Neutrophil Marker Research Articles

EPS3.05 A neutrophilic inflammation-derived marker of disease

ePS3.05 A neutrophilic inflammation-derived marker of disease

P0302NEUTROPHIL TO LYMPHOCYTE AND PLATELET TO LYMPHOCYTE RATIO IN PATIENTS UNDERGOING KIDNEY TRANSPLANT

Abstract Background and Aims Inflammation plays a central role before and after a kidney transplant recipients (KTR). Despite survival improvement, several factors are associated with poor outcomes after transplantation. In search of a cost effective inflammatory marker neutrophil to lymphocyte ratio (NLR) and the platelet to lymphocyte ratio (PLR) have shown a prognostic value. Therefore, the objective of this study was to determine the association of NLR and PLR and acute kidney allograft dysfunction as markers of inflammatory state in KTR. Method A single center, retrospective study. Our study group included 41 KTR with acute kidney allograft dysfunction from our center at Centro Medico Nacional de Occidente, Jalisco, Mexico. NLR and PLR were collected and evaluated 1 month prior KT, at the time, 6 months and one year after transplantation. Statistical analysis: Data were expressed as the mean ± SD, median and range or frequency, as appropriate. Intergroup comparisons were performed with a chi-squared test for categorical variables and Student’s t test or the Mann-Whitney test for continuous variables. Putative associations between clinical factors, biological factors and mortality were assessed in univariate and multivariate Cox models. Statistical analyses were performed using SPSS v26.0 (IBM Corporation, NY, USA); p < 0.05 was considered statistically significant. Results Mean age was 29.54 ± 7.32 years, and 56% were women. Of those patients, all received living donor kidney transplantation. All patients in the study groups received standardized immunosuppressive regimen consisting of calcineurin inhibitors, mycophenolate mofetil and steroids. Median serum creatinine levels after KT were between 1.11 ± 0.36 mg/dl. The median NLR and PLR levels were significantly higher in the study group, with a median of 3.34 (1.83 –5.14) and 8.65 (343.47 – 360.59), not statistically associated. The best set of predictors in multiple regression analysis of higher levels of NLR were serum albumin (r = -0.432; p = 0.007) and C reactive protein (r = 0.641; p = 0.002). The best set of predictors in multiple regression analysis of higher levels of PLR were hematocrit (r = -0.313; p = 0.055) and serum glucose (r = 0.360; p = 0.026). Conclusion Our data showed that higher values of NLR and PLR are associated with inflammatory markers, leading to the conclusion that this finding must be confirmed with larger, prospective and controlled follow up.

Evaluation of Urinary Neutrophil Gelatinase Associated Lipocalin and Kidney Injury Molecule-1 as Diagnostic Markers for Early Nephropathy in Patients with Type 2 Diabetes Mellitus.

PurposeThe purpose of this study was evaluating the early diagnostic value of two specific tubular markers neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule-1 (KIM-1) in diabetes nephropathy.Patients and MethodsCross-sectional study was carried in three groups of patients from 10/2017 to 10/2018 in Military Hospital 103. Group I included 30 healthy peoples with estimated glomerular filtration rate (eGFR) >60 mL/min/1.73 m2 and urine albumin creatinine ratio (uACR) <30 mg/g. Group II included 30 type 2 diabetic patients having uACR <30 mg/g, eGFR >60 mL/min/1.73 m2. Group III included 30 type 2 diabetic patients having uACR >30 mg/g, eGFR >60 mL/min/1.73 m2.ResultsUrine KIM-1 and NGAL increased progressively from control group (57.29 ± 25.91 pg/mL; 25.71 ± 13.69 ng/mL) to the group of diabetic patients with uACR <30 mg/g (167.06 ± 44.01 pg/mL; 37.42 ± 10.89 ng/mL) and the group of diabetic patients with uACR ≥30 mg/g) (p < 0.05). There were moderate correlations between KIM-1 (r = 0.48, p < 0.05) and NGAL (r = 0.45, p < 0.05) with uACR. There was a mild correlation between KIM-1 and NGAL (r = 0.29, p < 0.05). KIM-1 and NGAL are the independent tests to detect diabetic nephropathy. The sensivity and specificity of KIM-1 with cut-off value of 174.95 pg/mL were 62.37% and 73.48%, respectively; the sensivity and specificity of NGAL with cut-off value of 35.2 ng/mL were 60.45% and 70.37%, respectively.ConclusionKIM-1 and NGAL in urine are independent markers for early diagnostic diabetic nephropathy.

Impacts of dose and length of exposure to boldenone and stanazolol on enzymatic antioxidant systems, myeloperoxidase and NAGase activities, and glycogen and lactate levels in rat liver

We investigated the adverse effects of the anabolic androgenic steroids (AAS) boldenone (BOL) and stanazolol (ST) on the enzymatic antioxidant systems of the rat liver. Male Wistar rats were divided in three protocols (P): PI, 5 mg/kg BOL or ST once a week for 4 weeks; PII, 2.5 mg/kg BOL or ST once a week for 8 weeks; PIII, 1.25 mg/kg BOL or ST once a week for 12 weeks. AAS were administered intramuscularly (0.2 ml, olive oil vehicle) once a week in all protocols. Activities of the enzymes glutathione peroxidase (GPx), glutathione S-transferase (GST), and glutathione reductase (GR), superoxide dismutase (SOD), catalase (CAT), were investigated. We assessed the content of hydrogen peroxide (H2O2), glycogen and lactate; and enzyme markers of neutrophils (myeloperoxidase, MPO) and macrophages (NAGase). PI and PII altered the SOD and CAT activities and increased the H2O2 content. PI led to increases in the MPO and NAGase activities. In contrast, changes in GPx, GST and, GR were observed under PII and, to a greater extend, under PIII. Following PIII, GPx, GR, and GST exhibited reduced activities. All protocols altered the glycogen and lactate content. The use of high doses of AAS for a short duration first alters SOD/CAT activity. In contrast, at lower doses of AAS for long periods is associated with changes in the glutathione system. Protocols with high doses of AAS for a short duration exert the most deleterious effects on redox status, markers of cellular infiltration, and the metabolic functioning of hepatic tissues.

Neutrophil Phenotypes in Coronary Artery Disease.

Clinical evidence indicates that innate immune cells may contribute to acute coronary syndrome (ACS). Our prospective study aimed at investigating the association of neutrophil phenotypes with ACS. 108 patients were categorized into chronic stable coronary artery disease (n = 37), unstable angina (UA) (n = 19), Non-ST-Elevation Myocardial Infarction (NSTEMI) (n = 25), and ST-Elevation Myocardial Infarction (STEMI) (n = 27). At the time of inclusion, blood neutrophil subpopulations were analysed by flow cytometry. Differential blood cell count and plasma levels of neutrophilic soluble markers were recorded at admission and, for half of patients, at six-month follow-up. STEMI and NSTEMI patients displayed higher neutrophil count and neutrophil-to-lymphocyte ratio than stable and UA patients (p < 0.0001), which normalized at six-month post-MI. Atypical low-density neutrophils were detected in the blood of the four patient groups. STEMI patients were characterized by elevated percentages of band cells compared to the other patients (p = 0.019). Multivariable logistic regression analysis revealed that plasma levels of total myeloperoxidase was associated with STEMI compared to stable (OR: 1.434; 95% CI: 1.119–1.837; P < 0.0001), UA (1.47; 1.146–1.886; p = 0.002), and NSTEMI (1.213; 1.1–1.134; p = 0.0001) patients, while increased neutrophil side scatter (SSC) signal intensity was associated with NSTEMI compared to stable patients (3.828; 1.033–14.184; p = 0.045). Hence, changes in neutrophil phenotype are concomitant to ACS.

Effect of first-line spartalizumab + dabrafenib + trametinib on immunosuppressive features detected in peripheral blood and clinical outcome in patients (pts) with advanced BRAF V600–mutant melanoma.

10034 Background: Spartalizumab + dabrafenib + trametinib has previously shown a high response rate of 78% (28 of 36 pts), with a complete response (CR) rate of 42%. A correlative analysis of gene expression signatures (GES)/pathways using whole-transcriptome RNA-seq data from tissue showed that pts with a CR had significantly lower expression levels of immunosuppressive factors in the tumor microenvironment (TME) at baseline (BL). Here we analyze BL peripheral blood markers in the same cohort of pts to assess whether liquid markers can also predict response and clinical outcome to spartalizumab + dabrafenib + trametinib. Methods: The Phase III COMBI-i study (NCT02967692) is evaluating spartalizumab + dabrafenib + trametinib in pts with previously untreated BRAF V600–mutant unresectable or metastatic melanoma. In parts 1 (safety run-in; n = 9) and 2 (biomarker cohort; n = 27), blood and tissue samples were collected at BL, on treatment after 2-3 wk and 8-12 wk, and at disease progression. Lactate dehydrogenase (LDH) and other blood-based markers (including cytokine profiling [n = 45] and blood RNA-seq [114 signatures]) were assessed in all 36 pts. Pts were divided into 2 groups of 24 and 12 pts based on progression-free survival (PFS) of &gt; 1 or &lt; 1 y. Results: In addition to LDH, previously described blood markers such as neutrophil to lymphocyte ratio (NLR) and plasma IL-8 were identified among other neutrophil and immunosuppressive features as top candidates associated with PFS &gt; 1 y. Low plasma IL-8 levels were also associated with CR, and multivariate models suggested that IL-8 may add independent predictive value to LDH and NLR for PFS &gt; 1 y and CR. Pts with high IL-8 levels in the circulation were characterized by high neutrophil chemokine signaling (ρ = 0.553) and high neutrophil markers (ρ = 0.466) in the tumor as measured by RNA-seq GES levels. We observed a decrease in plasma IL-8 levels from BL upon treatment with spartalizumab + dabrafenib + trametinib. Conclusions: Our peripheral blood marker analysis confirmed recent findings from tissue samples that intratumoral immunosuppressive features may preclude a CR and are associated with poor outcomes. High BL plasma IL-8 levels may be associated with an immunosuppressive TME. Further validation is warranted; the randomized placebo-controlled part 3 of COMBI-i is ongoing. Clinical trial information: NCT02967692.

Amplification of transglutaminase 2 enhances tumor-promoting inflammation in gastric cancers

Tumor-promoting inflammation is a hallmark of cancer and is highly associated with tumor progression, angiogenesis, and metastasis. Tumor-associated macrophages (TAMs) are major drivers of tumor-promoting inflammation, but due to the complexity of the tumor microenvironment, the detailed regulatory mechanisms are still under investigation. Here, we investigated a novel role for transglutaminase 2 (TGM2) in the development of tumor-promoting inflammation and recruitment of TAMs to gastric cancer (GC) tissues. When estimated by array comparative genomic hybridization and droplet digital PCR, the copy numbers of the TGM2 gene were amplified in 13.6% (14/103) of GC patients and positively associated with TGM2 expression. Gene set enrichment analysis of expression microarray data for GC samples with high or low TGM2 expression showed that increased TGM2 expression was associated with tumor-promoting inflammation in GC. In addition, the expression of TGM2 was correlated with the expression of markers for macrophages, neutrophils, blood vessels, and lymphatic vessels. Overexpression of TGM2 in GC cells augmented the IL-1β-induced secretion of macrophage-recruiting chemokines and NF-κB activation. TGM2 protein levels were associated with the expression levels of the macrophage marker CD163 in human GC tissue samples. Moreover, GC patients with high expression of TGM2 had a worse prognosis than those with low expression of TGM2. These results suggest TGM2 as a novel regulator of the tumor microenvironment of GC and provide a promising target for constraining tumor-promoting inflammation.

Low density neutrophils as a potential biomarker of leprosy severity

Abstract Erythema Nodosum Leprosum (ENL) is a recurrent acute inflammatory complication of leprosy affecting up to 50% of patients with multibacillary (MB) leprosy, and is treated with thalidomide. Although ENL is described as an immune reaction mediated by neutrophils, studies showing the direct role of neutrophils in ENL are still neglected. One subpopulation of low-density neutrophils (LDNs), present within the fraction of peripheral blood mononuclear cells (PBMC), is associated in the pathogenesis and severity of diseases like sepsis, lupus and tuberculosis. In this study, we characterized circulating LDNs in leprosy, highlighting their involvement in the development of ENL. First, we observed segmented/hypersegmented cells morphologically compatible to neutrophils in PBMC of nonreactional and ENL patients. By flow cytometry, we confirmed the elevated frequency of circulating LDNs (CD14−CD15+) in ENL patients compared to nonreactional. Also, the pattern of expression of neutrophil activation markers suggests a greater activation of LDNs during ENL. Treatment with thalidomide did not interfered the frequency of LDNs, but reduced their activation. Serum levels of MMP-9 (neutrophilic degranulation marker) were correlated with the frequency of LDNs. Finally, we revealed that Mycobacterium leprae (ML) induce generation of LDNs in whole blood and purified normal density neutrophils of healthy individuals in vitro. In brief, we showed that activated LDNs are elevated in the circulation of ENL patients, and may be originated by degranulation caused by ML stimulus. These results point the neutrophilic involvement in leprosy especially on ENL favoring the systemic inflammation and suggest LDNs as a biomarker in ENL development.

CD44‐Targeted Monoclonal Antibody is Anti‐Inflammatory in an In‐Vivo Model of Monosodium Urate Crystal Induced Peritonitis

CD44‐Targeted Monoclonal Antibody is Anti‐Inflammatory in an <i>In‐Vivo</i> Model of Monosodium Urate Crystal Induced Peritonitis

SITE-1 protease ablation in the osterix-lineage results in bone marrow dysplasia with neutrophilia coupled to osteopenia

SITE-1 protease ablation in the osterix-lineage results in bone marrow dysplasia with neutrophilia coupled to osteopenia

The Influence of Salt Loading on the Kidney Injury Biomarker Neutrophil Gelatinase‐Associated Lipocalin in Healthy Young Adults

PurposeThe kidneys play a critical role in blood pressure (BP) regulation. In hypertensive patients and in rodent models, long‐term high dietary sodium (Na+) causes a reduction in kidney function (reduced glomerular filtration rate; GFR). Rodent studies demonstrate that high dietary Na+ increases levels of novel kidney injury biomarkers, independent of changes in BP and GFR. However, it is unclear if short‐term high Na+ increases kidney injury biomarkers in healthy humans. Therefore, we tested the hypothesis that short‐term high Na+ intake increases urinary excretion of the novel kidney injury marker Neutrophil Gelatinase‐Associated Lipocalin (NGAL) in healthy, young adults.MethodsTwenty participants (12 M/8 F; age: 24±4 years; BMI: 23.1±0.6 kg/m2; BP: 112 ±10/64 ±9 mmHg, mean ± SD) participated in a double‐blind, placebo‐controlled, randomized, crossover study. For 10‐days, participants were asked to consume a 2,300 mg/day Na+ diet and supplement with salt (3,900 mg/day of Na+) or placebo (dextrose) capsules. The conditions were separated by at least two weeks and female participants were tested during the placebo phase of their oral contraceptive cycle. Participants collected their urine for the final 24 hours of each supplementation period to assess urine flow rate, and Na+ excretion to verify diet adherence. Brachial BP was measured after at least five minutes of rest. We measured urine and serum creatinine (kinetic modification via the Jaffe reaction) to estimate GFR via creatinine clearance. We used enzyme‐linked immunosorbent assays (Toronto Bioscience) to measure urinary NGAL. Statistical analysis included paired t‐tests and Wilcoxon matched‐pairs signed rank tests.ResultsCompared to placebo, Na+ supplementation increased urinary Na+ excretion (139.9±68.4 vs. 282.5±69.8 mmol/24 hours, p&lt;0.01) but there was no difference in mean arterial BP (77±7 vs. 77±6 mmHg, p=0.64). Serum creatinine concentrations (0.78±0.12 vs. 0.77±0.09 mg/dL, p=0.55) were not different between conditions. However, compared to placebo, Na+ supplementation increased creatinine clearance (110.5±32.9 vs. 145.0±24.9 mL/min, p&lt;0.01) and increased urinary NGAL excretion indexed to urine flow rate (16.9±21.9 vs. 28.4±36.1 ng/min, p=0.02).DiscussionOur preliminary data suggests that salt loading does not alter serum creatinine concentration but increases creatinine clearance and urinary excretion of the kidney injury marker NGAL in healthy young adults.Support or Funding InformationFundingSupported by NIH grants R01HL128388 and K01HL147998, and ACSM Foundation Doctoral Student Research Grant 17‐00521

Annexin A1 Expression Capacity as a Determinant for Disease Severity in Crohn’s Disease

Introduction: Crohn’s disease (CD) is characterized by relapsing intestinal inflammation. The anti-inflammatory protein annexin A1 (ANXA1) has been linked to inflammatory processes in the gut. Objective: To examine ANXA1 expression patterns in the inflamed intestine of patients with CD and associate ANXA1 expression capacity with disease characteristics. Methods: Surgical specimens of patients with CD operated between 2003 and 2015 were examined. Immunohistochemistry and immunofluorescence were performed to delineate ANXA1 expression. Those with pronounced ANXA1 expression were included in further analysis by qPCR. ANXA1 mRNA expression ratio of the inflamed to non-inflamed tissue was determined and defined as expression capacity of the tissue. Depending on their expression capacity, patients were divided into 2 groups (ANXA1-low vs. ANXA1-high), which were associated with clinical characteristics. Results: Immunohistochemical ANXA1 expression was localized in inflamed regions of the intestine. In immunofluorescence, ANXA1 costained with myeloperoxidase as neutrophil marker, CD4 and CD8 as T cell marker but not CD20 as B cell marker or CD68 as macrophage marker. In qPCR, ANXA1 mRNA expression was upregulated by 20-fold in inflamed to noninflamed tissues. Patients with higher intrinsic ANXA1 expression capacity had significantly less severity of inflammation. Furthermore, the ANXA1-high group had significantly more locally restricted disease (p = 0.0070), more stricturating disease (p = 0.0037), and was less frequently treated by preoperative steroid therapy (p = 0.030). Conclusions: ANXA1 expression was strongly associated with intestinal inflammation and expressed in T cells and neutrophils of the CD tissues. Patients with higher intrinsic ANXA1 expression capacity of the inflamed tissue presented milder inflammatory changes and indolent clinical course.

Neutrophil Lymphocyte Ratio and Platelet Lymphocyte Ratio as Diagnostic and Prognostic Markers for Hepatitis C Virus – Related Hepatocellular Carcinoma in Egyptian Patients

Abstract Background hepatocellular carcinoma (HCC), a highly prevalent and lethal cancer, it is the sixth most common cancer and the third leading cause of cancer-related death worldwide. Aim of the work to evaluate the role of inflammatory markers Neutrophil lymphocyte ratio and Platelet lymphocyte ratio (NLR &amp; PLR) as biomarkers for diagnosis of HCV related HCC. Patients and Methods this study was conducted in tropical medicine department, HCC clinic Ain-shams university hospitals. Our study included 174 candidates who were divided into three groups. Group A included 60 Patients with HCV-related HCC diagnosed according to American Association for the Study of Liver Diseases (AASLD) guidelines; HCC patients were subdivided into three subgroups According to BCLC Results group A included 114 Patients with HCV-related HCC, which was subdivided into three subgroups according to BCLC. Group 1 included 30 patients underwent Radio-frequency ablation (RF), 76.67% of them were males while 23.33% were females with mean age 57.433. Group 2 included 41 patients underwent Trans-arterial chemo-embolization (TACE), 80.49% were males while 19.51% were females with mean age 60.268. Group 3 included 43 Patients with BCLC stage C and D For best supportive care, 76.74% were males while 23.26% were females with mean age 60.372. Male to female ratio in HCC patients was 3.56:1. Conclusion AFP remains the gold standard marker for diagnosis of HCC. NLR and PLR has no role as early prognostic markers for HCC.

Markers of Disease Severity Amongst Homozygous Sickle Cell Anaemia Attending Outpatient At Bowen University Teaching Hospital Ogbomoso, Nigeria.

Introduction: Genetic, cellular and molecular modifiers are responsible for the notoriously variable sickle cell phenotype. Haemoglobin F is a principal modulator of the SCD phenotype. Haemoglobin F inhibits the polymerization of Haemoglobin S and ameliorates the secondary effects of sickling. We measured the Haemoglobin F(HbF) in our population and compared it with other markers associated with clinical severity and clinical status of the patients.&#x0D; Methods: we randomly selected 40 Hemoglobin S(HbS) patients who have never taken hydroxyurea. We measured hemoglobin F levels, packed cell volume (PCV) and reticulocyte count to serve as markers for hemolysis, neutrophils, platelets and MCHC to serve as markers for clinical severity. We searched for a relationship between these laboratory features and frequency of vaso-occlusive crises, transfusion history and number of hospital visits per year. They were compared with twenty healthy Hemoglobin A (HbA) controls.&#x0D; Results: Packed cell volume was significantly lower and the reticulocyte was significantly higher in the hemoglobin S compared with controls. The platelet count of the sickle cell anaemia patients was more than twice the number of the controls. The mean hemoglobin F level was 7.1± 3.5 %. The hemoglobin F was negatively correlated with the platelet count but positively correlated with the total white cell count and haematocrit but there was no significant correlation between hemoglobin F and clinical features&#x0D; Conclusion: The fetal hemoglobin level may not be the only modifier responsible for phenotype in population of hemoglobin S

A Precision Medicine Approach Uncovers a Unique Signature of Neutrophils in Patients With Brushite Kidney Stones

IntroductionWe have previously found that papillary histopathology differs greatly between calcium oxalate and brushite stone formers (SF); the latter have much more papillary mineral deposition, tubular cell injury, and tissue fibrosis.MethodsIn this study, we applied unbiased orthogonal omics approaches on biopsied renal papillae and extracted stones from patients with brushite or calcium oxalate (CaOx) stones. Our goal was to discover stone type-specific molecular signatures to advance our understanding of the underlying pathogenesis.ResultsBrushite SF did not differ from CaOx SF with respect to metabolic risk factors for stones but did exhibit increased tubule plugging in their papillae. Brushite SF had upregulation of inflammatory pathways in papillary tissue and increased neutrophil markers in stone matrix compared with those with CaOx stones. Large-scale 3-dimensional tissue cytometry on renal papillary biopsies showed an increase in the number and density of neutrophils in the papillae of patients with brushite versus CaOx, thereby linking the observed inflammatory signatures to the neutrophils in the tissue. To explain how neutrophil proteins appear in the stone matrix, we measured neutrophil extracellular trap (NET) formation—NETosis—and found it significantly increased in the papillae of patients with brushite stones compared with CaOx stones.ConclusionWe show that increased neutrophil infiltration and NETosis is an unrecognized factor that differentiates brushite and CaOx SF and may explain the markedly increased scarring and inflammation seen in the papillae of patients with brushite stones. Given the increasing prevalence of brushite stones, the role of neutrophil activation in brushite stone formation requires further study.

Are Serum-Based Markers Going to Replace Faecal Neutrophil Markers in Ulcerative Colitis?

Are Serum-Based Markers Going to Replace Faecal Neutrophil Markers in Ulcerative Colitis?

Neutrophil Chemotaxis and NETosis in Murine Chronic Liver Injury via Cannabinoid Receptor 1/ Gαi/o/ ROS/ p38 MAPK Signaling Pathway.

Neutrophils play an essential role in the control of inflammatory diseases. However, whether cannabinoid receptors (CBs) play a role in neutrophil chemotaxis and NETosis in sterile liver inflammation remains unknown. The expression of marker genes on neutrophils was characterized by FACS, immunofluorescence, qRT-PCR, and Western blot. The amount of neutrophils was significantly elevated from 7 days and reached the peak at 2 weeks in carbon tetrachloride (CCl4)-treated mouse liver. The mRNA expression of neutrophil marker Ly6G had positive correlation with CB1 and CB2 expression in injured liver. In vitro CBs were abundantly expressed in isolated neutrophils and CB1 agonist ACEA promoted the chemotaxis and cytoskeletal remodeling, which can be suppressed by CB1 antagonist AM281. Moreover, ACEA induced NETosis, myeloperoxidase release from lysosome and ROS burst, indicating neutrophil activation, via Gαi/o. Conversely, CB2 agonist JWH133 had no effect on neutrophil function. ROS and p38 MAPK signaling pathways were involved in CB1-mediated neutrophil function, and ROS was upstream of p38 MAPK. CB1 blockade in vivo significantly attenuated neutrophil infiltration and liver inflammation in CCl4-treated mice. Taken together, CB1 mediates neutrophil chemotaxis and NETosis via Gαi/o/ROS/p38 MAPK signaling pathway in liver inflammation, which represents an effective therapeutic strategy for liver diseases.

Changes in macrophage inflammatory protein-1 (MIP-1) family members expression induced by traumatic brain injury in mice.

A deep knowledge of the profound immunological response induced by traumatic brain injury (TBI) raises the possibility of novel therapeutic interventions. Existing studies have highlighted the important roles of C-C motif ligands in the development of neuroinflammation after brain injury; however, the participation of macrophage inflammatory protein-1 (MIP-1) family members in this phenomenon is still undefined. Therefore, the goal of our study was to evaluate changes in macrophage inflammatory protein-1 (MIP-1) family members (CCL3, CCL4, and CCL9) and their receptors (CCR1 and CCR5) in a mouse model of TBI (induced by controlled cortical impact (CCI)). We also investigated the pattern of activation of immunological cells (such as neutrophils, microglia and astroglia), which on one hand express CCR1/CCR5, and on the other hand might be a source of the tested chemokines in the injured brain. We investigated changes in mRNA (RT-qPCR) and/or protein (ELISA and Western blot) expression in brain structures (the cortex, hippocampus, thalamus, and striatum) at different time points (24 h, 4 days, 7 days, 2 weeks, and/or 5 weeks) after trauma. Our time-course studies revealed the upregulation of the mRNA expression of all members of the MIP-1 family (CCL3, CCL4, and CCL9) in all tested brain structures, mainly in the early stages after injury. A similar pattern of activation was observed at the protein level in the cortex and thalamus, where the strongest activation was observed 1 day after CCI; however, we did not observe any change in CCL3 in the thalamus. Analyses of CCR1 and CCR5 demonstrated the upregulation of the mRNA expression of both receptors in all tested cerebral structures, mainly in the early phases post injury (24 h, 4 days and 7 days). Protein analysis showed the upregulation of CCR1 and CCR5 in the thalamus 24 h after TBI, but we did not detect any change in the cortex. We also observed the upregulation of neutrophil marker (MPO) at the early time points (24 h and 7 days) in the cortex, while the profound activation of microglia (IBA-1) and astroglia (GFAP) was observed mainly on day 7. Our findings highlight for the first time that CCL3, CCL4, CCL9 and their receptors offer promising targets for influencing secondary neuronal injury and improving TBI therapy. The results suggest that the MIP-1 family is an important target for pharmacological intervention for brain injury.

Human umbilical cord mesenchymal stem cells alleviate inflammatory bowel disease by inhibiting ERK phosphorylation in neutrophils.

Inflammatory bowel disease (IBD) can be caused by a variety of factors, including hereditary and environmental influences, that lead to dysfunction of the intestinal immune system. Mesenchymal stem cells (MSCs) exhibit important regulatory roles in relieving inflammation and repairing damaged tissues. Although neutrophils are important participants in the development of inflammatory reactions, they are also essential for maintaining intestinal balance during the process of mitigation of IBD by MSCs. Here, we constructed a dextran sulfate sodium (DSS)-induced mouse IBD model and evaluated the effects of treatment with human umbilical cord MSCs. Mouse body weight, faecal traits, colon/spleen gross morphology, tissue histology and immunohistochemical staining, and inflammatory factors were analysed. Magnetic beads were used to sort infiltrating neutrophils from intestinal tissues, and their phenotypes were identified. The neutrophil inflammatory environment was also simulated in vitro, and signalling pathways involved in MSC regulation of neutrophil phenotype were analysed. Human umbilical cord MSCs effectively alleviated DSS-induced weight loss, colon shortening, and intestinal mucosal injury, and reduced clinical disease activity index. The number of neutrophils that infiltrated the intestines of mice treated with human umbilical cord MSCs were decreased and polarised toward the N2 phenotype; at the same time, ERK phosphorylation was inhibited. In vitro experiments showed that addition of the ERK phosphorylation inhibitor, PD98059, down-regulated the expression of N1 neutrophils, while up-regulating that of N2 neutrophils. The colon tissues from patients with IBD were infiltrated with neutrophils. Further, relative to healthy controls, the markers of N1 neutrophils (ICAM-1, FAS, and CCL3) were highly expressed in colon tissues from patients with IBD, whereas the markers of N2 neutrophils (VEGF, CCL2, and CXCR4) were almost undetectable. In conclusion, during alleviation of IBD, human umbilical cord MSCs polarise neutrophils toward the "N2" phenotype by inhibiting activation of ERK signalling.

Neutrophil Subsets in Periodontal Health and Disease: A Mini Review.

Neutrophils are amongst the most abundant immune cells within the periodontal tissues and oral cavity. As innate immune cells, they are first line defenders at the tooth-mucosa interface, and can perform an array of different functions. With regard to these, it has been observed over many years that neutrophils are highly heterogeneous in their behavior. Therefore, it has been speculated that neutrophils, similarly to other leukocytes, exist in distinct subsets. Several studies have investigated different markers of neutrophils in oral health and disease in recent years in order to define potential cell subsets and their specific tasks. This research was inspired by recent advancements in other fields of medicine in this field. The aim of this review is to give an overview of the current evidence regarding the existence and presence of neutrophil subsets and their possible functions, specifically in the context of periodontitis, gingivitis, and periodontal health.