Upregulated Expression of Toll-Like Receptor 7 in Peripheral Blood Basophils of Patients With Allergic Rhinitis

Background Recently, it has been reported that Toll-like receptor 7 (TLR7) agonists can improve allergic rhinitis (AR) symptoms by up-regulation of Th1 cytokine release and suppression of Th2 cell functions. However, little is known of the expression of TLR7 in basophils of AR. Objective To explore the expression of TLR7 in basophils of AR, and influence of allergens on TLR7 expression. Methods The expression levels of TLR7 in basophils of patients with AR were determined by flow cytometry, and the influence of allergens on TLR7 expression was examined by real time (q) PCR. Results The percentages of TLR7+CCR3+ cells ( P < 0.001 and P = 0.011), TLR7+CD123+HLA-DR− cells ( P = 0 .016 and P = 0.042) and TLR7+CCR3+CD123+HLA-DR− cells ( P = 0.046 and P = 0.035) in blood granulocyte and mononucleated cell populations of the patients with AR were increased, respectively compared with HC subjects. TLR7 MFI on CCR3+ cells ( P = 0.050 and P = 0.043), CD123+HLA-DR− cells ( P < 0.001 and P = 0.002) and CCR3+CD123+HLA-DR− cells ( P < 0.001 and P = 0.003) were enhanced compared with HC subjects. Allergens Der p1 and OVA provoked upregulation of TLR7 expression at both protein and mRNA levels and IL-13 production in KU812 cells. House Dust Mite extract (HDME), Artemisia sieversiana wild allergen extract (ASWE), IL-31, IL-33, IL-37, and TSLP provoked elevation of IL-6 release from KU812 cells following 2 h incubation period. Conclusions The percentage of TLR7+ basophils and TLR7 expression intensity in a single basophil are both increased in the blood of patients with AR, indicating that basophils likely contribute to the pathogenesis of AR via TLR7.

Effect of flunixin or ketoprofen in caudectomy by elastration in lambs: pain and neutrophil function

ABSTRACT: Painful procedures can affect the function of innate immune cells, such as neutrophils and macrophages, increasing the risk of infectious diseases. The present work aimed to verify if the analgesics flunixin meglumine or ketoprofen can attenuate the pain/discomfort of newborn lambs submitted by elastration tail docking and thereby avoid the impairment of blood granulocytes function. Twenty-one neonate lambs were divided into three treatments: the control group (n=7), not subjected to caudectomy; the flunixin group (n=7), subjected to caudectomy under local anesthesia and analgesia with two doses of flunixin meglumine; and the ketoprofen group (n=7), subjected to caudectomy under local anesthesia and two doses of ketoprofen. Pain indicators were observed by pain posture score (PS), the number of vocalizations (V), frequency of the movement of the ears (EF), and respiratory rates (RR), observed by a 10 minutes videos for each time points: -15min, 6h, 48h, and 144h. At the same time points, the reactive oxygen species (ROS) production and phagocytosis of blood granulocytes were measured by flow cytometry. At 6h after caudectomy, there was a pain indicator increase (RR, V, and PS), a blood granulocyte percentage increase, and a granulocytes phagocytosis reduction for both groups. At 48h, the ketoprofen group spend more time in pain posture and, at 144h, they exhibited a ROS production granulocyte reduction without signs of pain. We conclude the flunixin meglumine and ketoprofen did not prevent the acute pain/discomfort caused by caudectomy, because the groups showed a pain behavior and impaired of the innate immune response however, the flunixin meglumine was effective in controlling the chronic pain and their effects on blood granulocytes function in compare ketoprofen.

Upregulated Expression of Toll Like Receptor 7 in Peripheral Blood Basophils of Patients With Allergic Rhinitis

Background: Recently, it has been reported that Toll-like receptor 7 (TLR7) agonists can improve allergic rhinitis (AR) symptoms by up-regulation of Th1 cytokine release and suppression of Th2 cell functions. However, little is known of the expression of TLR7 in basophils of AR.Objective: To explore the expression of TLR7 in basophils of AR, and influence of allergens on TLR7 expression.Methods: The expression levels of TLR7 in basophils of patients with AR were determined by flow cytometry, and the influence of allergens on TLR7 expression was examined by real time (q) PCR.Results: The percentages of TLR7+CCR3+ cells, TLR7+CD123+HLA-DR- cells and TLR7+CCR3+CD123+HLA-DR- cells in blood granulocyte and mononucleated cell populations of the patients with AR were increased, respectively compared with HC subjects. TLR7 MFI on CCR3+ cells, CD123+HLA-DR- cells and CCR3+CD123+HLA-DR- cells were enhanced. Allergens Der p1 and OVA provoked upregulation of TLR7 expression at both protein and mRNA levels and IL-13 production in KU812 cells. House Dust Mite extract (HDME), Artemisia sieversiana wild allergen extract (ASWE), IL-31, IL-33, IL-37, and TSLP provoked elevation of IL-6 release from KU812 cells following 2 h incubation period.Conclusions: The percentage of TLR7+ basophils and TLR7 expression intensity in a single basophil are both increased in the blood of patients with AR, indicating that basophils likely contribute to the pathogenesis of AR via TLR7. Trial Registration: Trial registry: Chinese clinical trial; registration number: ChiCTR-BOC-16010279.

Chemical characteristics of peripheral blood granulocyte biomembranes in frequently ill children with autonomic dysfunction syndrome

The problem of frequently ill children has extreme relevance in medical practice. A special group is FIC with autonomic dysfunction syndrome, born prematurely with perinatal damage of the central nervous system (CNS). In previous studies, we found that FIC who were born prematurely with perinatal damage of the central nervous system exhibit lower immunoreactivity than their healthy peers. Purpose of the work – to study the biochemical indicator of immunocompetent cells (the study of the phospholipid composition of granulocyte membranes) of the frequently ill children with autonomic dysfunction syndrome, born prematurely with perinatal damage to the central nervous system (CNS). Materials and methods. 68 children in the age of 6-7 years who are belonging to the group of frequently ill were examined. Out of 68 children, there were 36 children with autonomic dysfunction syndrome (SVD) born prematurely with perinatal hypoxic-ischemic damage to the central nervous system (group 1) and 32 children without autonomic dysfunction syndrome born on time without central nervous system damage (group 2). Results. A study of the phospholipid composition of blood granulocyte membranes of patients of the studied groups revealed significant differences comparison with the control group. The revealed changes in granulocyte biomembranes were more pronounced for FIC with SVD born prematurely with perinatal CNS damage than for children born on time without CNS and SVD damage. Conclusions. The obtained data allow us to conclude that FIL who were born preterm with perinatal damage of the central nervous system, changes in the biochemical characteristics of membranes are observed to a slightly greater extent than for children who were born on time without central nervous system damage. The revealed changes in immunocompetent cells are characterized by an increase in the polarity of the lipid component of biomembranes and a decrease in the relative content of neutral lipids in them. These violations lead to a change in protein-lipid interactions, which, as a result, leads to a change in the microviscosity of biomembranes and their structural and functional properties. At these children forme an inferior immune response to an infectious agent.

Blood granulocyte patterns as predictors of asthma phenotypes in adults from the EGEA study

To what extent blood granulocyte patterns may predict asthma control remains under-studied. Our aim was to study associations between blood neutrophilia and eosinophilia and asthma control outcomes in adults.Analyses were conducted in 474 asthmatics from the first follow-up of the Epidemiological Study on the Genetics and Environment of Asthma (EGEA2), including 242 asthmatics who were adults a decade earlier (EGEA1). At EGEA2, asthma control was assessed using the Global Initiative for Asthma definition (2015), and asthma exacerbations by use of urgent care or courses of oral corticosteroids in the past year. Blood EOSlo/EOShi was defined as </≥250 eosinophils·mm−3, respectively, and NEUlo/NEUhi as </≥5000 neutrophils·mm−3, respectively. Estimates were adjusted for age, sex and smoking.At EGEA2, NEUhi was associated with asthma exacerbations and poor asthma control (OR >2.10). EOShi was associated with higher bronchial hyperresponsiveness (BHR) (OR (95% CI) 2.21 (1.24–3.97)), poor lung function (p=0.02) and higher total IgE level (p=0.002). Almost 50% of asthmatics had a persistent pattern between surveys. Persistent NEUhi was associated with poor asthma control at EGEA2 (OR (95% CI) 3.09 (1.18–7.05)). EOShi at EGEA1 and persistent EOShi were associated with higher BHR (OR (95% CI) 2.36 (1.10–5.07) and 3.85 (1.11–13.34), respectively), poor lung function (p<0.06) and higher immunoglobulin E level (p<10−4) at EGEA2.Granulocyte patterns were differently associated with asthma outcomes, suggesting specific roles for each one, which could be tested as predictive signatures.

Amyloid precursor protein in peripheral granulocytes as a potential biomarker for Alzheimer’s disease

The aim of this study was to assess the potential of amyloid precursor protein in peripheral granulocytes as a diagnostic biomarker for early detection of Alzheimer’s disease. Immunohistochemistry and flow cytometry were used to evaluate amyloid precursor protein expression levels and subcellular localization in Alzheimer’s disease. Much higher amyloid precursor protein expression was observed in some leukocytes from Alzheimer’s disease patients, compared with samples from non-Alzheimer’s disease controls. In addition, flow cytometry data indicated significantly higher amyloid precursor protein expression in granulocytes from Alzheimer’s disease patients compared with control values. No statistically significant differences in amyloid precursor protein expression were obtained in lymphocytes or monocytes between the patient groups. In conclusion, amyloid precursor protein expression level in peripheral blood granulocyte is a potential biomarker for early diagnosis of Alzheimer’s disease.