of waste is thought to contribute to the aging process, these cells become particularly pertinent in the research field of aging. Nevertheless, a direct link between their scavenging functions and the aging process has yet to be established. Integrative approaches involving various model organisms hold promise to elucidate this potential, but are lagging behind since the diversity and evolutionary relationship of these cells across animal species remain unclear. In this perspective, we review the current knowledge associating phagocytic scavenger cells with aging in vertebrate and invertebrate animals, as well as put forward important questions for further exploration. Additionally, we highlight future challenges and propose a constructive approach for tackling them. [ABSTRACT FROM AUTHOR]

Background/aim: The transfusion of blood products is a life-saving clinical practice in patients with bleeding, hemoglobinopathy, and cancer. It was aimed herein to analyze the frequency and types of blood component-related acute transfusion reactions (ATRs) in pediatric patients. Materials and methods: This retrospective study was conducted at a tertiary care academic pediatric hospital. Results: During the study period, 30,811 transfusions were administered to 25,448 patients. There were 103 ATRs detected in 81 patients (0.33%; 3.34 reactions per 1000 transfusions, mean age 8.3 ± 5.98 years, 36 females and 45 males). All the reactions were observed within an average of 4 h after the transfusion began. The most common ATRs were allergic reactions (79; 76.6%) and febrile nonhemolytic transfusion reactions (12; 11.6%). All the allergic transfusion reactions occurred within the first hour after the start of the transfusion. Granulocyte concentrates were the blood component associated with the highest ATR rate (2.1%). Conclusion: Within our hospital, pediatric hematology-oncology wards and the stem cell transplantation unit had the most frequent ATR reports; therefore, when transfusions are carried out, increased attention should be given to these units. Educating health staff about the adverse effects of transfusion therapy should increase the awareness and reporting of ATRs in children. [ABSTRACT FROM AUTHOR]

Hemocytes are specialized hemolymph cells found in insects, responsible for a variety of important physiological functions such as immunity, wound healing, and the regulation of the insect's circulatory system. The study of insect hemocytes has become increasingly important in recent years due to the emergence of insect-borne diseases and the need to develop new strategies for insect control. Understanding the biology of insect hemocytes has the potential to lead to the development of novel insecticides and strategies for controlling insect-borne diseases. This study aimed to examine the hemocytes in Bombyx mori to determine if there were any variations in the types of hemocytes to other arthropods. By comprehensively characterizing hemocyte types across developmental stages, this research sheds light on the intricate immune responses and adaptations of the silkworm. Such insights could contribute to the understanding of basic immunological mechanisms in insects. Hemolymph of B. mori was collected, and hemolymph smears were stained with Wright’s stain. In B. mori, four distinct types of hemocyte prohemocyte, plasmatocyte, granulocyte, and oenocytoid were examined. Using light microscopy, the hemocyte types were identified based on their size, the presence or absence of granules, and the ratio of their nucleus to cytoplasm. [ABSTRACT FROM AUTHOR]

Alcoholic liver disease (ALD) significantly affects immune cell function and leads to immunological dysregulation. This study explored the potential of granulocyte colony-stimulating factor (G-CSF) to mitigate the negative effects of alcohol on immune cells in a mouse model of ALD. To investigate the capacity of G-CSF, ALD was induced using a 17-day alcohol-enriched diet, followed by a single G-CSF dose prior to sampling. We focused on the dynamics of peripheral blood mononuclear cells using high-dimensional mass cytometry to detect subtle changes. Alcohol intake reduced the number of B cells, monocytes, dendritic cells, and NK cells while increasing the number of T cells. Notably, G-CSF treatment reversed the alcohol-induced increase in total CD4+ and CD8+ T cell populations. This effect was remarkable in naïve, effector CD4+ T cells and naïve CD8+ T cells. PhenoGraph and FlowSOM analysis further revealed the recovery effect of G-CSF on specific T cell subgroups, including central memory CD8+ T cells and double-negative T cells expressing Ly6chighCD44high, which are adversely affected by alcohol. These results enhance our understanding of the effect of ALD on immune function and suggest that G-CSF is a potential therapeutic agent, laying the foundation for future clinical research.

Typical BCR::ABL1-negative myeloproliferative neoplasms (MPN) are mainly referred to as polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofbrosis (PMF). Granulocytes in MPN patients are involved in their inflammation and form an important part of the pathophysiology of MPN patients. It has been shown that the immunophenotype of granulocytes in MPN patients is altered. We used flow cytometry to explore the immunophenotype of MPN patients and correlate it with clinical parameters. The results showed that PMF patients and PV patients had higher CD15+CD11b+ granulocytes than ET patients and normal controls. When grouped by gene mutation, changes in the granulocyte immunophenotype of MPN patients were independent of the JAK2V617F and CALR mutations. There was no significant heterogeneity in immunophenotype between ET patients and Pre-PMF, and between Overt-PMF and Pre-PMF patients. Granulocytes from some MPN patients showed an abnormal CD13/CD16 phenotype with a significant increase in mature granulocytes on molecular and cytomorphological grounds, and this abnormal pattern occurred significantly more frequently in PMF patients than in ET patients. CD15–CD11b– was negatively correlated with WBC and Hb and positively correlated with DIPSS score, whereas high CD10+ granulocytes were significantly and negatively associated with prognostic system IPSS and DIPSS scores in PMF patients. In conclusion, this study demonstrates the landscape of bone marrow granulocyte immunophenotypes in MPN patients. MPN patients, especially those with PMF, have a significant granulocyte developmental overmaturation phenotype. CD10+ granulocytes may be involved in the prognosis of PMF patients. [ABSTRACT FROM AUTHOR]

Background: The aim of the study was to improve the clinical cognition of leukemia-like reaction caused by voriconazole and granulocyte colony-stimulating factor and to avoid misdiagnosis or delayed diagnosis. Methods: A case of drug analysis of Voriconazole combined with granulocyte colony stimulating factor was retrospectively analyzed and related literature was reviewed. Results: Blood routine of the patient on July 29: WBC 13.48 x 109/L, neutrophil 85.3%, lymphocyte 13.4%, hemoglobin 111 g/L, platelet 285 x 109/L. Vancomycin was given to prevent intracranial infection. Lumbar puncture was performed on July 30, cerebrospinal fluid was sent for routine and biochemical examination, leukocytes were 0.15 x 109/L, monocytes 45%, polynuclear cells 55%, protein 1.172 g/L, Acinetobacter baumannii and Candida clorbicus were detected in sputum culture, vancomycin and meropenem static sites were given to prevent intracranial secondary infection. Fungi were detected in urine culture, and voriconazole was given to prevent fungal infection. Blood routine: White blood cell 0.61 x 109/L, neutrophil 23%, lymphocyte 73.8%, red blood cell 2.65 x 1012/L, hemoglobin 77 g/L, platelet 17 x 109/L, bone marrow was extracted after medication. Bone marrow images show poor myelodysplasia, with granulocytes dominated by proto-early cells. Subsequent flow cytometry, chromosomal karyotype, and fusion gene analysis were performed to exclude the possibility of leukemia. Flow cytometry showed that the proportion of myeloid primordial cells was not high, the granulocytes were mainly at the early and young stage, no abnormal phenotype was observed in erythrocytes, monocytes and NK cells, no obvious mature B lymphocytes were observed, and the ratio of CD4+/CD8+ was decreased. Karyotype results showed that there was no mitotic phase. The results of fusion gene analysis showed that the fusion gene was negative or lower than the detection sensitivity. Voliconazole was stopped first, and granulocyte colony stimulating factor was stopped 3 days later. Two weeks later, blood and bone marrow images basically recovered, white blood cell 7.88 x 109/L, neutrophil 46.3%, lymphocyte 48.2%, hemoglobin 126 g/L, platelet 142 x 109/L, bone marrow hyperplasia active. The proportion of three series is roughly normal. Conclusions: The reason for the occurrence of leukemia-like reaction in this patient was considered to be related to voriconazole and granulocyte colony stimulating factor, cessation of voriconazole and granulocyte colony stimulating factor, and recovery of blood and bone marrow images. In the clinical use of voriconazole and granulocyte colony stimulating factor, close attention should be paid to the drug interaction and individualized medication should be carried out to ensure the safety of medication. [ABSTRACT FROM AUTHOR]

NADPH oxidase is a target of hyperglycemia in type 2 diabetes mellitus (T2DM), which causes dysregulation of enzyme. Alterations in regulation of NADPH oxidase activity mediated receptor and non-receptor signaling in bone marrow granulocytes of mice with obesity-induced T2DM were studied. The animals fed high fat diet (516 kcal/100 g) for 16 weeks. NADPH oxidase-related generation of reactive species (RS) at normo- and hyperthermia was estimated using chemiluminescent analysis. The redox status of the cells was assessed by Redox Sensor Red CC-1. Baseline biochemical indicators in blood (glucose, cholesterol, HDL and LDL levels) were significant higher in T2DM mice versus controls. Using specific inhibitors, signaling mediated by formyl peptide receptors (FPRs) to NADPH oxidase was shown to involve PLC, PKC, cytochrome p450 in both control and T2DM groups and PLA2 in controls. In T2DM regulation of NADPH oxidase activity via mFpr1, a high-affinity receptors, occurred with a significant increase of the role of PKC isoforms and suppression of PLA2 participation. Significant differences between this regulation via mFpr2, low-affinity receptors, were not found. Non-receptor activation of NADPH oxidase with ionomycin (Ca2+ ionophore) or phorbol ester (direct activator of PKC isoforms) did not revealed differences in the kinetic parameters between groups at 37 °C and 40 °C. When these agents were used together (synergistic effect), lower sensitivity of cells to ionophore was observed in T2DM at both temperatures. Redox status in responses to opsonized zymosan was higher in T2DM mice at 37 °C and similar to control levels at 40 °C. ROC-analysis identified Tmax, RS production and effect of opsonized zymosan as the most significant predictors for discriminating between groups. It was concluded that Ca2+-dependent/PKC-mediated regulation of NADPH oxidase activity was altered in BM granulocytes from diabetic mice. [Display omitted] • Regulation of NADPH oxidase activity in diabetic mice granulocytes was studied. • Kinetics of respiratory burst to stimuli and redox status of cells were evaluated. • Ca2+-dependent/PKC-mediated NADPH oxidase activity regulation was altered in T2DM. • Redox status in responses to opsonized zymosan was higher in T2DM mice at 37 °C. • ROC-analysis revealed the most significant predictors to distinguish between groups. [ABSTRACT FROM AUTHOR]

Candida albicans belongs to our commensal mucosal flora and in immune-competent individuals in the absence of epithelial damage, this fungus is well tolerated and controlled by our immune defense. However, C. albicans is an opportunistic microorganism that can cause different forms of infections, ranging from superficial to life-threatening systemic infections. C. albicans is polymorphic and switches between different phenotypes (e.g. from yeast form to hyphal form). C. albicans hyphae are invasive and can grow into tissues to eventually reach circulation. During fungal infections, neutrophils in particular play a critical role for the defense, but how neutrophils are directed toward the invasive forms of fungi is less well understood. We set out to investigate possible neutrophil chemoattractants released by C. albicans into culture supernatants. We found that cell-free culture supernatants from the hyphal form of C. albicans induced both neutrophil chemotaxis and concomitant intracellular calcium transients. Size separation and hydrophobic sorting of supernatants indicated small hydrophilic factors as responsible for the activity. Further analysis showed that the culture supernatants contained high levels of short-chain fatty acids with higher levels from hyphae as compared to yeast. Short-chain fatty acids are known neutrophil chemoattractants acting via the neutrophil free fatty acid receptor 2. In line with this, the calcium signaling in neutrophils induced by hyphae culture supernatants was blocked by a free fatty acid receptor 2 antagonist and potently increased in the presence of a positive allosteric modulator. Our data imply that short-chain fatty acids may act as a recruitment signal whereby neutrophils can detect C. albicans hyphae. [ABSTRACT FROM AUTHOR]

Abstract Background Although some evidence implicates the immune system in migraine attacks, its role during attack-free periods remains largely unexplored. Therefore, we assessed the association between the immune system and migraine status. Methods From the population-based Rotterdam Study, we included 6593 participants who underwent blood sampling and migraine assessments. In the blood samples, we measured white blood-cell-based immune markers. As a marker for the innate immune system, granulocyte and platelet counts were determined, whereas lymphocyte counts were used as a marker for the adaptive immune system. Migraine was assessed using a validated questionnaire based on ICHD-2 criteria. We investigated associations between blood-cell counts and migraine using logistic regression models adjusting for age, sex and other variables. Results Mean age of participants was 65.6 ± 11.2 years and 56.7% were female. The lifetime prevalence of migraine was 15.1% (995/6593). We found no statistically significant associations between granulocyte (odds ratio [OR] per standard deviation increase 1.01 95% Confidence Interval [CI]: 0.93–1.09), platelet (OR 1.01 CI: 0.94–1.09) or lymphocyte counts (OR 1.01 CI: 0.93–1.08) and migraine status. Conclusions Our results do not support an association between white blood-cell-based immunity markers and migraine status.

Introduction: Hematophagous arthropods can acquire and transmit several pathogens of medical importance. In ticks, the innate immune system is crucial in the outcome between vector-pathogen interaction and overall vector competence. However, the specific immune response(s) elicited by the immune cells known as hemocytes remains largely undefined in Ehrlichia chaffeensis and its competent tick vector, Amblyomma americanum. Methods: We utilized injection of clodronate liposome to deplete tick granulocytes combined with infection with E. chaffeensis to demonstrate their essential role in microbial infection. Results: Here, we show that granulocytes, professional phagocytic cells, are integral in eliciting immune responses against commensal and pathogen infection. The chemical depletion of granulocytes led to decreased phagocytic efficiency of tissue-associated hemocytes. We demonstrate that E. chaffeensis can infect circulating hemocytes, and both cell-free plasma and hemocytes from E. chaffeensis-infected ticks can establish Ehrlichia infection in recipient ticks. Lastly, we provide evidence to show that granulocytes play a dual role in E. chaffeensis infection. Depleting granulocytic hemocytes increased Ehrlichia load in the salivary gland and midgut tissues. In contrast, granulocyte depletion led to a reduced systemic load of Ehrlichia. Conclusion: This study has identified multiple roles for granulocytic hemocytes in the control and systemic dissemination of E. chaffeensis infection.

Objectives To investigate laboratory and bone marrow findings that can help predict a diagnosis of hemophagocytic lymphohistiocytosis (HLH) for patients who have demonstrated hemophagocytes (HPCs) in the bone marrow. Methods A total of 57 cases from 48 patients with HPCs present on bone marrow examination were included. The numbers and morphologic characteristics of HPCs with ingested nucleated cells (nHPC) were counted. Pertinent medical history, relevant laboratory values, and flow cytometry data at the time of bone marrow biopsy were collected. Results A total of 24 patients fulfilled diagnostic criteria for HLH, and the remaining 24 patients did not. By using HLH-2004 cutoffs, only hypertriglyceridemia (≥265 mg/dL) was significantly associated with HLH diagnosis. The HLH cases more frequently had nHPC-ingesting granulocytic cells (gHPC) (75.9% vs 24.1%, P  = .009). The percentage of gHPC to all nHPC was also significantly higher in HLH cases (median, 15.4% vs 0%; P  = .0002). Both triglyceride level (area under the curve [AUC]  = 0.88, P  < .0001) and gHPC percentage (AUC = 0.81, P  = .0005) were significant in predicting HLH diagnosis. Finally, no overt immunophenotypic abnormality was noted for 19 HLH cases with available flow cytometry data. Conclusions The presence of hypertriglyceridemia and more frequent gHPC has predictive value for HLH diagnosis in patients with bone marrow HPC. [ABSTRACT FROM AUTHOR]

This 14‐color, 13‐antibody optimized multicolor immunofluorescence panel (OMIP) was designed for deep profiling of neutrophil subsets in various types of human samples to contextualize neutrophil plasticity in a range of healthy and diseased states. Markers present in the OMIP allow the profiling of neutrophil subsets associated with ontogeny, migration, phagocytosis capacity, granule release, and immune modulation. For panel design, we ensured that the commonly available fluorophores FITC/AF488, PE, and APC were assigned to the intracellular subset marker Olfactomedin 4, the maturity and activation marker CD10, and whole blood subset marker CD177, respectively. These markers can be easily replaced without affecting the core identification of neutrophils, enabling antibodies to new neutrophil antigens of interest or for fluorescent substrates to assess different neutrophil functions to be easily explored. Panel optimization was performed on whole blood and purified neutrophils. We demonstrate applications on clinical samples (whole blood and saliva) and experimental endpoints (purified neutrophils stimulated through an in vitro transmigration assay). We hope that providing a uniform platform to analyze neutrophil plasticity in various sample types will facilitate the future understanding of neutrophil subsets in health and disease. [ABSTRACT FROM AUTHOR]

The blood coagulation system is actively involved in the development of cancer. Many solid tumors express tissue factor (TF) that triggers a plasma coagulation cascade leading to an increased risk of cancer-associated thrombosis and venous thrombosis in cancer patients. The main components of the blood clots, platelets are also well known for their role in cancer metastasis as they bind to tumor cells once they enter blood vessels and "shield" them to escape immune surveillance. Platelets also promote adhesion and extravasation of circulating tumor cells into tissues and subsequent metastasis formation. Moreover, being mobile carriers of various growth factors, platelets are actively recruited and, in some cases, engulfed by the tumor, contributing thereby to tumor development and vascularization. Platelet recruitment occurs through both tumor-induced thrombin activation and exposing of platelet-adhesive surfaces by the tumor. Activated in the tumor proximity, platelets attract and induce neutrophil activation and the formation of neutrophil extracellular traps (NETs), thus modulating the tumor microenvironment. Apart from direct interaction, platelets and tumor cells exchange mRNA, miRNA and other regulatory molecules via microvesicles, with platelets serving as "containers" that spread tumor genetic material (circulating nucleic acids) throughout the organism. In this review, we address the molecular mechanisms behind the platelet involvement in the growth and metastasis of solid tumors, and discuss possible options for the pharmacological interruption of platelet–tumor cell interaction. [ABSTRACT FROM AUTHOR]

The perception of circulating granulocytes as cells with a predetermined immune response mainly triggered by pathogens is evolving, recognizing their functional heterogeneity and adaptability, particularly within the neutrophil subset. The involvement of these cells in the pathophysiology of autoimmune uveitis has become increasingly clear, yet their exact role remains elusive. We used an equine model for autoimmune-mediated recurrent pan-uveitis to investigate early responses of granulocytes in different inflammatory environments. For this purpose, we performed differential proteomics on granulocytes from healthy and diseased horses stimulated with IL8, LPS, or PMA. Compared to healthy horses, granulocytes from the recurrent uveitis model significantly changed the cellular abundance of 384 proteins, with a considerable number of specific changes for each stimulant. To gain more insight into the functional impact of these stimulant-specific proteome changes in ERU pathogenesis, we used Ingenuity Pathway Analysis for pathway enrichment. This resulted in specific reaction patterns for each stimulant, with IL8 predominantly promoting Class I MHC-mediated antigen processing and presentation, LPS enhancing processes in phospholipid biosynthesis, and PMA, clearly inducing neutrophil degranulation. These findings shed light on the remarkably differentiated responses of neutrophils, offering valuable insights into their functional heterogeneity in a T-cell-driven disease. Raw data are available via ProteomeXchange with identifier PXD013648. [ABSTRACT FROM AUTHOR]

Background: Although some evidence implicates the immune system in migraine attacks, its role during attack-free periods remains largely unexplored. Therefore, we assessed the association between the immune system and migraine status. Methods: From the population-based Rotterdam Study, we included 6593 participants who underwent blood sampling and migraine assessments. In the blood samples, we measured white blood-cell-based immune markers. As a marker for the innate immune system, granulocyte and platelet counts were determined, whereas lymphocyte counts were used as a marker for the adaptive immune system. Migraine was assessed using a validated questionnaire based on ICHD-2 criteria. We investigated associations between blood-cell counts and migraine using logistic regression models adjusting for age, sex and other variables. Results: Mean age of participants was 65.6 ± 11.2 years and 56.7% were female. The lifetime prevalence of migraine was 15.1% (995/6593). We found no statistically significant associations between granulocyte (odds ratio [OR] per standard deviation increase 1.01 95% Confidence Interval [CI]: 0.93–1.09), platelet (OR 1.01 CI: 0.94–1.09) or lymphocyte counts (OR 1.01 CI: 0.93–1.08) and migraine status. Conclusions: Our results do not support an association between white blood-cell-based immunity markers and migraine status. [ABSTRACT FROM AUTHOR]

Here, retrotransposon-like 1 (RTL1) is introduced as a marker for circulating and tissue neutrophils, tissue macrophages, and tumor-associated macrophages (TAM) and neutrophils (TAN). Anti-RTL1 polyclonal and monoclonal antibodies were produced, and their reactivity was examined by Western blotting (WB), ELISA, and immunostaining of human normal and cancer tissues. The reactivity of the anti-RTL1 antibodies with peripheral blood leukocytes and a panel of hematopoietic cell lines was examined. The generated antibodies specifically detected RTL1 in the WB of the placenta and U937 cells. The polyclonal antibody showed excellent reactivity with tissue-resident macrophages, Hofbauer cells, alveolar and splenic macrophages, Kupffer cells, and inflammatory cells in the tonsil, appendix, and gallbladder. In vitro GM-CSF-differentiated macrophages also showed a high level of intracellular RTL1 expression. TAM and TAN also showed excellent reactivity with this antibody. Almost all circulating granulocytes but not lymphocytes or monocytes expressed RTL1 at their surface. Serial sections of the appendix stained with CD15 and RTL1 and placenta stained with CD68 and RTL1 showed a considerable overlap in RTL1 expression in CD15+ granulocytes and CD68+ macrophages. A small percentage of myelomonocytic cell lines was positive for surface RTL1, while promyelocytic, monocytic, megaloblastic, and lymphoblastic cell lines were negative. Endothelial cells of normal and cancer tissues highly expressed RTL1. RTL1 could be considered a new marker for different normal tissue macrophages, TAM, circulating and tissue neutrophils, and TAN. [ABSTRACT FROM AUTHOR]

To maintain or restore immunological homeostasis, life forms frequently rely on immunomodulators, also known by many terms such as biological response modifiers, immunostimulants, and immune restoratives. Lemon (Citrus limon (L.) Burm) is another well-known source of micronutrients and bioactive phytochemicals that serve as antioxidant against oxidative stress. In this present study, we aimed to examine the immunomodulatory properties of Citrus limon extracts (CLE) on lymphoid and myeloid lineage cells from BALB/c mouse. Six-weeks-old mice were treated by four different doses of CLE, 0, 200, 400, and 800 mg/kg BW for 14 days. Several markers of immune cells were investigated including CD8, CD62L, CD4, B220, VLA-4, TER119, CD55, Gr1, and CD11b antibodies which cover the population of lymphoid and myeloid lineage cells. Flow cytometry analysis was used to identify the specific subset of studied cell population. The results showed that CLE significantly increased the number of Gr1+ granulocyte cells but did not affect other cell types. As a result, we surmised that the components of CLE might have a particular impact on granulocyte cells. Importantly, more investigation is needed to learn how CLE boosts granulocyte cell production. [ABSTRACT FROM AUTHOR]

Background: Crude acetylene fumes generated from Calcium carbide (CaC2) is one of the most widely used source of acetylene gas for occupational gas welding and as an artificial fruit ripener despite numerous health warnings from the government and health groups in Nigeria. Aim: The study was aimed at evaluating the effects of crude acetylene on heavy metals present in serum, haematological indices and histology of the lung tissues. Materials and Methods: Twenty (20) adult male wistar rats were used. The rats were divided into two groups; Group 1 (control) and Group 2 (Test group) (n=10). The animals from the Test group (n = 10) were subsequently exposed to crude acetylene fumes released from the dissolution of 20g of calcium carbide in water for 30 minutes daily for a period of 4 weeks. At the end of the 4th week, the rats were anaesthetized and blood samples were collected for serum heavy metals and haematological analysis and tissues were harvested for histological studies. Results: a significant increase is seen in serum heavy metals (Cr, Fe and Cd) but no significant difference in Pb in the Test group compared with the control group. Also, there was a decrease in percentage granulocytes and histological results show destruction of respiratory epithelium. Also, there were significant damages to the lung tissues which include mild interstitial haemorrhage, bronchiolar ulceration with luminal debris, interstitial oedema, bronchiolar haemorrhage vascular distortion and congestion. Conclusion: Exposure to crude acetylene fumes increases serum heavy metals and can cause disruption of the respiratory epithelium

Background: The therapeutic use of granulocyte transfusions for the treatment of infections in immunocompromised patients has been a controversial practice. Randomized controlled trials suggest that benefit may be provided when a high‐dose product, defined as providing a dose of at least 0.6 × 109/kg, is offered. Here we describe the collection process and granulocyte product yield over a four‐year period at a donation center supplying a large, tertiary academic medical center. Methods: A retrospective chart review was performed for apheresis granulocyte donations collected between 2018 and 2021 following implementation of combined G‐CSF and dexamethasone donor stimulation at our institution. Data collected includes donor demographics, G‐CSF administration timeline, pre‐collection cell counts, product yields, donor adverse events, and post‐transfusion ANC increments. Results: A total of 269 granulocyte units were collected from 184 unique donors. The median neutrophil yield (ANC) following G‐CSF implementation was 7.5 × 1010/unit. The proportion of granulocyte products meeting or exceeding a yield of 4.0 × 1010 per unit was 96.5%. These products resulted in measurable median ANC increment of 550/μL in transfused adult patients (n = 166 transfusions). Discussion: In order to properly assess the effectiveness of granulocyte transfusions in patients, it is necessary to ensure that the products being transfused contain an adequate granulocyte dose. This study demonstrates that the combination of G‐CSF and dexamethasone donor stimulation, followed by apheresis granulocyte collection, is safe and can reliably yield a high‐dose product. Consistent production of high‐dose units allows for better assessment of patient outcomes by reducing dosage variability. [ABSTRACT FROM AUTHOR]

Diacylglycerol kinases (DGKs) play dual roles in cell transformation and immunosurveillance. According to cancer expression databases, acute myeloid leukemia (AML) exhibits significant overexpression of multiple DGK isoforms, including DGKA, DGKD and DGKG, without a precise correlation with specific AML subtypes. In the TGCA database, high DGKA expression negatively correlates with survival, while high DGKG expression is associated with a more favorable prognosis. DGKA and DGKG also feature different patterns of co-expressed genes. Conversely, the BeatAML and TARGET databases show that high DGKH expression is correlated with shorter survival. To assess the suitability of DGKs as therapeutic targets, we treated HL-60 and HEL cells with DGK inhibitors and compared cell growth and survival with those of untransformed lymphocytes. We observed a specific sensitivity to R59022 and R59949, two poorly selective inhibitors, which promoted cytotoxicity and cell accumulation in the S phase in both cell lines. Conversely, the DGKA-specific inhibitors CU-3 and AMB639752 showed poor efficacy. These findings underscore the pivotal and isoform-specific involvement of DGKs in AML, offering a promising pathway for the identification of potential therapeutic targets. Notably, the DGKA and DGKH isoforms emerge as relevant players in AML pathogenesis, albeit DGKA inhibition alone seems insufficient to impair AML cell viability. [ABSTRACT FROM AUTHOR]

The black cutworm, Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae), is widely distributed in the world. It is a polyphagous insect attacking nearly all vegetables and many economic field crops causing a great economic loss. The current research was conducted to evaluate the corruptive effects of entomopathogenic nematodes (EPNs), Steinernema carpocapsae (Weiser) and Heterorhabditis bacteriophora (Poinar) on the hemocyte profile of the A. ipsilon larvae and investigate the interaction between their immune defences and EPN-bacterium complex immunosuppression. For achieving this purpose, the newly moulted 5th instar larvae were infected with LC50 concentrations of S. carpocapsae and H. bacteriophora (21 & 62 IJs/ml, respectively) and the immune reactions were described at certain time intervals post-infection and recorded by photomicrographs. The most important results could be summarized as follows. Five main types of normal circulating hemocytes had been identified, viz., prohemocytes (PRs), plasmatocytes (PLs), granulocytes (GRs), spherulocytes (SPs) and oenocytoids (OEs). The immune responses of A. ipsilon larvae to EPNs were described as encapsulation, nodulation and phagocytosis. On the other hand, the EPN-bacterium complex exhibited different features of suppression on the immune system of A. ipsilon larvae. The major features were considerable deformations of the host hemocytes. At 24 to 48 hrs after infection with S. carpocapsae and H. bacteriophora, toxins produced by their symbiotic bacteria, Xenorhabdus nematophilus and Photorahbdus bacteriophora, respectively, attack A. ipsilon hemocytes leading to death. Because the insects have only innate immune responses against invading pathogenic microbes by immunocytes, the destruction of these hemocytes constitutes a promising approach to the biocontrol of insect pests. Therefore, the tested EPNs S. carpocapsae and H. bacteriophora, can be used in the integrated pest management of the dangerous insect, A. ipsilon. [ABSTRACT FROM AUTHOR]

The role of blood cells as the main components of the physiological defense of insects has been reported. Blood cells are the first factors of the circulatory system that participate in dealing with all kinds of stress and foreign factors. In the present study, four types of blood cells were identified in the hemolymph of the sugar beet weevil Lixus incanescens, which include prohemocytes, granulocytes, plasmatocytes and oenocytoids. Plasmotocytes were the largest hemocytes in terms of size, and the abundance of granulocytes and plasmotocytes in third (70 %) and fourth instar larvae (more 75 %) was higher than other cells. These findings are important as the first step in immunological studies of this insect.

Abstract Background Dysdercus koenigii Fabricius is a polyphagous agricultural pest of paramount importance. Farnesol is a plant-derived open-chain aliphatic sesquiterpenoid alcohol. It is an intermediate in the metabolic pathway of juvenile hormone biosynthesis. The effects of Farnesol on haemocytes of fifth instar nymphs of Dysdercus koenigii were assessed in the present investigation. The early fifth instar nymphs were treated with doses of 0.05 μL, 0.1 μL and 0.2 μL Farnesol topically and day wise change in the total haemocytes was reported. Different types of haemocytes were identified in the haemolymph based on their morphological characteristics. Difference in day wise distribution pattern of haemocytes was evaluated in relation to dose of treatment. Results In normal fifth instar nymphs, age-related variation in the haemocyte count was evident; the haemocyte number increased up to four days and subsequently decreased till six-day when the nymph moulted. The results indicate that Farnesol had an adverse effect on the total number of haemocytes; in treated nymphs the haemocyte count decreased in a dose-dependent manner. The decrease was most prominent in the nymphs treated with a dose of 0.2 μL Farnesol. Based on the morphological characteristics, six different types of haemocytes, viz. prohaemocyte, plasmatocyte, adipohaemocyte, granulocytes, oenocytoids and disintegrated cells were identified in the haemolymph. The percent composition of different haemocytes showed variation depending on the age of the fifth instar nymph and the dose of treatment. In control, the percentages of plasmatocytes decreased up to day 5 and then increased up to 7 days. The percentage of adipohaemocytes increased throughout the nymphal period of the fifth instar, except for a small dip on the fourth day. Granulocytes and oenocytoids showed no clear trend. In Farnesol-treated nymphs, an increase in percent of prohaemocytes was observed in all the treatments. Dose-dependent effects were more prominent after day 5. An increase in plasmatocytes was seen after day 3. A distinct dose-related decrease was reported in adipohaemocytes on all days. Percent of disintegrated cells also showed an increase in all the tested doses of Farnesol on all the days. Granulocytes and oenocytoids showed no definitive trend of change. Conclusions Farnesol is a plant sesquiterpenoid and acts as a precursor in juvenile hormone biosynthesis. The application of Farnesol exogenously may disturb haemocyte patterns which may challenge the immune response of insects. Therefore, Farnesol can potentially be used as an alternative approach in pest management.

Pesticide residues have been reported in hive-stored products for long periods. Larvae of honey bees experience oral or contact exposure to these products during their normal growth and development inside the cells. We analyzed various toxicological, morphogenic, and immunological effects of residue-based concentrations of two fungicides, captan and difenoconazole, on the larvae of worker honey bees, Apis mellifera. Selected concentrations (0.08, 0.4, 2, 10, and 50 ppm) of both fungicides were applied topically at a volume of 1 μL/larva/cell as single and multiple exposures. Our results revealed a continuous, concentration-dependent decrease in brood survival after 24 h of treatment to the capping and emergence stages. Compared to larvae with a single exposure, the multiply exposed youngest larvae were most sensitive to fungicidal toxicity. The larvae that survived higher concentrations, especially multiple exposures, showed several morphological defects at the adult stage. Moreover, difenoconazole-treated larvae showed a significantly decreased number of granulocytes after 1 h of treatment followed by an increase after 24 h of treatment. Thus, fungicidal contamination poses a great risk as the tested concentrations showed adverse effects on the survival, morphology, and immunity of larval honey bees. [ABSTRACT FROM AUTHOR]

Hidradenitis suppurativa (HS) is a chronic inflammatory disease characterized by painful inflamed nodules, abscesses, and pus-draining tunnels appearing in axillary, inguinal, and perianal skin areas. HS lesions contain various types of immigrated immune cells. This study aimed to characterize mediators that support lesional B/plasma cell persistence in HS. Skin samples from several cohorts of HS patients and control cohorts were assessed by mRNA sequencing, quantitative PCR on reverse-transcribed RNA, flow cytometry, and immunohistofluorescence. Blood plasma and cultured skin biopsy samples, keratinocytes, dermal fibroblasts, neutrophilic granulocytes (neutrophils), monocytes, and B cells were analyzed. Complex systems biology approaches were used to evaluate bulk and single-cell RNA sequencing data. Proportions of B/plasma cells, neutrophils, CD8+ T cells, and M 0 and M 1 macrophages were elevated in HS lesions compared to skin of healthy and perilesional intertriginous areas. There was an association between B/plasma cells, neutrophils, and B-cell activating factor (BAFF, aka TNFSF13B). BAFF was abundant in HS lesions, particularly in nodules and abscesses. Among the cell types present in HS lesions, myeloid cells were the main BAFF producers. Mechanistically, granulocyte colony-stimulating factor in the presence of bacterial products was the major stimulus for neutrophils' BAFF secretion. Lesional upregulation of BAFF receptors was attributed to B cells (TNFRSF13C/BAFFR and TNFRSF13B/TACI) and plasma cells (TNFRSF17/BCMA). Characterization of the lesional BAFF pathway revealed molecules involved in migration/adhesion (eg, CXCR4, CD37, CD53, SELL), proliferation/survival (eg, BST2), activation (eg, KLF2, PRKCB), and reactive oxygen species production (eg, NCF1, CYBC1) of B/plasma cells. Neutrophil-derived BAFF supports B/plasma cell persistence and function in HS lesions. [Display omitted] [ABSTRACT FROM AUTHOR]

The perception of circulating granulocytes as cells with a predetermined immune response mainly triggered by pathogens is evolving, recognizing their functional heterogeneity and adaptability, particularly within the neutrophil subset. The involvement of these cells in the pathophysiology of autoimmune uveitis has become increasingly clear, yet their exact role remains elusive. We used an equine model for autoimmune-mediated recurrent pan-uveitis to investigate early responses of granulocytes in different inflammatory environments. For this purpose, we performed differential proteomics on granulocytes from healthy and diseased horses stimulated with IL8, LPS, or PMA. Compared to healthy horses, granulocytes from the recurrent uveitis model significantly changed the cellular abundance of 384 proteins, with a considerable number of specific changes for each stimulant. To gain more insight into the functional impact of these stimulant-specific proteome changes in ERU pathogenesis, we used Ingenuity Pathway Analysis for pathway enrichment. This resulted in specific reaction patterns for each stimulant, with IL8 predominantly promoting Class I MHC-mediated antigen processing and presentation, LPS enhancing processes in phospholipid biosynthesis, and PMA, clearly inducing neutrophil degranulation. These findings shed light on the remarkably differentiated responses of neutrophils, offering valuable insights into their functional heterogeneity in a T-cell-driven disease. Raw data are available via ProteomeXchange with identifier PXD013648.

OBJECTIVE:(a) To evaluate the epidemiology of equine eosinophilic keratoconjunctivitis (EK) in the western United States, (b) to ascertain the efficacy of keratectomy and diamond burr debridement vs medical management alone, (c) to determine the efficacy of various medical therapies, and (d) to further characterize the histopathologic findings of the disease in horses. ANIMALS STUDIED:Twenty-nine horses (47 eyes) diagnosed with EK from 1993 to 2017. PROCEDURE:Retrospective medical record review; owner questionnaire. RESULTS:Average age of presentation was 11 ± 4 years. Warmbloods were significantly overrepresented (P = 0.024). Twenty horses were treated with medical therapy alone, five were treated with superficial lamellar keratectomy, and four were treated with diamond burr debridement. Follow-up data were available for 38 eyes of 23 horses. Median time to resolution for horses treated with either superficial keratectomy or diamond burr debridement (62 days) was not statistically significantly different from those that underwent medical therapy alone (46 days; P = 0.33). Eyes treated with topical steroids had a statistically significant longer median time to resolution (61 days) compared to those that did not receive topical steroid (44 days; P = 0.023). Common histopathologic findings in keratectomy samples included the presence of eosinophils, vascularization, and an eosinophilic membrane spanning areas of ulceration. CONCLUSION:In this population, time to EK resolution was similar for horses treated with medical and surgical management. The use of topical steroids was associated with a prolonged time to resolution. Keratectomy samples from horses with EK had similar findings to those reported in other species.

Abstract Background Immune cell dysfunction plays a central role in sepsis-associated immune paralysis. The transfusion of healthy donor immune cells, i.e., granulocyte concentrates (GC) potentially induces tissue damage via local effects of neutrophils. Initial clinical trials using standard donor GC in a strictly extracorporeal bioreactor system for treatment of septic shock patients already provided evidence for beneficial effects with fewer side effects, by separating patient and donor immune cells using plasma filters. In this ex vivo study, we demonstrate the functional characteristics of a simplified extracorporeal therapy system using purified granulocyte preparations. Methods Purified GC were used in an immune cell perfusion model prefilled with human donor plasma simulating a 6-h treatment. The extracorporeal circuit consisted of a blood circuit and a plasma circuit with 3 plasma filters (PF). PF1 is separating the plasma from the patient’s blood. Plasma is then perfused through PF2 containing donor immune cells and used in a dead-end mode. The filtrated plasma is finally retransfused to the blood circuit. PF3 is included in the plasma backflow as a redundant safety measure. The donor immune cells are retained in the extracorporeal system and discarded after treatment. Phagocytosis activity, oxidative burst and cell viability as well as cytokine release and metabolic parameters of purified GCs were assessed. Results Cells were viable throughout the study period and exhibited well-preserved functionality and efficient metabolic activity. Course of lactate dehydrogenase and free hemoglobin concentration yielded no indication of cell impairment. The capability of the cells to secret various cytokines was preserved. Of particular interest is equivalence in performance of the cells on day 1 and day 3, demonstrating the sustained shelf life and performance of the immune cells in the purified GCs. Conclusion Results demonstrate the suitability of a simplified extracorporeal system. Furthermore, granulocytes remain viable and highly active during a 6-h treatment even after storage for 3 days supporting the treatment of septic patients with this system in advanced clinical trials.

Pesticide residues have been reported in hive-stored products for long periods. Larvae of honey bees experience oral or contact exposure to these products during their normal growth and development inside the cells. We analyzed various toxicological, morphogenic, and immunological effects of residue-based concentrations of two fungicides, captan and difenoconazole, on the larvae of worker honey bees, Apis mellifera. Selected concentrations (0.08, 0.4, 2, 10, and 50 ppm) of both fungicides were applied topically at a volume of 1 µL/larva/cell as single and multiple exposures. Our results revealed a continuous, concentration-dependent decrease in brood survival after 24 h of treatment to the capping and emergence stages. Compared to larvae with a single exposure, the multiply exposed youngest larvae were most sensitive to fungicidal toxicity. The larvae that survived higher concentrations, especially multiple exposures, showed several morphological defects at the adult stage. Moreover, difenoconazole-treated larvae showed a significantly decreased number of granulocytes after 1 h of treatment followed by an increase after 24 h of treatment. Thus, fungicidal contamination poses a great risk as the tested concentrations showed adverse effects on the survival, morphology, and immunity of larval honey bees.

Background: Early Life Stress (ELS) may exert long-lasting biological effects, e.g., on PBMC energy metabolism and mitochondrial respiration. Data on its effect on brain tissue mitochondrial respiration is scarce, and it is unclear whether blood cell mitochondrial activity mirrors that of brain tissue. This study investigated blood immune cell and brain tissue mitochondrial respiratory activity in a porcine ELS model.Methods: This prospective randomized, controlled, animal investigation comprised 12 German Large White swine of either sex, which were weaned at PND (postnatal day) 28–35 (control) or PND21 (ELS). At 20–24 weeks, animals were anesthetized, mechanically ventilated and surgically instrumented. We determined serum hormone, cytokine, and “brain injury marker” levels, superoxide anion (O2•¯) formation and mitochondrial respiration in isolated immune cells and immediate post mortem frontal cortex brain tissue.Results: ELS animals presented with higher glucose levels, lower mean arterial pressure. Most determined serum factors did not differ. In male controls, TNFα and IL-10 levels were both higher than in female controls as well as, no matter the gender in ELS animals. MAP-2, GFAP, and NSE were also higher in male controls than in the other three groups. Neither PBMC routine respiration and brain tissue oxidative phosphorylation nor maximal electron transfer capacity in the uncoupled state (ETC) showed any difference between ELS and controls. There was no significant relation between brain tissue and PBMC, ETC, or brain tissue, ETC, and PBMC bioenergetic health index. Whole blood O2•¯ concentrations and PBMC O2•¯ production were comparable between groups. However, granulocyte O2•¯ production after stimulation with E. coli was lower in the ELS group, and this effect was sex-specific: increased O2•¯ production increased upon stimulation in all control animals, which was abolished in the female ELS swine.Conclusion: This study provides evidence that ELS i) may, gender-specifically, affect the immune response to general anesthesia as well as O2•¯ radical production at sexual maturity, ii) has limited effects on brain and peripheral blood immune cell mitochondrial respiratory activity, and iii) mitochondrial respiratory activity of peripheral blood immune cells and brain tissue do not correlate.

Hematologic malignancies, including multiple myeloma (MM), promote systemic immune dysregulation resulting in an alteration and increased plasticity of myeloid cell subsets. To determine the heterogeneity of the myeloid cell compartment in the peripheral blood of patients with MM, we performed a detailed investigation of the phenotype and function of myeloid subpopulations. We report that a subset of MM patients exhibits a specific myeloid cell phenotype indicative of altered myelopoiesis characterized by significant changes in the properties of circulating granulocytic, monocytic, and eosinophilic populations. The subset, referred to as MM2, is defined by a markedly elevated level of CD64 (FcγRI) on the surface of circulating neutrophils. Compared to healthy controls or MM1 patients displaying intermediate levels of CD64, neutrophils from MM2 patients exhibit a less differentiated phenotype, low levels of CD10 and CXC chemokine receptor 2 (CXCR2), increased capacity for the production of mitochondrial reactive oxygen species, and an expansion of CD16neg immature neutrophil subset. Classical and patrolling monocytes from MM2 patients express elevated levels of CD64 and activation markers. MM2 eosinophils display lower levels of C-C Chemokine receptor 3 (CCR3), Toll-like receptor 4 (TLR4, CD284), and tissue factor (TF, CD142). The MM2 (CD64high) phenotype is independent of age, race, sex, and treatment type. Characteristic features of the MM2 (CD64high) phenotype are associated with myeloma-defining events including elevated involved/uninvolved immunoglobulin free light chain (FLC) ratio at diagnosis. Detailed characterization of the altered myeloid phenotype in multiple myeloma will likely facilitate the identification of patients with an increased risk of disease progression and open new avenues for the rational design of novel therapeutic approaches. [ABSTRACT FROM AUTHOR]

Nikolaos Pournaras,1,2 Anders Andersson,3,4 Melissa A Kovach,1 Médea Padra,5 Karlhans F Che,1,2 Bettina Brundin,1 Shigemi Yoshihara,6 Steven Bozinovski,7 Sara K Lindén,5 Per-Anders Jansson,8 Magnus C Sköld,2,9 Ingemar Qvarfordt,3,8 Anders Lindén1,2 1Division for Lung and Airway Research, Institute of Environmental Medicine, Karolinska Institutet, Stockholm, Sweden; 2Karolinska Severe COPD Center, Department of Respiratory Medicine and Allergy, Karolinska University Hospital, Stockholm, Sweden; 3COPD Center, Department of Respiratory Medicine and Allergology, Sahlgrenska University Hospital, Gothenburg, Sweden; 4COPD Center, Department of Internal Medicine and Clinical Nutrition, Institute of Medicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden; 5Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden; 6Pediatric Allergology and Respiratory Medicine, Department of Pediatrics, Dokkyo Medical University, Mibu, Tochigi, Japan; 7School of Health and Biomedical Sciences, RMIT University, Bundoora, Victoria, Australia; 8Institute of Medicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden; 9Division for Respiratory Medicine, Department of Medicine Solna, Karolinska Institutet, Stockholm, SwedenCorrespondence: Nikolaos Pournaras, Division for Lung and Airway Research, Institute of Environmental Medicine, Karolinska Institutet, Stockholm, Sweden and Karolinska Severe COPD Center, Department of Respiratory Medicine and Allergy, Karolinska University Hospital, Stockholm, Sweden, Email nikolaos.pournaras@ki.sePurpose: Type 2 diabetes mellitus (T2DM) and metabolic syndrome (MetS) are common comorbidities in chronic obstructive pulmonary disease (COPD), but the underlying pathogenic mechanisms are poorly understood. Given that these morbidities all display increased neutrophil mobilization, the current study aimed to address whether glucose homeostasis relates to signs of neutrophil mobilization in COPD.Methods: The study population included healthy non-smokers (HNS) and long-term smokers without (LTS) and with COPD (LTS+COPD). No subject had T2DM or MetS. Serum cotinine was quantified to evaluate current smoking. Capillary blood glucose was measured after overnight fasting and during an oral glucose tolerance test (OGTT). Neutrophils were quantified in blood and bronchoalveolar lavage samples (BAL). The neutrophil-related cytokines IL-36α, -β and -γ were quantified (ELISA) along with IL-6, IL-8, INF-γ and CXCL10 (U-Plex®) in plasma and cell-free BAL fluid (BALF). In addition, we quantified neutrophil elastase (ELISA) and net proteinase activity (substrate assay) in BALF.Results: The LTS+COPD group had lower fasting glucose, greater change in glucose during OGTT and higher neutrophil concentrations in BAL and blood compared with HNS. Fasting glucose correlated in a positive manner with blood neutrophil concentration, forced expiratory volume in 1 second/forced vital capacity ratio (FEV1/FVC) and FEV1 (% of predicted) in LTS+COPD. In this group, the concentration of IL-36α in BALF correlated in a negative manner with fasting glucose, blood neutrophil concentration and FEV1, while the CXCL10 concentration in BALF correlated in a negative manner with glucose at the end of OGTT (120 min). We observed no corresponding correlations for neutrophil elastase, net proteinase or gelatinase activity.Conclusion: In smokers with COPD, altered glucose homeostasis is associated with local and systemic signs of increased neutrophil mobilization, but not with local proteinases. This suggests that other specific aspects of neutrophil mobilization constitute pathogenic factors that affect glucose homeostasis in COPD.Keywords: granulocyte, elastase, IL-36, IP-10, BAL

The present study was conducted to compare the total white blood cells count, granulocytes and phagocytic activity in female rats immunized with lipopolysaccharide (LPS) extracted from two different isolates of Escherichia coli, one normal isolate from the gut of healthy adult and the other was pathogenic strain isolated from patients suffering from urinary tract infection. The study was included 35 female albino rats divided randomly into seven groups, three groups injected with different concentrations of LPS of normal flora of E.coli 90, 120 and 150 microgram/kilogram body weight respectively and the other three groups were injected with the same concentrations of LPS extracted from pathogenic strain of E.coli, the last group (7th) represented the control group which given normal saline only. The results showed that the LPS from normal isolate causes a significant increase in the total number of leukocytes, granulocytes and phagocytic activity in animals immunized with the concentration of 120 µg/kg compared with the control group, and the concentration of 150 µg/kg of the same extract resulted in a high significant increase in phagocytic activity and the concentration of 90 µg/kg did not stimulate any significant differences in all the variables, while immunization with the extract of pathogenic strain increases the total leukocytes at a concentration of 90 µg/kg only with a significant increases in phagocytic activity at all concentration, and it also did not cause any significant differences in the granulocytes count.

Alcoholic liver disease (ALD) significantly affects immune cell function and leads to immunological dysregulation. This study explored the potential of granulocyte colony-stimulating factor (G-CSF) to mitigate the negative effects of alcohol on immune cells in a mouse model of ALD. To investigate the capacity of G-CSF, ALD was induced using a 17-day alcohol-enriched diet, followed by a single G-CSF dose prior to sampling. We focused on the dynamics of peripheral blood mononuclear cells using high-dimensional mass cytometry to detect subtle changes. Alcohol intake reduced the number of B cells, monocytes, dendritic cells, and NK cells while increasing the number of T cells. Notably, G-CSF treatment reversed the alcohol-induced increase in total CD4+ and CD8+ T cell populations. This effect was remarkable in naïve, effector CD4+ T cells and naïve CD8+ T cells. PhenoGraph and FlowSOM analysis further revealed the recovery effect of G-CSF on specific T cell subgroups, including central memory CD8+ T cells and double-negative T cells expressing Ly6chighCD44high, which are adversely affected by alcohol. These results enhance our understanding of the effect of ALD on immune function and suggest that G-CSF is a potential therapeutic agent, laying the foundation for future clinical research. [Display omitted] • We conducted a single-cell-based mass cytometry study to investigate immune cell changes in ALD and G-CSF treated mice. • ALD mice had fewer B cells, monocytes, dendritic cells, and NK cells, but more CD4+ and CD8+ T cells than controls. • G-CSF treatment restored ALD-induced population increases in CD4+ and CD8+ T cells. • This study suggests G-CSF as a potential therapeutic agent for ALD by regulating immune cells and improving liver function. [ABSTRACT FROM AUTHOR]

Hematologic malignancies, including multiple myeloma (MM), promote systemic immune dysregulation resulting in an alteration and increased plasticity of myeloid cell subsets. To determine the heterogeneity of the myeloid cell compartment in the peripheral blood of patients with MM, we performed a detailed investigation of the phenotype and function of myeloid subpopulations. We report that a subset of MM patients exhibits a specific myeloid cell phenotype indicative of altered myelopoiesis characterized by significant changes in the properties of circulating granulocytic, monocytic, and eosinophilic populations. The subset, referred to as MM2, is defined by a markedly elevated level of CD64 (FcγRI) on the surface of circulating neutrophils. Compared to healthy controls or MM1 patients displaying intermediate levels of CD64, neutrophils from MM2 patients exhibit a less differentiated phenotype, low levels of CD10 and CXC chemokine receptor 2 (CXCR2), increased capacity for the production of mitochondrial reactive oxygen species, and an expansion of CD16neg immature neutrophil subset. Classical and patrolling monocytes from MM2 patients express elevated levels of CD64 and activation markers. MM2 eosinophils display lower levels of C-C Chemokine receptor 3 (CCR3), Toll-like receptor 4 (TLR4, CD284), and tissue factor (TF, CD142). The MM2 (CD64high) phenotype is independent of age, race, sex, and treatment type. Characteristic features of the MM2 (CD64high) phenotype are associated with myeloma-defining events including elevated involved/uninvolved immunoglobulin free light chain (FLC) ratio at diagnosis. Detailed characterization of the altered myeloid phenotype in multiple myeloma will likely facilitate the identification of patients with an increased risk of disease progression and open new avenues for the rational design of novel therapeutic approaches.

Filariae are parasitic roundworms, which can cause debilitating diseases such as lymphatic filariasis and onchocerciasis. Lymphatic filariasis, also known as elephantiasis, and onchocerciasis, commonly referred to as river blindness, can lead to stigmatizing pathologies and present a socio-economic burden for affected people and their endemic countries. Filariae typically induce a type 2 immune response, which is characterized by cytokines, i.e., IL-4, IL-5 and IL-13 as well as type 2 immune cells including alternatively activated macrophages, innate lymphoid cells and Th2 cells. However, the hallmark characteristic of filarial infections is a profound eosinophilia. Eosinophils are innate immune cells and pivotal in controlling helminth infections in general and filarial infections in particular. By modulating the function of other leukocytes, eosinophils support and drive type 2 immune responses. Moreover, as primary effector cells, eosinophils can directly attack filariae through the release of granules containing toxic cationic proteins with or without extracellular DNA traps. At the same time, eosinophils can be a driving force for filarial pathology as observed during tropical pulmonary eosinophilia in lymphatic filariasis, in dermatitis in onchocerciasis patients as well as adverse events after treatment of onchocerciasis patients with diethylcarbamazine. This review summarizes the latest findings of the importance of eosinophil effector functions including the role of eosinophil-derived proteins in controlling filarial infections and their impact on filarial pathology analyzing both human and experimental animal studies. [ABSTRACT FROM AUTHOR]

Myeloid leukemia is one of the major causes of deaths among elderly with very poor prognosis. Due to the adverse effects of existing chemotherapeutic agents, plant-derived components are being screened for their anti-leukemic potential. Momordica charantia (bitter gourd) possesses a variety of therapeutic activities. We have earlier demonstrated anti-leukemic activity of acetone extract of M. charantia seeds. The present study reports purification of differentiation inducing principle(s) from further fractionated seed extract (hexane fraction of the acetone extract, Mc2-Ac-hex) using HL-60 cells. Out of the 5 peak fractions (P1–P5) obtained from normal phase HPLC of the Mc2-Ac-hex, only peak fraction 3 (P3) induced differentiation of HL-60 cells as evident from an increase in NBT-positive cells and increased expression of cell surface marker CD11b. The P3 differentiated the HL-60 cells to granulocytic lineage, established by increased CD15 (granulocytic cell surface marker) expression in the treated cells. Further, possible molecular mechanism and the signalling pathway involved in the differentiation of HL-60 cells were also investigated. Use of specific signalling pathway inhibitors in the differentiation study, and proteome array analysis of the treated cells collectively revealed the involvement the of ERK/MAPK mediated pathway. Partial characterization of the P3 by GC–MS analysis revealed the presence of dibutyl phthalate, and derivatives of 2,5-dihydrofuran to be the highest among the 5 identified compounds. This study thus demonstrated that purified differentiation-inducing principle(s) from M. charantia seed extract induce HL-60 cells to granulocytic lineage through ERK/MAPK signalling pathway. [ABSTRACT FROM AUTHOR]

Neutrophils are implicated in the pathogenesis of many diseases involving inflammation. Neutrophils are also critical to host defence and have a key role in the innate immune response to infection. Despite their efficiencies against a wide range of pathogens however, their ability to contain and combat Mycobacterium tuberculosis (Mtb) in the lung remains uncertain and contentious. The host response to Mtb infection is very complex, involving the secretion of various cytokines and chemokines from a wide variety of immune cells, including neutrophils, macrophages, monocytes, T cells, B cells, NK cells and dendritic cells. Considering the contributing role neutrophils play in the advancement of many diseases, understanding how an inflammatory microenvironment affects neutrophils, and how neutrophils interact with other immune cells, particularly in the context of the infected lung, may aid the design of immunomodulatory therapies. In the current review, we provide a brief overview of the mechanisms that underpin pathogen clearance by neutrophils and discuss their role in the context of Mtb and non-Mtb infection. Next, we examine the current evidence demonstrating how neutrophils interact with a range of human and non-human immune cells and how these interactions can differentially prime, activate and alter a repertoire of neutrophil effector functions. Furthermore, we discuss the metabolic pathways employed by neutrophils in modulating their response to activation, pathogen stimulation and infection. To conclude, we highlight knowledge gaps in the field and discuss plausible novel drug treatments that target host neutrophil metabolism and function which could hold therapeutic potential for people suffering from respiratory infections. [ABSTRACT FROM AUTHOR]

Background: Dysdercus koenigii Fabricius is a polyphagous agricultural pest of paramount importance. Farnesol is a plant-derived open-chain aliphatic sesquiterpenoid alcohol. It is an intermediate in the metabolic pathway of juvenile hormone biosynthesis. The effects of Farnesol on haemocytes of fifth instar nymphs of Dysdercus koenigii were assessed in the present investigation. The early fifth instar nymphs were treated with doses of 0.05 μL, 0.1 μL and 0.2 μL Farnesol topically and day wise change in the total haemocytes was reported. Different types of haemocytes were identified in the haemolymph based on their morphological characteristics. Difference in day wise distribution pattern of haemocytes was evaluated in relation to dose of treatment. Results: In normal fifth instar nymphs, age-related variation in the haemocyte count was evident; the haemocyte number increased up to four days and subsequently decreased till six-day when the nymph moulted. The results indicate that Farnesol had an adverse effect on the total number of haemocytes; in treated nymphs the haemocyte count decreased in a dose-dependent manner. The decrease was most prominent in the nymphs treated with a dose of 0.2 μL Farnesol. Based on the morphological characteristics, six different types of haemocytes, viz. prohaemocyte, plasmatocyte, adipohaemocyte, granulocytes, oenocytoids and disintegrated cells were identified in the haemolymph. The percent composition of different haemocytes showed variation depending on the age of the fifth instar nymph and the dose of treatment. In control, the percentages of plasmatocytes decreased up to day 5 and then increased up to 7 days. The percentage of adipohaemocytes increased throughout the nymphal period of the fifth instar, except for a small dip on the fourth day. Granulocytes and oenocytoids showed no clear trend. In Farnesol-treated nymphs, an increase in percent of prohaemocytes was observed in all the treatments. Dose-dependent effects were more prominent after day 5. An increase in plasmatocytes was seen after day 3. A distinct dose-related decrease was reported in adipohaemocytes on all days. Percent of disintegrated cells also showed an increase in all the tested doses of Farnesol on all the days. Granulocytes and oenocytoids showed no definitive trend of change. Conclusions: Farnesol is a plant sesquiterpenoid and acts as a precursor in juvenile hormone biosynthesis. The application of Farnesol exogenously may disturb haemocyte patterns which may challenge the immune response of insects. Therefore, Farnesol can potentially be used as an alternative approach in pest management. [ABSTRACT FROM AUTHOR]

Background/Aim: Although taurolidine is known to exert a wide spectrum of biological actions, its effects on immune cells have not been characterized in detail. In this study, we investigated the ex vivo effects of taurolidine on relevant innate and adaptive immune cell functions. Materials and Methods: Leukocyte functions in whole blood were assessed following treatment with various taurolidine concentrations. Viability of peripheral blood mononuclear cells (PBMCs) and granulocytes was measured using the WST-1 assay. PBMC function was assessed by measuring TNFα and IFNγ production after stimulation with lipopolysaccharide (LPS) or Candida, respectively. Reactive oxygen species (ROS) production by granulocytes was measured in whole blood using luminolenhanced chemiluminescence. Granulocyte degranulation and activation were evaluated by membrane expression of degranulation (CD63, CD66B) and adhesion markers (CD62L, CD11b) using immunofluorescent staining followed by flowcytometric analysis. Results: Taurolidine decreased viability of PBMCs and granulocytes: after 2 h, IC50 concentrations were 500 and 520 µg/ml, respectively. Following prolonged exposure (=24 h) of PBMCs, the IC50 concentrations declined to 40 µg/ml. PBMC cytokine production significantly decreased at taurolidine concentrations below the cytotoxic threshold, whereas no changes in ROS production were observed. The expression of all granulocyte adhesion and degranulation markers increased at concentrations higher than 500 µg/ml (the cytotoxic level of taurolidine). Conclusion: Taurolidine exhibits a dose- and timedependent cytotoxicity toward PBMCs and granulocytes. The effects on PBMCs, as exemplified by a decrease in cytokine production, occurred below the toxic threshold, whereas granulocyte function (ROS production) remained unchanged at these taurolidine concentrations. Granulocyte activation and degranulation markers only increased at cytotoxic taurolidine concentrations. [ABSTRACT FROM AUTHOR]

Filariae are parasitic roundworms, which can cause debilitating diseases such as lymphatic filariasis and onchocerciasis. Lymphatic filariasis, also known as elephantiasis, and onchocerciasis, commonly referred to as river blindness, can lead to stigmatizing pathologies and present a socio-economic burden for affected people and their endemic countries. Filariae typically induce a type 2 immune response, which is characterized by cytokines, i.e., IL-4, IL-5 and IL-13 as well as type 2 immune cells including alternatively activated macrophages, innate lymphoid cells and Th2 cells. However, the hallmark characteristic of filarial infections is a profound eosinophilia. Eosinophils are innate immune cells and pivotal in controlling helminth infections in general and filarial infections in particular. By modulating the function of other leukocytes, eosinophils support and drive type 2 immune responses. Moreover, as primary effector cells, eosinophils can directly attack filariae through the release of granules containing toxic cationic proteins with or without extracellular DNA traps. At the same time, eosinophils can be a driving force for filarial pathology as observed during tropical pulmonary eosinophilia in lymphatic filariasis, in dermatitis in onchocerciasis patients as well as adverse events after treatment of onchocerciasis patients with diethylcarbamazine. This review summarizes the latest findings of the importance of eosinophil effector functions including the role of eosinophil-derived proteins in controlling filarial infections and their impact on filarial pathology analyzing both human and experimental animal studies.

Diacylglycerol kinases (DGKs) play dual roles in cell transformation and immunosurveillance. According to cancer expression databases, acute myeloid leukemia (AML) exhibits significant overexpression of multiple DGK isoforms, including DGKA, DGKD and DGKG, without a precise correlation with specific AML subtypes. In the TGCA database, high DGKA expression negatively correlates with survival, while high DGKG expression is associated with a more favorable prognosis. DGKA and DGKG also feature different patterns of co-expressed genes. Conversely, the BeatAML and TARGET databases show that high DGKH expression is correlated with shorter survival. To assess the suitability of DGKs as therapeutic targets, we treated HL-60 and HEL cells with DGK inhibitors and compared cell growth and survival with those of untransformed lymphocytes. We observed a specific sensitivity to R59022 and R59949, two poorly selective inhibitors, which promoted cytotoxicity and cell accumulation in the S phase in both cell lines. Conversely, the DGKA-specific inhibitors CU-3 and AMB639752 showed poor efficacy. These findings underscore the pivotal and isoform-specific involvement of DGKs in AML, offering a promising pathway for the identification of potential therapeutic targets. Notably, the DGKA and DGKH isoforms emerge as relevant players in AML pathogenesis, albeit DGKA inhibition alone seems insufficient to impair AML cell viability.

Microbubble-mediated ultrasound (MB-US) can be used to realize sonoporation and, in turn, facilitate the transfection of leukocytes in the immune system. Nevertheless, the bio-effects that can be induced by MB-US exposure on leukocytes have not been adequately studied, particularly for different leukocyte lineage subsets with distinct cytological characteristics. Here, we describe how that same set of MB-US exposure conditions would induce heterogeneous bio-effects on the three main leukocyte subsets: lymphocytes, monocytes and granulocytes. MB-US exposure was delivered by applying 1-MHz pulsed ultrasound (0.50-MPa peak negative pressure, 10% duty cycle, 30-s exposure period) in the presence of microbubbles (1:1 cell-to-bubble ratio); sonoporated and non-viable leukocytes were respectively labeled using calcein and propidium iodide. Flow cytometry was then performed to classify leukocytes into their corresponding subsets and to analyze each subset's post-exposure viability, sonoporation rate, uptake characteristics and morphology. Results revealed that, when subjected to MB-US exposure, granulocytes experienced the highest loss of viability (64.0 ± 11.0%) and the lowest sonoporation rate (6.3 ± 2.5%), despite maintaining their size and granularity. In contrast, lymphocytes exhibited the lowest loss of viability (20.9 ± 7.0%), while monocytes had the highest sonoporation rate (24.1 ± 13.6%). For these two sonoporated leukocyte subsets, their cell size and granularity were found to be reduced. Also, they exhibited graded levels of calcein uptake, whereas sonoporated granulocytes achieved only mild calcein uptake. These heterogeneous bio-effects should be accounted for when using MB-US and sonoporation in immunomodulation applications. [ABSTRACT FROM AUTHOR]

Background: Immune cell dysfunction plays a central role in sepsis-associated immune paralysis. The transfusion of healthy donor immune cells, i.e., granulocyte concentrates (GC) potentially induces tissue damage via local effects of neutrophils. Initial clinical trials using standard donor GC in a strictly extracorporeal bioreactor system for treatment of septic shock patients already provided evidence for beneficial effects with fewer side effects, by separating patient and donor immune cells using plasma filters. In this ex vivo study, we demonstrate the functional characteristics of a simplified extracorporeal therapy system using purified granulocyte preparations. Methods: Purified GC were used in an immune cell perfusion model prefilled with human donor plasma simulating a 6-h treatment. The extracorporeal circuit consisted of a blood circuit and a plasma circuit with 3 plasma filters (PF). PF1 is separating the plasma from the patient's blood. Plasma is then perfused through PF2 containing donor immune cells and used in a dead-end mode. The filtrated plasma is finally retransfused to the blood circuit. PF3 is included in the plasma backflow as a redundant safety measure. The donor immune cells are retained in the extracorporeal system and discarded after treatment. Phagocytosis activity, oxidative burst and cell viability as well as cytokine release and metabolic parameters of purified GCs were assessed. Results: Cells were viable throughout the study period and exhibited well-preserved functionality and efficient metabolic activity. Course of lactate dehydrogenase and free hemoglobin concentration yielded no indication of cell impairment. The capability of the cells to secret various cytokines was preserved. Of particular interest is equivalence in performance of the cells on day 1 and day 3, demonstrating the sustained shelf life and performance of the immune cells in the purified GCs. Conclusion: Results demonstrate the suitability of a simplified extracorporeal system. Furthermore, granulocytes remain viable and highly active during a 6-h treatment even after storage for 3 days supporting the treatment of septic patients with this system in advanced clinical trials. [ABSTRACT FROM AUTHOR]

Background: Altered basophil identification markers have been discovered to associate with allergic asthma (AA) in recent years. However, little is known about the expression of basophil markers in blood granulocytes.Aim: To parallel test blood basophils in peripheral blood mononuclear cell (PBMC) and granulocyte populations of patients with AA and AA combined with allergic rhinitis (ARA)Methods: The expressions of surface molecules were determined via flow cytometry. CD123 expressing cells in blood were isolated using a cell sorting technique, and mouse AA models were employed for in vivo study.Results: The numbers of CD123+HLA-DR− cells in the granulocytes of AA and ARA patients markedly increased. However, only 49.7% of CD123+HLA-DR− cells in granulocytes and 99.0% of CD123+HLA-DR− cells in PBMCs were basophils. Almost all CD123+HLA-DR− cells expressed CD63 regardless in granulocytes or PBMC. The numbers of CD63, Fc epsilon receptor I (FcεRI), and CD203c expressing cells markedly enhanced in CD123+HLA-DR− granulocytes of AA and ARA patients. Mean fluorescence intensity (MFI) of CD63 and CD203c expressions on CD123+HLA-DR− PBMC and granulocytes of AA and ARA patients dramatically elevated. House dust mite extract (HDME) and Artemisia sieversiana wild allergen extract (ASWE) enhanced the numbers of CD63+CD123+HLA-DR− granulocytes and PBMC and the MFI of CD203c expression on CD123+HLA-DR− granulocyte of AA and ARA patients. Histamine, tryptase, and PGD2 enhanced proportions of CD123+ KU812 cells. ASWE- and HDME-induced AA mice showed upregulated CD63 expression on basophils. In conclusion, upregulated expressions of CD123, CD203c, CD63, and FcεRIα in PBMC and granulocytes of patients with AA and ARA suggest that CD123+HLA-DR− cells may contribute to the development of AA and ARA.

Studies determining baseline hematological reference intervals (RI) in elasmobranchs are very limited. In this study, blood samples were collected from 94 clinically healthy Nursehound Shark (Scyliorhinus stellaris) maintained under human care. Median (RI) in major leukocyte types were similar to other Carcharhinid sharks as lymphocytes were the predominant leukocyte with 38.0 (28.2–53.5)%, followed by coarse eosinophilic granulocytes with 20.0 (12.2–31.7)%, fine eosinophilic granulocytes with 6.0 (1.2–12.8) %, monocytes with 2.0 (0.0–6.0)%, and neutrophils with 2.0 (0.0–6.0)%. Nursehound Shark produced granulated thrombocytes, which were classified as granulocytes and represented 28.5 (12.4–39.7)% of all leukocytes. Median (RI) manual red blood cell and white blood cell counts were 177.50 (132.50–210.00) x 109 cells/l and 8.26 (5.24–14.23) x 109 cells/l, respectively. Median (RI) plasma chemistry values showed alkaline phosphatase 7.7 (4.2–13.0) U/l, aspartate aminotransferase 7.6 (3.3–17.1) U/l, blood urea nitrogen 281.6 (261.2–305.0) mmol/l, calcium 3.97 (3.59–4.47) mmol/l, total cholesterol 2.04 (1.02–3.91) mmol/l, chloride 233.0 (215.2–259.0) mmol/l, iron 3.79 (1.74–6.93) μmol/l, glucose 0.87 (0.47–1.44 mmol/l), potassium 3.8 (2.9–4.6) mmol/l, sodium 243.0 (227.7–271.0) mmol/l, phosphorus 1.58 (1.13–2.10) mmol/l, total protein 24.0 (20.0–35.0) g/l, and triglycerides 0.97 (0.49–3.35) mmol/l. Creatine kinase, gamma glutamyl transferase, and lactate dehydrogenase levels were below the instrument reading range.