Degranulation Products Research Articles

Immune Response Dynamics and Biomarkers in COVID-19 Patients.

The immune response dynamics in COVID-19 patients remain a subject of intense investigation due to their implications for disease severity and treatment outcomes. We examined changes in leukocyte levels, eosinophil activity, and cytokine profiles in patients hospitalized with COVID-19. Serum samples were collected within the first 10 days of hospitalization/confirmed infection and analyzed for eosinophil granule proteins (EGP) and cytokines. Information from medical records including comorbidities, clinical symptoms, medications, and complete blood counts were collected at the time of admission, during hospitalization and at follow up approximately 3 months later. Serum levels of eotaxin, type 1 and type 2 cytokines, and alarmin cytokines were elevated in COVID-19 patients, highlighting the heightened immune response (p < 0.05). However, COVID-19 patients exhibited lower levels of eosinophils and eosinophil degranulation products compared to hospitalized controls (p < 0.05). Leukocyte counts increased consistently from admission to follow-up, indicative of recovery. Attenuated eosinophil activity alongside elevated chemokine and cytokine levels during active infection, highlights the complex interplay of immune mediators in the pathogenesis COVID-19 and underscores the need for further investigation into immune biomarkers and treatment strategies.

Eosinophilic esophagitis: absolute eosinophilic count, peak eosinophilic count, and potential biomarkers of eosinophilic degranulation products-an in-depth systematic review.

Eosinophilic esophagitis is a chronic inflammatory disorder, often relapsing. There is an increasing need to develop new alternative diagnostic and monitoring methods on a critical basis, which will provide samples through none or minimally invasive procedures. This study aims to identify and document the types and roles of potential biomarkers in eosinophilic esophagitis released by eosinophils as well as the potential relationship to the peak eosinophilic count and the degree of degranulation of in situ eosinophils (DGE/DGE + NDGE: degranulated eosinophils/degranulated eosinophils and non-degranulated eosinophils). This is the first in-depth systematic review study using PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) parameters involving a literature search of academic databases (PubMed, Scopus, Medline, Google Scholar, and Cochrane Database, 2011-2022) targeting specifically the eosinophilic counts and ratio, and the eosinophilic degranulation products as potential biomarkers. Data were extracted from ten selected studies and presented on a spreadsheet. The studies show the ability to detect eosinophilic and non-eosinophilic degranulation products, and absolute eosinophilic count in samples, including blood and urine, thereby serving as potential surrogates in making the diagnosis or monitoring disease progression in the future. There is an obvious paucity of studies that correlate potential biomarkers to the degree of degranulation of in situ eosinophils. A few minimally invasive methods and biomarkers may be suggested as alternative tools in diagnosing and monitoring eosinophilic esophagitis. While there is no consensus on the clinical usefulness of these biomarkers, our critical evaluation may suggest that the eosinophilic degranulation ratio (DGE/DGE + NDGE: degranulated eosinophils/degranulated eosinophils and non-degranulated eosinophils) in the esophagus may be critical for evaluating properly these biomarkers. An increasing trend may culminate in the potential clinical use of these biomarkers evaluated not only with the peak eosinophilic count, but also with the degranulation score upon regulatory bodies' approval to monitor eosinophilic esophagitis in the future. We strongly advocate for the necessity to score the esophageal biopsies with both a peak eosinophilic count and a score of the degranulated eosinophils.

Expanded population of low-density neutrophils in juvenile idiopathic arthritis.

Juvenile idiopathic arthritis (JIA), a clinically variable disease characterized by autoimmune arthritis, affects children, and its immunopathology remains elusive. Alterations in neutrophil biology play an important role in this disease. In the present study, we aimed to explore the features of low-density neutrophils (LDNs) in patients with JIA. Gene expression of peripheral blood mononuclear cells (PBMCs) from children with distinct subtypes of JIA was analyzed by NanoString Immunology panel. Presence of LDNs was ascertained by flow cytometry and the release of neutrophil-associated products were analyzed by LUMINEX. LDNs were detected in patients' peripheral blood mononuclear cells (PBMCs) after density gradient centrifugation. Transcriptomic analysis of JIA PBMCs revealed that genes related to neutrophil degranulation were markedly upregulated. The number of LDNs and level of their degranulation products increased in patients' PBMCs and correlated with serum calprotectin, but not with disease activity, sedimentation rate and C-reactive protein (CRP) levels. The phenotypes of LDNs varied from those of normal-density neutrophils and healthy donor LDNs. Phenotypical analysis revealed LDNs are immature and primed population with decreased suppressive capacity. A negative correlation between surface proteins CD62L, CD66b, and CD11b and the number of inflamed joints/JADAS was established. Our results describe LDNs as primed, degranulated, immature cells with impaired suppressive activities. This work thus contributes to the increasing body of evidence that LDNs in JIA are altered and their role in the disease immunopathogenesis and possible clinical associations should be investigated further.

Association of EDN levels in patients with asthma and correlation with FEV1%: A meta-analysis.

Background: Eosinophil-derived neurotoxin (EDN), an eosinophil degranulation product, is a good biomarker for eosinophilic inflammation of the airway. Several articles have shown that EDN levels are higher in patients with asthma than in controls, and EDN levels are correlated with the percentage of predicted forced expiratory volume in the first second (FEV1%) in patients with asthma. Their results were inconclusive. Methods: A comprehensive meta-analysis was performed to assess EDN levels between patients with asthma and controls, and the correlations between EDN and FEV1% in the patients with asthma. Fourteen relevant articles were identified from electronic data bases. Pooled standardized mean difference (SMD) with a 95% confidence interval (CI) for the difference of EDN levels between the patients with asthma and controls, and pooled coefficient (r) values with 95% CI for the correlations between EDN and FEV1%, respectively, were calculated. Results: A total of 14 articles were selected. Among the included reports, six articles related to the difference and eight essays on the correlation. Pooled effect size showed that EDN levels were higher in patients with asthma than in controls (SMD 2.85 [95% CI, 1.92-3.78]). Furthermore, the pooled effect size showed that EDN levels were negatively correlated with FEV1% in patients with asthma (r -0.21 [95% CI, -0.28 to -0.14]). Conclusion: EDN levels increased in the patients with asthma compared with in the controls. They were correlated with FEV1% in the patients with asthma, which indicated that EDN could be a reliable marker to monitor asthma's therapeutic effects.

Immunomodulation of neutrophils and platelets by TNF blockage in patients with juvenile idiopathic arthritis

Juvenile idiopathic arthritis (JIA) is a multifactorial autoimmune disease mediated by both adaptive and innate immunity. The role of neutrophils in the pathogenesis of autoimmune diseases is well-established; however, in JIA they are still markedly understudied. Here, we explored the neutrophil features and role of platelet-neutrophil aggregates in JIA patients and assessed the effect of TNF inhibitor (TNFi) therapy. We provide evidence of dysbalanced neutrophil subsets in JIA patients, with a shift towards immature and suppressive subpopulations that lack the cell-adhesion molecules. Correspondingly, patient sera contained high amounts of neutrophil- and platelet-related products. Transcriptomic analysis revealed neutrophil degranulation as the most affected process by TNFi therapy, which was mirrored by the decrease in degranulation products in the patient sera. Toll-like receptors −4, −7, and − 8 signaling pathways are particularly hyperresponsive in patients, but are strongly suppressed by TNFi. Overall, our study demonstrates augmented neutrophil and platelet responses in JIA patients.

Neutrophil Extracellular Vesicles and Airway Smooth Muscle Proliferation in the Natural Model of Severe Asthma in Horses.

Extracellular vesicles (EVs) contribute to intercellular communication through the transfer of their rich cargo to recipient cells. The EVs produced by LPS-stimulated neutrophils from healthy humans and horses increase airway smooth muscle (ASM) proliferation, but the roles of neutrophil EVs in asthma are largely unexplored. The aim of this study was to determine whether neutrophil-derived EVs isolated during the remission or exacerbation of asthma influence ASM proliferation differentially. Peripheral blood neutrophils were collected during remission and exacerbation in eight horses affected by severe asthma. The cells were cultured (±LPS), and their EVs were isolated by ultracentrifugation and characterized by laser scattering microscopy and proteomic analysis. The proliferation of ASM co-incubated with EVs was monitored in real time by electrical impedance. Two proteins were significantly upregulated during disease exacerbation in neutrophil EVs (MAST4 and Lrch4), while LPS stimulation greatly altered the proteomic profile. Those changes involved the upregulation of neutrophil degranulation products, including proteases known to induce myocyte proliferation. In agreement with the proteomic results, EVs from LPS-stimulated neutrophils increased ASM proliferation, without an effect of the disease status. The inhalation of environmental LPS could contribute to asthma pathogenesis by activating neutrophils and leading to ASM hyperplasia.

Lung Adenocarcinoma Cell Sensitivity to Chemotherapies: A Spotlight on Lipid Droplets and SREBF1 Gene.

Simple SummaryThe accumulation of lipid droplets (LDs) and the high expression of genes involved in LD formation, such as SREBF1 (sterol regulatory element binding transcription factor 1), are attributed to cancer cell resistance against anticancer drugs and poor prognosis. We assessed lung cancer cells with and without LDs for their sensitivity to chemotherapeutics cisplatin and etoposide. In either serum-free basal medium or inflammatory supernatants generated during neutrophil degranulation in vitro, both drugs strongly reduced SREBF1 expression, which did not parallel with LD formation and cell sensitivity to chemotherapeutics. Nevertheless, under basal conditions, SREBF1 expression in cancer cells correlated with LD levels, and the lower expression of SREBF1 in tumors than in adjacent nontumor tissues showed a prognostic value for overall better survival of patients with non-small-cell lung cancer. Strategies targeting lipid metabolism in cancer are promising therapeutic and/or diagnostic approaches.To explore the relationship between cancer cell SREBF1 expression, lipid droplets (LDs) formation, and the sensitivity to chemotherapies, we cultured lung adenocarcinoma cells H1299 (with LD) and H1563 (without LD) in a serum-free basal medium (BM) or neutrophil degranulation products containing medium (NDM), and tested cell responses to cisplatin and etoposide. By using the DESeq2 Bioconductor package, we detected 674 differentially expressed genes (DEGs) associated with NDM/BM differences between two cell lines, many of these genes were associated with the regulation of sterol and cholesterol biosynthesis processes. Specifically, SREBF1 markedly declined in both cell lines cultured in NDM or when treated with chemotherapeutics. Despite the latter, H1563 exhibited LD formation and resistance to etoposide, but not to cisplatin. Although H1299 cells preserved LDs, these cells were similarly sensitive to both drugs. In a cohort of 292 patients with non-small-cell lung cancer, a lower SREBF1 expression in tumors than in adjacent nontumor tissue correlated with overall better survival, specifically in patients with adenocarcinoma at stage I. Our findings imply that a direct correlation between SREBF1 and LD accumulation can be lost due to the changes in cancer cell environment and/or chemotherapy. The role of LDs in lung cancer development and response to therapies remains to be examined in more detail.

Influence of Malignant Pleural Fluid from Lung Adenocarcinoma Patients on Neutrophil Response.

Simple SummaryThis study provides novel information about the role of neutrophils in malignant pleural effusion (MPE) and hallmarks their clinical relevance. Since these cells have emerged as important regulators of cancer, we characterized their phenotype and functions in MPE microenvironment. We found that neutrophil-derived products (degranulation molecules and neutrophil extracellular traps (NETs)) were increased in MPE. In addition, NETs were associated with a worse outcome in lung adenocarcinoma patients with MPE.Malignant pleural effusion (MPE) is a common severe complication of advanced lung adenocarcinoma (LAC). Neutrophils, an essential component of tumor infiltrates, contribute to tumor progression and their counts in MPE have been associated with worse outcome in LAC. This study aimed to evaluate phenotypical and functional changes of neutrophils induced by MPE to determine the influence of MPE immunomodulatory factors in neutrophil response and to find a possible association between neutrophil functions and clinical outcomes. Pleural fluid samples were collected from 47 LAC and 25 heart failure (HF) patients. We measured neutrophil degranulation products by ELISA, oxidative burst capacity and apoptosis by flow cytometry, and NETosis by fluorescence. The concentration of degranulation products was higher in MPE-LAC than in PE-HF. Functionally, neutrophils cultured with MPE-LAC had enhanced survival and neutrophil extracellular trap (NET) formation but had reduced oxidative burst capacity. In MPE, NETosis was positively associated with MMP-9, P-selectin, and sPD-L1 and clinically related to a worse outcome. This is the first study associating NETs with a worse outcome in MPE. Neutrophils likely contribute to tumor progression through the release of NETs, suggesting that they are a potential therapeutic target in LAC.

Prospects and Applications of Natural Blood-Derived Products in Regenerative Medicine.

Currently, there are a number of therapeutic schemes used for the treatment of various types of musculoskeletal disorders. However, despite the use of new treatment options, therapeutic failure remains common due to impaired and delayed healing, or implant rejection. Faced with this challenge, in recent years regenerative medicine started looking for alternative solutions that could additionally support tissue regeneration. This review aims to outline the functions and possible clinical applications of, and future hopes associated with, using autologous or heterologous products such as antimicrobial peptides (AMPs), microvesicles (MVs), and neutrophil degranulation products (DGP) obtained from circulating neutrophils. Moreover, different interactions between neutrophils and platelets are described. Certain products released from neutrophils are critical for interactions between different immune cells to ensure adequate tissue repair. By acting directly and indirectly on host cells, these neutrophil-derived products can modulate the body’s inflammatory responses in various ways. The development of new formulations based on these products and their clinically proven success would give hope for significant progress in regenerative therapy in human and veterinary medicine.

The Effect of Neutrophil-Derived Products on the Function of Leukocytes Obtained after Titanium Implantation in the Ovine Model.

Simple SummaryTitanium is one of the most commonly used biomaterials for implantation as a part of the orthopedic procedures. However, this biomaterial can cause an excessive inflammatory response, even leading to rejection of the implant. Therefore, the aim of our study was to assess the overall organism response after insertion of Ti implant and the activity of neutrophils and monocyte-derived macrophages (MDM), to evaluate the possible negative effect of this biomaterial on the host cells. Our study revealed that insertion of the Ti implant did not evoke systemic inflammatory response or activation of leukocytes. Additionally, we evaluated the activity of neutrophils and MDM after stimulation with autologous neutrophil products, namely, antimicrobial neutrophil extract and neutrophil degranulation product as two potential regulators of inflammatory response. Antimicrobial neutrophil extract appeared to be a factor causing the decrease of secretory neutrophil response and polarization of MDM towards pro-resolving phenotype, whereas the neutrophil degranulation product acted as pro-inflammatory.Titanium (Ti) is currently the most common biomaterial used for orthopedic implants; however, these implants may cause deleterious immune response. To investigate the possible mechanisms involved in excessive inflammation, we assessed the activity of neutrophils and monocyte-derived macrophages (MDMs) during the insertion of the Ti implant in a sheep model. The study was conducted on 12 sheep, 4 of which were control animals and 8 were in the experimental group with inserted Ti implant. Neutrophil secretory response was estimated at two time points T0 before surgery and T1 1 h after implantation and was based on the release of enzymes from neutrophil granules and reactive oxygen and nitrogen species (RONS) generation. MDM function was evaluated 5 months after implantation, on the basis of RONS generation arginase activity and morphological changes. Moreover, the influence of some autologous neutrophil derived products, namely, antimicrobial neutrophil extract (ANE) and neutrophil degranulation products (DGP) on leukocytes was estimated. Our study revealed that Ti implant insertion did not cause any adverse effects up to 5 months after surgical procedure. Stimulation of neutrophil cultures with ANE decreased the enzyme release as well as superoxide generation. Treatment of MDM with ANE diminished superoxide and NO generation and increased arginase activity. On the other hand, MDM stimulated with DGP showed elevated superoxide and NO generation as well as decreased arginase activity. To summarize, ANE exerted an anti-inflammatory and pro-resolving effect on studied leukocytes, whereas DGP acted as pro-inflammatory.

Dairy Processing Affects the Gut Digestion and Microecology by Changing the Structure and Composition of Milk Fat Globules.

Milk fat globules (MFGs) are the major source of energy for infants' dietary intake. In this study, the effects of changes in the structure and composition of MFG after dairy processing on lipolysis and immune regulation were investigated. Pasteurized MFG tends to form protein aggregates to prevent lipolysis. However, the aggregate is rich in neutrophil degranulation products, which are effective in killing pathogens. Homogenized MFG has the lowest hydrolysis rate due to the reconstituted anti-lipase barrier and exposed apolipoprotein. Simultaneously, the reconstituted barrier can compensate for the lack of the complement cascade. Spray-dried MFG had the highest hydrolysis rate attributable to the disrupted MFG barrier and the release of lipoprotein lipase and endothelial lipase. The immunomodulatory properties of spray-dried MFG proteins are mainly mediated by the toll-like receptor (TLR) signaling pathway. This research provides the improvement basis of dairy processing and functional infant formulas.

Oral dimethyl fumarate induces changes within the peripheral neutrophil compartment of patients with psoriasis that are linked with skin improvement.

Dimethyl fumarate (DMF) is a treatment for moderate-to-severe psoriasis and multiple sclerosis. DMF therapy typically improves skin inflammation within the first 3months of treatment. DMF is a prodrug that generates the hydroxycarboxylic acid receptor 2 (HCA2) agonist, monomethyl fumarate (MMF). Despite widespread clinical use, DMF's mechanism of action is not fully understood. We wished to characterize the changes induced by DMF in peripheral neutrophils within the first 3months of treatment to better understand its early antipsoriatic effects. Flow cytometry was used to assess T-cell and neutrophil frequencies, apoptosis and activation phenotype. In vitro culture of neutrophils with DMF and MMF was used to evaluate apoptosis and HCA2 internalization. Serum levels of neutrophil degranulation products were measured by enzyme-linked immunosorbent assay. Patients with psoriasis had significantly higher leucocyte counts at baseline compared with controls, with a large population of pro-inflammatory CD62Llo CD11bbright neutrophils. Analysis revealed that DMF treatment reduced the frequency of CD62Llo CD11bbright neutrophils and serum levels of neutrophil activation markers. This reduction was not linked to increased apoptosis. Our results reveal a novel invivo effect of DMF therapy on pro-inflammatory neutrophils that likely contributes to this treatment's antipsoriatic efficacy.

Interactions Between Neutrophils and Periodontal Pathogens in Late-Onset Periodontitis.

Late-onset periodontitis is associated with a series of inflammatory reactions induced by periodontal pathogens, such as Porphyromonas gingivalis, a keystone pathogen involved in periodontitis. Neutrophils are the most abundant leukocytes in the periodontal pocket/gingival crevice and inflamed periodontal tissues. They form a “wall” between the dental plaque and the junctional epithelium, preventing microbial invasion. The balance between neutrophils and the microbial community is essential to periodontal homeostasis. Excessive activation of neutrophils in response to periodontal pathogens can induce tissue damage and lead to periodontitis persistence. Therefore, illuminating the interactions between neutrophils and periodontal pathogens is critical for progress in the field of periodontitis. The present review aimed to summarize the interactions between neutrophils and periodontal pathogens in late-onset periodontitis, including neutrophil recruitment, neutrophil mechanisms to clear the pathogens, and pathogen strategies to evade neutrophil-mediated elimination of bacteria. The recruitment is a multi-step process, including tethering and rolling, adhesion, crawling, and transmigration. Neutrophils clear the pathogens mainly by phagocytosis, respiratory burst responses, degranulation, and neutrophil extracellular trap (NET) formation. The mechanisms that pathogens activate to evade neutrophil-mediated killing include impairing neutrophil recruitment, preventing phagocytosis, uncoupling killing from inflammation, and resistance to ROS, degranulation products, and NETs.

Eosinophil-derived neurotoxin as a biomarker in children with allergic rhinitis

Eosinophil-derived neurotoxin (EDN), an eosinophil degranulation product, is considered as a biomarker for allergic diseases, such as asthma. The purpose of our study was to investigate whether EDN reflects eosinophilic inflammation in children with allergic rhinitis.

Application of Natural Neutrophil Products for Stimulation of Monocyte-Derived Macrophages Obtained before and after Osteochondral or Bone Injury.

We evaluated the use of some neutrophil products, namely; autologous rabbit antimicrobial neutrophil extract (rANE), heterologous porcine antimicrobial neutrophil extract (pANE), neutrophil degranulation products (DGP) and neutrophil microvesicles (MVs) for stimulation of monocyte-derived macrophages (MDMs) to improve healing. Two animal models were evaluated; the rabbit model for autologous osteochondral transplantation (OT) with application of rabbit ANE, DGP or MVs for MDMs stimulation, and the ovine model of the insertion of a Ti implant with the use of porcine ANE, and ovine DGP or MVs for MDMs stimulation. Macrophage activity was assessed on the basis of free radical generation and arginase activity. We estimated that DGP acted in a pro-inflammatory way both on rabbit and ovine MDMs. On the other hand, MVs acted as anti-inflammatory stimulator on MDMs in both experiments. The response to ANE depended on origin of extract (autologous or heterologous). Macrophages from rabbits before and after OT stimulated with autologous extract generated lower amount of NO and superoxide, especially after transplantation. In the ovine model of Ti implant insertion, heterologous ANE evoked increased macrophage pro-inflammatory activity. Our study revealed that these neutrophil products could regulate activity of macrophages, polarizing them into pro-or anti-inflammatory phenotypes that could enhance bone and osteochondral tissue healing.

Image Analysis of Eosinophil Peroxidase Immunohistochemistry for Diagnosis of Eosinophilic Esophagitis

Diagnosis of eosinophilic esophagitis (EoE) requires manual quantification of tissue eosinophils. Eosinophil peroxidase (EPX) is an eosinophil-specific, cytoplasmic granule protein released during degranulation. The objective of this study was to evaluate image analysis of EPX immunohistochemistry as an automated method for histologic diagnosis of EoE. We performed a secondary analysis of prospectively collected esophageal biopsies obtained from adult subjects with EoE and controls. Tissue sections were stained with hematoxylin and eosin (H&E) and evaluated for peak eosinophils per high power field (eos/hpf). The same slides were de-stained and re-stained to detect EPX for direct comparison. Slides were digitized, and EPX staining area/mm2 was quantified using image analysis. Paired samples were compared for changes in EPX staining in treatment responders and non-responders. Thirty-eight EoE cases and 49 controls were analyzed. Among EoE subjects, matched post-treatment biopsies were available for 21 responders and 10 non-responders. Baseline EPX/mm2 was significantly increased in EoE subjects and decreased in treatment responders. EPX quantification correlated strongly with eos/hpf (r = 0.84, p < 0.0001) and identified EoE subjects with high diagnostic accuracy (AUC 0.95, p < 0.0001). The optimal diagnostic EPX-positive pixel/area threshold was 17,379 EPX/mm2. Several controls (5/49) with < 15 eos/hpf on H&E staining exceeded this cutoff. EPX/mm2 correlates strongly with eos/hpf, accurately identifies subjects with EoE, and decreases in treatment responders. Automated quantification of intact eosinophils and their degranulation products may enhance pathologic assessment. Future studies are needed to correlate EPX/mm2 with symptoms, endoscopic findings, and esophageal distensibility.

Resistin is associated with overall survival in non-small cell lung cancer patients during nivolumab treatment.

Sincethe role of resistin was evaluated onlyin patients with non-small cell lung cancer (NSCLC) not treated with immunotherapy, we aimed to evaluatelevels of resistin during immunotherapy (nivolumab) and its prognostic role with regard to OS. From a cohort of 78 patients with advanced NSCLC enrolled in a prospective study at Ospedale Policlinico San Martino in Genoa (Italy), 43 patients have been considered for this sub-analysis because of the availability of samples. Before and during nivolumab administration, clinical information and blood samples were collected and resistin, matrix metalloproteinase (MMP)-8, MMP-9, and myeloperoxidase were evaluated by enzyme-linked immunosorbent assay (ELISA). Median age was 71 with a prevalence of males and former smokers. Median resistin levels presented a peak at cycle 2 and then dropped down until the last cycle. Resistin correlated with all neutrophil degranulation products at cycle 1 (except for MMP-9) and at cycle 2 as well as with white blood cells and neutrophils. By a ROC curve analysis, a resistin value at cycle 2 of 19ng/mL was tested as the best cut-off point for OS. Kaplan-Meier analysis demonstrated that patients above the resistin cut-off experienced a reduced OS (median OS 242.5 vs. 470days, p = 0.0073), as confirmed by Cox proportional hazards regression analysis. Resistin levels > 19ng/mL at the time of the second cycle of nivolumab treatment independently predict a reduced OS in patients with advanced NSCLC.

Poor Correlation of Oral Swabs with Esophageal Eosinophil Counts.

Eosinophilic esophagitis (EoE) is a chronic condition that requires repeated endoscopies/biopsies to track the disease and treatment response. This invasive procedure involves risk to the patient and has significant costs. We studied whether the detection of specific proteins (cytokines and eosinophil degranulation products) from oral swabs could serve as a minimally invasive test for EoE. Swabs of the oral cavity (buccal and oropharyngeal) were obtained prior to endoscopy/biopsies in patients with EoE, possible EoE, and non-EoE patients in addition to obtaining additional esophageal biopsy tissue. ELISAs measuring the levels of cytokines IL-5, IL-8, IL-13, and eosinophil degranulation products including major basic protein (MBP), eosinophil derived neurotoxin (EDN), and eosinophil peroxidase (EPO) were performed on the samples. Comparisons were made to peak esophageal eosinophil counts. Tolerability of the swabs was evaluated. 43 patients, 4-17years old, participated in the study. Swabs were well tolerated and all showed measurable protein. 26 patients had EoE [14 active (> 15 eosinophils/high power field), 12 non-active], 17 patients did not have EoE. Results obtained from oral swabs showed poor correlation with those from esophageal tissue. Only measurement of eosinophil degranulation products EDN and EPO from esophageal tissues showed strong correlations with eosinophil counts. In this study, the levels of cytokines and eosinophil degranulation products detected from oral swabs did not correlate with esophageal eosinophilia, and their detection would be insufficient to displace endoscopy/biopsies.

Early post-operative vasoplegia in lung transplantation

Introduction Lung transplantation (LTx) is the main treatment for end-stage respiratory failure. The post-operative development of pulmonary and systemic inflammation represents a severe limitation leading to Primary Graft Dysfunction (PGD) and multi-organ failure with great impact on survival and clinical outcomes. We hypothesise that LTx is associated with a high incidence of vasoplegia and that this is associated with a profound inflammatory response and high mortality and organ dysfunction. To test our hypotheses, we have identified patients who developed vasoplegia in the first 48 hours after LTx, described the impact of the vasoplegia on the clinical impact and explored the role of neutrophil degranulation products, such as Myeloperoxidase (MPO) and Heparin Binding Protein (HBP). Methods We recruited 40 patients who underwent LTx in 2013-15 at Harefield Hospital. Clinical data were recorded. Vasoplegia was defined according to Tsiouris et al (1). PGD was diagnosed and scored according to ISHLT criteria 2005, Acute Kidney Injury (AKI) to KDIGO criteria. Blood samples were collected before surgery and at different time-points after the arrival in ITU and analysed for MPO and HBP levels (Axis-Shield Heparin Binding Protein EIA, Abcam MPO ELISA kit). Data were analysed and reported as appropriate. Results 13 patients developed post-operative vasoplegia (32%). Within this group, 6 patients also developed PGD 3 (46%, p 0.284) and 9 AKI (69%, p 0.312), 7 requiring haemofiltration (54%, p 0.032). Vasoplegic patients showed lower P/F ratios on day 1 (23.1 [17.3, 35.7] vs. 31.4 [24.4, 41.4], p 0.151), day 2 936.0 [29.0, 44.6] vs. 46.7 [41.4, 50.5], p 0.011), and day 3 939.6 [29.7, 46.9] vs. 48.3 [43.4, 53.2], p 0.004). They also had longer ventilation duration (159 [59, 971] hours vs. 29 [15, 114] hours, p 0.012) and longer ITU stay (22 [5, 43] days vs. 5 [3, 9] days, p 0.029). Overall survival was not significantly affected by vasoplegia (log rank 1.005 p 0.316). Vasoplegic patients exhibit a tendency for higher levels of MPO early after surgery (232 [140, 406] ng/ml vs. 170 [110, 336] ng/ml before surgery (p 0.247), 566 [84, 1246] ng/ml vs. 312 [121, 658] ng/ml at the arrival in ITU (p 0.504), 441 [231, 753] ng/ml vs. 300 [142, 518] ng/ml at 6 hours (p 0.353), 160 [52, 447] ng/ml vs. 139 [112, 744] ng/ml at 12 hours (p 0.200)), but similar levels of HBP. Discussion These results identify high incidence of vasoplegia in the LTx population. One third of our patients developed vasoplegia in the early post-operative period with significant impact on the early post-operative function in terms of gas exchange and need for ventilatory support and level 3 care. Moreover, the data may suggest potential involvement of neutrophil-derived myeloperoxidase and, therefore oxidative stress, in the early postoperative period events.

Human Toxocara Infection: Allergy and Immune Responses.

Toxocariasis is a cosmopolitan infection that occurs in various regions worldwide, more frequently in developing countries. Chronic infections with Toxocara species in humans are associated with the production of high levels of specific and non-specific antibodies of all isotypes and IgG subclasses and a cytokine response characterized by the production of Th2 cytokines including IL-4, IL-5 and IL-13 by Peripheral Blood Monocytes (PBMCs) and Leukocytes (PBLs) in whole blood cultures. Other Th2 effector responses are also prominent during infection, reflected by elevated numbers of peripheral blood eosinophils and increased expression of eosinophil degranulation products. The production of IFN-γ by PBMCs/PBLs stimulated with Toxocara-secreted proteins is not prominent in toxocariasis but IL-10 production may be increased in infected individuals. The relationship between Toxocara species with allergic reactions was reported in the recent century. Experimental and epidemiological investigations revealed that toxocariasis with this parasite led to the development of allergic symptoms, such as asthma. However, the findings are conflicting since in other investigations no association between these two immunopathologies has been reported. The present review endeavours to summarize the data on Toxocara species and findings from studies on the relationship of toxocariasis with symptoms and signs of allergy. Furthermore, the mechanisms of immune responses and the factors associated between allergy and Toxocara infection are discussed.