Eosinophils Research Articles

Reaction of eosinophilic granulocytes of guinea pigs airways to the experimental allergic inflammation according to the morphometric study

Background. After creating an animal model of experimental airways allergic inflammation using ovalbumin, scientists mainly studied the reaction of the cellular and humoral links of the acquired specific immunity. At the same time, it is known that the development of allergic inflammation of the airways to the effect of the environmental chemoattractants is primarily the result of the local innate immune system response. Pulmonary neuroendocrine cells of the airway epithelium with the help of neuropeptides activate the secretion of IL-5 and IL-13 by type 2 innate lymphoid cells IL-5 determines the eosinophilic inflammation. The question of the reaction of the components of innate immunity of the respiratory tract to the allergic inflammation in most aspects remains open. open. Objective. The purpose of the current study was to define the reaction of eosinophilic granulocytes in guinea pigs lungs as an important component of the innate immunity of the respiratory tract to experimental ovalbumin-induced allergic inflammation according to the data of a morphometric study. Methods. Histological, morphometric, statistical methods were used to study the topographic features and the number of eosinophils in the lungs of male 48 guinea pigs on the 23rd, 30th, 36th and 44th days after initiation of the experimental ovalbumin-induced allergic airway inflammation. Results. Our study demonstrates a significant reaction occurs on the part of the cellular link of innate immunity, which consists in the activation of eosinophils, in the experimental model of ovalbumin-induced airways allergic inflammation. The most significant changes were observed in the distal parts of the intrapulmonary airways and in the structures of the pulmonary acinus in the early period of the development of the inflammatory process (23rd and 30th days after the start of the experiment). The late (36th and 44th days after the start of the experiment) period of the development of an allergic inflammatory process in the lungs is accompanied by a gradual decrease in the activity of eosinophilic inflammation. At the same time, the quantitative indicators remain statistically significantly higher in the intact and control groups, which indicates the continuation of allergic inflammation in the absence of the action of the allergen and is a manifestation of the violation of the recovery and adaptation processes in the local immune system of the lung. Conclusion. After ovalbumin-sensitization and aeroallergization eosinophilic inflammation develops in the lungs of a guinea pig, as a result of the reaction of the innate immunity to the action of an allergen. Actively and first of all, allergic eosinophilic inflammation develops in the distal airways (terminal bronchioles) and in the connective tissue stroma of the pulmonary acinus.

Resident leukocyte populations and inflammatory response in the abdominal adipose tissue of the red seabream Pagrus major

Adipose tissue is not only an energy depot but also an important endocrine and immunological tissues in mammals. However, the study of functions of fish adipose tissue, other than lipid storage, is considerably limited. In the present study, we investigated resident leukocyte populations in the visceral adipose tissue (VAT) of the red sea bream, Pagrus major, an important species of Japanese aquaculture with a well-developed VAT. We found that the tissue contains a significant number of leukocytes; it is mainly composed of eosinophilic granulocytes, macrophages, unclassified large cells, and a smaller number of lymphocytes. The VAT cellular composition is similar to that of peritoneal cells, but not of the peripheral blood leukocytes. Gene expression analysis also confirmed that all examined genes, including CD4, CD8α, membrane and secretory IgM heavy chains, macrophage-colony stimulating factor receptors (type 1 and 2), macrophage mannose receptor, MHC class II, and Toll-like receptors 2, 3, 5, and 9, were expressed in the VAT. Intraperitoneal injection of formalin-killed Vibrio anguillarum markedly upregulated the mRNA expression of IL-1β, IL-8, and IL-6 in the VAT within six hours. In addition, in vitro bacterial stimulation upregulated these cytokines. These results indicate that the VAT of P. major is an immune-competent organ that evokes an inflammatory response against intraperitoneally introduced bacteria.

Eosinophils are dispensable for development of MOG35–55-induced experimental autoimmune encephalomyelitis in mice

Experimental autoimmune encephalomyelitis (EAE) represents the mouse model of multiple sclerosis, a devastating neurological disorder. EAE development and progression involves the infiltration of different immune cells into the brain and spinal cord. However, less is known about a potential role of eosinophil granulocytes for EAE disease pathogenesis. In the present study, we found enhanced eosinophil abundance accompanied by increased concentration of the eosinophil chemoattractant eotaxin-1 in the spinal cord in the course of EAE induced in C57BL/6 mice by immunization with MOG35–55 peptide. However, the absence of eosinophils did not affect neuroinflammation, demyelination and clinical development or severity of EAE, as assessed in ∆dblGATA1 eosinophil-deficient mice. Taken together, despite their enhanced abundance in the inflamed spinal cord during disease progression, eosinophils were dispensable for EAE development.

Evaluation of Blood Markers at Admission for Predicting Community Acquired Pneumonia in Chronic Obstructive Pulmonary Disease

Acute exacerbations of Chronic Obstructive Pulmonary Disease (AECOPD) and community acquired pneumonia (CAP) are two common acute attacks in COPD patients and it is not always easy to determine whether a COPD patient at admission has parenchymal infection or bronchial infection. Comprehensive comparison between AECOPD patients and CAP patients with COPD (COPD + CAP) can help us understand them better. We retrospectively collected the medical records of AECOPD and COPD + CAP patients. Systemic inflammation, eosinophilic inflammation, damage to other organs, common chronic comorbidities, structural changes, phenotype and endotype distributions and coagulation functions between two groups were compared and correlations of these characteristics in total subjects, AECOPD patients and COPD + CAP patients were analyzed. Logistic regression analysis was performed to select helpful biomarkers for distinguishing between them. Receiver operator characteristic (ROC) curve was plotted to assess the diagnostic value of selected biomarkers and their combination. A nomogram was established for the differential diagnosis of AECOPD and COPD + CAP. A total of 206 patients were included into our analysis. In these subjects, 104 patients were classified as AECOPD group and 102 patients were considered to have COPD + CAP mainly based on their chest CT scan results. The counts of eosinophils (EOS), basophils (BAS) and lymphocytes (LYM) and percentage of total white blood cell count, hemoglobin and hematocrit were increased in AECOPD patients compared with COPD + CAP patients. The counts of neutrophils (NEU) and percentage of total white blood cell count, C-reactive protein (CRP), Erythrocyte sedimentation rate (ESR), fibrinogen, D-dimer and N-Terminal pro-brain natriuretic peptide (NT-proBNP) levels were increased in COPD + CAP patients. After logistic regression analysis, EOS < 0.5 × 109/L, ESR ≥ 8 mm/H and NT-proBNP ≥ 100 pg/mL were selected as helpful biomarkers for diagnosis of COPD + CAP instead of AECOPD. Area under the ROC curve (AUC) of the combination of selected biomarkers was 0.764(0.698-0.829). A nomogram was established and the calibration curve suggested that fitting efficiency of the nomogram was good. AECOPD and COPD + CAP are markedly different, mainly reflected in eosinophilic inflammation, systemic inflammation and coagulation function. Correlations between some common inflammatory biomarkers are also different in the two groups. A nomogram was established to offer help to clinicians for differential diagnosis of these two diseases.

HPV-negative Squamous Cell Carcinomas of the Cervix With Special Focus on Intraepithelial Precursor Lesions.

Recently, the World Health Organization (WHO) recognized human papilloma virus (HPV)-independent invasive cervical squamous cell carcinoma (SCC) without recognizing the existence of precursor lesions. This is a detailed characterization of 3 preinvasive lesions and 6 invasive SCC negative for HPV-DNA (32 genotypes), HPV-mRNA (14 genotypes) and genomic HPV sequencing. We evaluated histologic features, expression of p16ink4a, p53, CK7, and CK17, aberrations in 50 cancer genes and chromosomal copy number variations. HPV-negative preinvasive lesions were extensive basaloid or highly differentiated keratinizing intraepithelial proliferations of 3 to 20 cell layers thickness, partly with prominent cervical gland involvement. Overall, 2/3 intraepithelial lesions and the in situ component of 1/6 SCC showed p16ink4a block staining, while 1/6 in situ component revealed heterogenous p16ink4a staining. All invasive components of keratinizing SCC were p16ink4a-negative. Preinvasive and invasive SCC showed inconsistent CK7 and CK17 staining. Nuclear p53 overexpression was restricted to the TP53 gene mutated SCC. The highly vascularized peritumoral stroma showed a dense inflammatory infiltrate including plasma cells and intratumoral and peritumoral eosinophilic granulocytes. Inconsistent somatic gene mutations (PIK3CA, STK11, TP53, SMARC2B, and GNAS) occurred predominantly in nonhotspot locations at low mutational frequency in 3/6 SCC. Consistent aberrations included the pathogenic (angiogenic) germline polymorphism Q472H in the KDR gene (7/9 patients), and chromosome 3q gains (4/9 patients). In conclusion, HPV-negative intraepithelial cervical precancerous lesions exist, either as highly differentiated keratinized intraepithelial proliferations reminiscent of differentiated vulvar intraepithelial neoplasia, or undifferentiated basaloid intraepithelial lesions with occasional p16ink4a block staining resembling high-grade squamous intraepithelial lesion. Gains of chromosome 3q, angiogenic germline variants the inflammatory infiltrate may contribute to progression of HPV-negative cervical carcinogenesis.

Interleukin-5-induced eosinophil population improves cardiac function after myocardial infarction.

Interleukin (IL)-5 mediates the development of eosinophils (EOS) that are essential for tissue post-injury repair. It remains unknown whether IL-5 plays a role in heart repair after myocardial infarction (MI). This study aims to test whether IL-5-induced EOS population promotes the healing and repair process post-MI and to reveal the underlying mechanisms. MI was induced by permanent ligation of the left anterior descending coronary artery in wild-type C57BL/6 mice. Western blot and real-time polymerase chain reaction revealed elevated expression of IL-5 in the heart at 5 days post-MI. Immunohistostaining indicated that IL-5 was secreted mainly from macrophages and CD127+ cells in the setting of experimental MI. External supply of recombinant mouse IL-5 (20 min, 1 day, and 2 days after MI surgery) reduced the infarct size and increased ejection fraction and angiogenesis in the border zone. A significant expansion of EOS was detected in both the peripheral blood and infarcted myocardium after IL-5 administration. Pharmacological depletion of EOS by TRFK5 pretreatment muted the beneficial effects of IL-5 in MI mice. Mechanistic studies demonstrated that IL-5 increased the accumulation of CD206+ macrophages in infarcted myocardium at 7 days post-MI. In vitro co-culture experiments showed that EOS shifted bone marrow-derived macrophage polarization towards the CD206+ phenotypes. This activity of EOS was abolished by IL-4 neutralizing antibody, but not IL-10 or IL-13 neutralization. Western blot analyses demonstrated that EOS promoted the macrophage downstream signal transducer and activator of transcription 6 (STAT6) phosphorylation. IL-5 facilitates the recovery of cardiac dysfunction post-MI by promoting EOS accumulation and subsequent CD206+ macrophage polarization via the IL-4/STAT6 axis.

Eosinophils Correlate with Epithelial-Mesenchymal Transition in Chronic Rhinosinusitis with Nasal Polyps

Introduction: Chronic inflammation and tissue remodeling always occur together in chronic rhinosinusitis (CRS). Epithelial-mesenchymal transition (EMT) plays a critical role in airway remodeling. Objective: Changes of epithelial cells in sinus mucosa in different subtypes of CRS, especially in eosinophilic chronic rhinosinusitis with nasal polyps, and the role of EMT and eosinophils (EOS) in airway remodeling are still unknown. Methods: We included 85 patients in this study. They were divided into 4 groups: a normal control (NC) group, a chronic rhinosinusitis without nasal polyps (CRSsNP) group, an eosinophilic chronic rhinosinusitis with nasal polyps (ECRSwNP) group, and a noneosinophilic chronic rhinosinusitis with nasal polyps (non-ECRSwNP) group. Clinical data were all collected and analyzed. Standard hematoxylin and eosin staining, immunohistochemical staining, and 2-color immunofluorescence staining were performed. Biomarkers of EMT, epithelial cadherin, and vimentin were labeled. The immunohistochemistry results of each group were counted and statistically analyzed. Results and Conclusion: E-cadherin was downregulated, and vimentin was upregulated in epithelial tissue from the ECRSwNP group, compared with that from the control group and the other groups. The number of vimentin-expressing epithelial cells correlated with sinus CT imaging Lund-Mackay scores (r = 0.560, p < 0.001). Moreover, expression levels of vimentin in the epithelium were associated with numbers of infiltrating EOS in tissues (r = 0.710, p < 0.001) and the peripheral blood EOS ratio (r = 0.594, p < 0.001). EMT occurred in patients with CRSwNP, especially in those with ECRSwNP. Epithelial reprogramming correlates with eosinophil infiltration and disease severity. Eosinophils contributed to impairment of epithelial function and promoted EMT in CRSwNP.

Comparing Patient Characteristics, Clinical Outcomes, and Biomarkers of Severe Asthma Patients in Taiwan.

Purpose: To understand the association between biomarkers and exacerbations of severe asthma in adult patients in Taiwan. Materials and Methods: Demographic, clinical characteristics and biomarkers were retrospectively collected from the medical charts of severe asthma patients in six hospitals in Taiwan. Exacerbations were defined as those requiring asthma-specific emergency department visits/hospitalizations, or systemic steroids. Enrolled patients were divided into: (1) those with no exacerbations (non-exacerbators) and (2) those with one or more exacerbations (exacerbators). Receiver operating characteristic curves were used to determine the optimal cut-off value for biomarkers. Generalized linear models evaluated the association between exacerbation and biomarkers. Results: 132 patients were enrolled in the study with 80 non-exacerbators and 52 exacerbators. There was no significant difference in demographic and clinical characteristics between the two groups. Exacerbators had significantly higher eosinophils (EOS) counts (367.8 ± 357.18 vs. 210.05 ± 175.24, p = 0.0043) compared to non-exacerbators. The optimal cut-off values were 292 for EOS counts and 19 for the Fractional exhaled Nitric Oxide (FeNO) measure. Patients with an EOS count ≥ 300 (RR = 1.88; 95% CI, 1.26–2.81; p = 0.002) or FeNO measure ≥ 20 (RR = 2.10; 95% CI, 1.05–4.18; p = 0.0356) had a significantly higher risk of exacerbation. Moreover, patients with both an EOS count ≥ 300 and FeNO measure ≥ 20 had a significantly higher risk of exacerbation than those with lower EOS count or lower FeNO measure (RR = 2.16; 95% CI, 1.47–3.18; p = < 0.0001). Conclusions: Higher EOS counts and FeNO measures were associated with increased risk of exacerbation. These biomarkers may help physicians identify patients at risk of exacerbations and personalize treatment for asthma patients.

Leucocytes, urea and glucose levels in Albino Wistar rats exposed to doses of isolated Achatina achatina snail lectin

There are five (5) types of mature White blood cells (WBC) or Leucocytes found in the peripheral blood viz, Neutrophils (NEU), Eosinophils (EOS) and Basophils (BAS) (granulocytes); Monocytes (MON) and Lymphocytes (LYM) (agranulocytes). Urea is an organic chemical compound, and is essentially the waste produced by the body after metabolizing protein. Urea levels can be used to detect diseases and disorders that affect the kidneys. A common disease related to irregular management of glucose is diabetes. Lectins are proteins that recognize specifically and bind reversibly the carbohydrate-containing molecules of foreign cells and that elicit diverse physiological responses in various organisms. A total of 120 samples of Nigeria Achatina achatina snail specie were collected, authenticated at the Zoology Department of University of Nigeria, Nsukka and 80mls of pooled crude Lectin extract was obtained. Purifications were performed on 20mls of the crude extract in three steps viz, Ammonium sulphate precipitation and Dialysis (Partial purifications), Con A Sepharose 4B affinity Chromatography column (Complete purification). The affinity purified Lectin was used for all the tests conducted in this research. The crude, partially and complete/affinity purified Lectin extracts were subjected to Haemagglutination tests. The Lectin was further assessed to determine its effects on Leucocytes, Urea and Glucose as follows: A total of Thirty-five (35) male Albino Wistar Rats weighing 101-180g and aged 2-3 months obtained from the Animal house of University of Nigeria, Nsukka, were used in this research. The animals were Grouped into 5 (A-E) and allowed for 2 weeks acclimatization. Graded doses of 0.04ml, 0.05ml and 0.06ml of the Affinity purified Lectin were injected intra-peritoneally into each of the Rats in Groups A-D (test groups) according to their body weights at intervals of 2 days for 1 week. Group E served as the control. Two (2) mls of blood was collected from each of the Rats before and 24 hours after the last day of Lectin Doses injections for the following tests: WBC-Total and Differential counts (using Sysmex Corporation, 1999 automated equipment), Urea and Glucose estimations (performed by means of Urease-Berthelot and GOD-PAP Randox Monza automated analyser methods respectively). The results of the research showed as follows: On complete/affinity purification, 15mls of pure sample containing only the high molecular weight Lectin was obtained. The haemagglutination tests conducted showed on standardization preferential agglutination with Blood group A type. Bar Charts statistics show that there was Post Lectin Doses injections mean increase in Total WBC, NEU, LYM and decrease in MON, EOS, BAS, Urea and Glucose levels. However, the differences in Pre and Post Lectin Doses injections mean values of these parameters were further subjected to One way analysis of variance (ANOVA) test statistics to determine if statistically significant. The ANOVA statistics show that the effects of the Lectin on all the assessed Leucocytes parameters viz, Total WBC, and Differential LYM, NEU, MON, EOS, BAS, the Urea and Glucose levels were found to be statistically insignificant. However, the EOS values of only group A was statistically significant. This research has therefore succeeded in Assessment of Activities of the A. achatina snail Lectin on Leucocytes, Glucose and Urea levels.

A rare case of multicentric eosinophilic granulocytic sarcoma as cause of hindlimb weakness and non‐regenerative anaemia in a minipig

AbstractA 6‐year‐old male, neutered minipig developed progressive hindlimb weakness and finally became recumbent. Clinical and laboratory examination revealed restricted hindlimb mobility and weight bearing with unimpaired reflexes and proprioception as well as marked non‐regenerative macrocytic anaemia, neutropenia and lymphocytopenia. Underlying bone marrow disease and haematopoietic neoplasia were assumed after exclusion of spinal disc herniation and infectious causes for anaemia. Despite stabilising therapy, the animal died 7 days later. Postmortem examination revealed multiple beige‐coloured nodules affecting many organs including the bone marrow and consisting of neoplastic round cells with intracytoplasmic eosinophilic granules. Using various histopathological stains and immunohistochemistry, the neoplastic cells were determined to be of eosinophil granulocyte origin. Granulocytic sarcoma is a rare form of myeloid neoplasia with only two published porcine cases. With the increasing popularity of minipigs as companion animals, extensive awareness and information concerning illnesses in these animals should be available to practitioners.

Blood eosinophil counts in the general population and airways disease: a comprehensive review and meta-analysis.

BackgroundThe clinical context for using blood eosinophil (EOS) counts as treatment–response biomarkers in asthma and COPD requires better understanding of EOS distributions and ranges. We describe EOS distributions and ranges published in asthma, COPD, control (non-asthma/COPD) and general populations.MethodsWe conducted a comprehensive literature review and meta-analysis of observational studies (January 2008 to November 2018) that included EOS counts in asthma, severe asthma, COPD, control and general populations. Excluded studies had total sample sizes <200, EOS as inclusion criterion, hospitalised population only and exclusively paediatric participants.ResultsOverall, 91 eligible studies were identified, most had total-population-level data available: asthma (39 studies), severe asthma (12 studies), COPD (23 studies), control (seven studies) and general populations (14 studies); some articles reported data for multiple populations. Reported EOS distributions were right-skewed (seven studies). Reported median EOS counts ranged from 157–280 cells·µL−1 (asthma, 22 studies); 200–400 cells·µL−1 (severe asthma, eight studies); 150–183 cells·µL−1 (COPD, six studies); and 100–160 cells·µL−1 (controls, three studies); and 100–200 cells·µL−1 (general populations, six studies). The meta-analysis showed that observed variability was mostly between studies rather than within studies. Factors reportedly associated with higher blood EOS counts included current smoking, positive skin-prick test, elevated total IgE, comorbid allergic rhinitis, age ≤18 years, male sex, spirometric asthma/COPD diagnosis, metabolic syndrome and adiposity.ConclusionEOS distribution and range varied by study population, and were affected by clinical factors including age, smoking history and comorbidities, which, regardless of severity, should be considered during treatment decision-making.

Immunodeficiency Caused Byheterozygous RAG1 Gene Mutation in a Boy Manifested As Severe Granulomatous Diseases and Eosinophilia: A Case Report

Severe combined immunodeficiency (SCID) is an inherited diseases caused by the defective development of T- and/or B-lymphocytes refractory to common therapy. We report a 13-year old boy with combined immunodeficiency caused by a heterozygous mutation at cDNA position 1435 (c.1435C>T) in RAG1 gene. His father and grand-father carried the same heterozygous RAG1 gene mutation, and experienced anaphylactic rhinitis. His mother had mutations in 6 immunodeficiency-related heterozygous genes but no RAG1 mutation, and experienced thrombocytopenia. Both his parents had normal NK and T cell numbers, and serum immunoglobulin (Ig) levels. After birth, this patient had severe allergies to many food and drugs, and suffered from recurrent upper and lower respiratory tract infections every year. After admission, the patient complained of a generalized body rash with itches lasting about several years. We found severe granuloma lesions in the face, mouth and nose, lymphadenectasis and splenomegaly by physical examination (Figure 1a). Laboratory examinations demonstrated that he had a high eosinophilic granulocyte count as 2.19×109/L, but decreased NK cell numbers and CD4+/CD8+ T cell ratios, and IgG, IgM and IgA serum levels. This patient has been previously treated with immune-modulatory drugs including anti-histamine drugs, prednisone, γ-globulin, and chemotherapy including Cyclophosphamide, but lacking clinical improvement. In our center, after Methotrexate (1g/m2 q.w.)+L-asparaginase (10000 unit q.d.×10 days) + Dexamethasone (10mg/m2q.d.×5 days) chemotherapy, and immune enhancement therapy with γ-globulin(400mg/kg×5 days per month), the patient got his symptoms ameliorated (Figure 1b). Haploidentical hematopoietic stem cell transplantation (HCT) from his HLA 7/10 matched mother was subsequently applied, and the patient remained remission without HCT-related complications for at least 8 months after transplantation (Figure 1c). Our results thus showed the immune disorder in a boy caused by a novel heterozygous RAG1 gene c.1435C>T mutation which has been efficiently treated by our specific chemotherapy combined with HCT. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Adjuvant treatment of penetrating moxibustion at governor vessel for persistent allergic rhinitis of deficiency-cold syndrome

To explore the efficacy and action mechanism of penetrating moxibustion at governor vessel for persistent allergic rhinitis of deficiency-cold syndrome. Ninety patients with persistent allergic rhinitis of deficiency-cold syndrome were randomly divided into an observation group (n = 45) and a control group (n = 45). The patients in the control group were treated with momethasone furoate nasal spray, 2 sprays per side per time, once a day. On the basis of treatment in the control group, the patients in the observation group were additionally treated with penetrating moxibustion at governor vessel, 2 h per treatment, once a week. Both groups were treated for 4 weeks. The TCM symptom score, visual analogue scale (VAS) score and rhinoconjunctivitis quality of life questionnaire (RQLQ) score were observed in the two groups before and after treatment. The serum level of immunoglobulin E (IgE) and complete blood count of eosinophil (EOS) were measured before and after treatment, and the clinical effects were compared. Compared before treatment, the TCM symptom scores, VAS scores, RQLQ scores, serum levels of IgE and complete blood count of EOS in the two groups were all reduced after treatment (P<0.05), and those in the observation group were lower than those in the control group (P<0.05). The total effective rate in the observation group was 95.6% (43/45), which was higher than 82.2% (37/45) in the control group (P<0.05). Based on the momethasone furoate nasal spray, the adjuvant treatment of penetrating moxibustion at governor vessel could significantly improve the clinical symptoms in patients with persistent allergic rhinitis of deficiency-cold syndrome, and its mechanism may be related to the regulation of immune disorder.

Effects of an oral hydro-ethanolic purple coneflower extract on performance, clinical health and immune parameters in calves

The objective of this randomized, placebo-controlled, double-blinded field trial was to investigate the effects of oral administration of purple coneflower (Echinacea purpurea L. (EP)) on performance, health and immune parameters in calves.Calves (n = 27) were enrolled to three groups (9 calves per group): 0.5 g EP/calf per day (ECL), 5 g EP/calf per day (ECH) or placebo. Calves were vaccinated with Bluetongue-Virus (BTV) serotype 4 vaccine to investigate EPs effects on seroconversion. Clinical and performance parameters, inter alia body weight, health and milk intake were recorded for 57 days. Blood samples were analyzed for BTV antibodies and IgG by ELISA, white and red blood cell counts by flow cytometry and mRNA abundance of various inflammatory markers in leukocytes (IL-1β, IL-8, tumor necrosis factor α (TNFα), cyclooxygenase 2 (Cox-2) and prostaglandin E synthase) was studied.The findings demonstrated no differences between groups regarding performance parameters. In all groups, calves suffered from diarrhea for a minimum of 2 days, but EP reduced the number of diarrhea days by 44% in ECL and increased the body temperature. Interestingly, ECL resulted in an increased number of respiratory disease days during the follow-up period. EP did not change blood cell and IgG counts, whereas eosinophil granulocytes were reduced in ECL. Decreased levels of hemoglobin and hematocrit were found in ECH. Prostaglandin E synthase levels in leukocytes were higher in ECL and ECH, whereas no differences were obtained for IL-1β, IL-8, TNFα and Cox-2. Due to the unexpected occurrence of BTV seropositive calves before the first vaccination, 13 calves were excluded from the evaluation on seroconversion and no statistical analyses could be performed regarding antibody production. BTV-4 antibodies were not produced in 4 placebo-calves, whereas 4 of 5 and 1 of 6 ECL- and ECH-calves produced antibodies.Further investigations are needed to draw final conclusions on mode of action and efficacy of EP in calves.

The Treatment Options and Clinical Significance of Immune Thrombocytopenia Patients with Splanchnic Vein Thrombosis as the Initial Manifestation

To investigate the causes, treatment options and outcomes of immune thrombocytopenia (ITP) patients with splanchnic venous thrombosis (SVT). The clinical diagnosis, treatment and outcomes data of one 26-year-old male ITP patient with SVT as initial manifestation were collected. The possible causes and treatment options of the patients were discussed through literatures review. The result of blood routine tests of the patient showed that Plt（17-38）×109/L and eosinophils (EOS) 46.9%-50.0%. B-ultrasound and CT findings suggested thromboses of splenic vein, hepatic portal vein and its branches were formed, the patient was diagnosed as ITP with SVT. After active etiological treatment and reduced dose of low molecular weight heparin(LMWH) for anticoagulation, the patient recovered. ITP combined with large scale of SVT is rare, and it is difficult to cure. It should be pay more attention to the possible thrombosis risk triggered by a transiently increased EOS in the blood stream. Promptly etiological treatment and the balance between anticoagulant therapy and bleeding risks should be taken in clinical practice.

The effect of one bout submaximal endurance exercise on the innate and adaptive immune responses of hypertensive patients.

Changes in some of the immune system factors are associated with a risk of cardiovascular disease. Therefore, the purpose of this study was to investigate the effect of one-bout submaximal endurance exercise (OBSEE) on the innate and adaptive responses of hypertensive patients. From among 70 men with hypertension, 20 men (mean age: 60.25±4.59 years, Body Mass Index [BMI]: 29.57±3.68 kg/m2, and the maximal oxygen uptake [VO<inf>2max</inf>] 34.83±2.11 mL/kg/min) were chosen. The statistical sample performed OBSEE for 30 minutes and at 60-65% of the maximum heart rate reserve (MHRR). Blood sampling was performed to measure the response of immunoglobulin A (IgA), immunoglobulin G (IgG), immunoglobulin M (IgM), white blood cells (WBCs), neutrophil (NEUT), basophil (BASO), eosinophil (EO), monocyte (MONO) and lymphocyte (LYMPH) of hypertensive patients before and immediately after exercise. Plasma volume changes (PVCs) were also calculated in order to find out accurate effectiveness of exercise. Shapiro-Wilk test was used to normalize the research data. To analyze the data, paired sample t-test was used at significant level (P≤0.05). The hypothesis test was performed using SPSS software v. 19. The results after adjusting for PVCs showed that OBSEE significantly increased WBCs (P=0.001) in hypertensive patients. On the other hand, a significant decrease was observed in EO (P=0.001) and MONO (P=0.001) levels after OBSEE. Significant changes were not found in NEUT (P=0.072), BASO (P=0.106), LYMPH (P=0.440), IgA (P=0.382), IgG (P=0.245) and IgM (P=0.081) levels. It seems that OBSEE can reduce the risk of hypertension caused by elevated EO and MONO in hypertensive patients.

Defining type 2 asthma and patients eligible for dupilumab in Italy: a biomarker-based analysis

BackgroundAsthma is a chronic disease characterized by airway hyperresponsiveness, inflammation and mucus production. In Type 2 asthma, two phenotypic components are often co-expressed (eosinophilic and allergic). Elevated biomarker levels, such as eosinophils (EOS), fraction of exhaled nitric oxide (FeNO) and immunoglobulin E (IgE), are key clinical indicators of Type 2 inflammation. Dupilumab has been recently approved for the treatment of uncontrolled severe Type 2 asthma. Type 2 asthma includes allergic and/or eosinophilic phenotypes. The aim of this analysis was to estimate the dupilumab-eligible population in Italy and characterize it by expected biomarker status.MethodsA 4-step approach was carried out to calculate dupilumab-eligible population. The approach consisted in: (1) estimating the total number of asthma patients in Italy (using 2016–2017 Italian-adapted Global Initiative for Asthma -GINA- guidelines); (2) estimating the number of severe asthma patients with poorly controlled or uncontrolled disease (using the findings of two recent administrative claim analyses conducted in Italy); (3) stratifying the severe uncontrolled population by biomarker levels (EOS, FeNO and IgE) according to the outcomes of the QUEST trial (a clinical study assessing the efficacy of dupilumab in patients with uncontrolled moderate-to-severe asthma; NCT02414854); (4) identifying the sub-populations of severe uncontrolled asthma patients characterised by raised blood EOS and/or FeNO level (thus indicated to receive dupilumab).ResultsAccording to these estimates, about 3.3 million asthmatic patients live in Italy (6.10% of the population). Of them, almost 20 thousand (N = 19,960) have uncontrolled severe asthma. Dupilumab-eligible patients would be N = 15,988, corresponding to 80.1% of the total uncontrolled severe population. Most of these patients (89.3%; N = 14,271) have at least an increase of EOS level, while slightly more than half (51.9%; N = 8,303) have raised levels of both biomarkers. Increased FeNO levels without increased EOS are observed less frequently (N = 1,717; 10.7% of the eligible population).ConclusionsThere is a strong rationale for testing all asthma biomarkers during diagnosis and disease follow-up. Given the large availability and the limited costs, these tests are cost-effective tools to detect severe Type 2 asthma, stratify patients by phenotype, and drive appropriate treatment decisions.

The twilight zone: plasticity and mixed ontogeny of neutrophil and eosinophil granulocyte subsets.

It is now becoming clear that neutrophils and eosinophils are heterogeneous cells with potentially multiple subsets in health and disease. With greater marker coverage by multi-color flow cytometry and single-cell level sequencing of granulocyte populations, novel phenotypes of these cells began to emerge. Intriguingly, many newly described subsets blend distinctions between classical myeloid lineage phenotypes, which are especially true for tissue resident or recruited cells in contexts of inflammation and disease. This includes reports of neutrophils with features of eosinophils, monocytes and dendritic cells, and eosinophil subsets expressing neutrophil markers. Moreover, novel studies show the ability of immature neutrophils to transdifferentiate into mature cells belonging to other myeloid lineages (eosinophils, monocytes/macrophages). In this review, we summarize novel findings in this exciting research frontier and shed light on potential processes driving the plasticity and heterogeneity of granulocyte subsets. Specifically, we discuss the hematopoietic flexibility of granulocyte precursors in bone marrow and the adaptation of myeloid cells to local tissue microenvironments. The understanding of such intermediate and developmental phenotypes is very important, as it can teach us about origins of functionally distinct myeloid cells during inflammation, and explain reasons for successes and failures of biologics targeting terminally differentiated granulocytes.

Salbutamol combined with budesonide in treatment of pediatric bronchial asthma and its effect on eosinophils

BACKGROUND The aim of this study was to investigate and discuss the salbutamol combined with budesonide in treatment of pediatric bronchial asthma (BA) and its effect on eosinophils (EOS). METHODS Ninety-eight BA children admitted and treated in our hospital from July 2016 to June 2017 were collected and divided into control group (N.=49) and observation group (N.=49) according to random number table. The children in control group were treated with budesonide and those in observation group were treated with salbutamol combined with budesonide. The clinical efficacy, pulmonary functions and levels of T-lymphocyte subsets (including cluster of differentiation 3 (CD3)+, CD4+, CD8+ and CD4+/CD8+) in the immune system between two groups were compared after the treatment; the levels of eosinophil cationic protein (ECP) and eotaxin in the children were compared before the treatment and at 1, 4 and 8 weeks after the treatment; the changes in EOS counts in blood and induced sputum of the children before and after the treatment were compared, and the EOS apoptosis rate was compared at 1, 4 and 8 weeks after the treatment. RESULTS The effective rate of treatment in observation group was significantly higher than that in control group (P<0.05). After the treatment, the indexes of pulmonary function in observation group were obviously better than those in control group (P<0.05). Compared with those in control group, the levels of CD3+, CD4+ and CD4+/CD8+ of the children in observation group were elevated remarkably, while the CD8+ level was lowered (P<0.05). The levels of ECP and eotaxin in the two groups were decreased after the treatment compared with those before the treatment, and the levels in observation group were superior to those in control group (P<0.05). After the treatment, the EOS counts of both groups of children were lower than those before the treatment, and the decrease in observation group was more notable than that in control group. At 1, 4 and 8 weeks after the treatment, the EOS apoptosis rate in observation group was obviously higher than that in control group (P<0.05). CONCLUSIONS The treatment of salbutamol combined with budesonide for pediatric BA has significant therapeutic effects; it can restore the pulmonary functions rapidly and improve the immunity of the lung, reduce the levels of eotaxin, ECP and EOS of the child patients and promote EOS apoptosis.

Complex effects of whole body cryostimulation on hematological markers in patients with obesity.

BackgroundAdaptation, including changes in blood properties, to whole-body cryostimulation may depend on many factors, including body mass.AimThis study investigates whether hematological parameters change similarly in a group of people with obesity and a group of men with normal body weight after 10 and 20 cryostimulation treatments.MethodsIn our non-randomized trial, the participants were divided into two groups based on their body fat percentage: 14 men with a high (HBF = 29.35%) and 10 with a normal percent of body fat (NBF = 11.40%) and subjected to 20 whole body cryostimulation treatments (-120°C for 2–3 minutes). Blood samples were taken before the first and after the 10th and 20th cryostimulation. The following parameters were determined: red blood cells (RBC), hemoglobin concentration (HGB), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), platelets (PLT), red blood cell distribution width (RDW-SD), mean platelet volume (MPV), white blood cells (WBC), neutrophils (NEUT), lymphocytes (LYMPH), monocytes (MONO), eosinophils (EO) and basophiles (BASO).ResultsStatistically significant differences were found in red blood cells parameters such as RBC, HCT, MCV and MCHC. Time influence was noted for HCT, MCV and MCHC. Two-way ANOVA showed a significant correlation (for time and group) for 2 paramateres: RBC and MCV. For platelet parameters statistically significant differences were found for PLT (group influence) and MPV (time and group interaction). In white blood cells parameters statistically significant differences in levels of LYMPH were noted. Higher levels were observed for HBF group.ConclusionsAll observed changes were within the reference range, but hematological markers changed unevenly in people who are obese and non-obese. Therefore, it appears that an amount of fat tissue could be a factor causing the differences in adaptation to low temperature. It is suggested that 20 whole body cryostimulation sessions restore the state of homeostasis disturbed after 10 sessions.Trial registrationACTRN 12619000524190.