scholarly journals Clinical Significance of IL-23 Regulating IL-17A and/or IL-17F Positive Th17 Cells in Chronic Periodontitis

2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Zhenhua Luo ◽  
Hui Wang ◽  
Yunlong Wu ◽  
Zheng Sun ◽  
Yafei Wu
Keyword(s):  
Flow Cytometry ◽  
Correlation Coefficient ◽  
Clinical Significance ◽  
Chronic Periodontitis ◽  
Th17 Cells ◽  
Expression Level ◽  
Healthy Controls ◽  
Probing Depth ◽  
Spss Software

Objective. To investigate the expression level and clinical significance of (IL-17A+and/or IL-17F+) Th17 cells under IL-23 regulation in patients of chronic periodontitis (CP) and healthy controls (HC).Materials and Methods. The whole peripheral blood samples were collected from 30 CP patients and 25 healthy controls. Flow cytometry was used to test the (IL-17A+and/or IL-17F+) Th17 expression level. Recombinant human IL-23 (rhIL-23) was used to detect Th17 differentiation and expansion. Correlation coefficient analysis between Th17 expression level and clinical parameters was analyzed by SPSS software.Results. Flow cytometry results showed that IL-17A+IL-17F−and IL-17A−IL-17F+Th17 were both increased in CP group than in HC group (P< 0.01), while, under recombinant human IL-23 (rhIL-23) stimulation, the number of IL-17A+IL-17F−Th17 cells was significantly increased in both CP and HC groups (P< 0.01). Interestingly, IL-17A−IL-17F+Th17 cells were only increased in CP group after rhIL-23 stimulation. Additionally, correlation coefficient analysis showed significant correlation between IL-17A+IL-17F−Th17 cell and attachment loss or probing depth (P< 0.05).Conclusions. This study indicates that both the IL-17A+IL-17F−and IL-17A−IL-17F+Th17 cells may be involved in pathogenesis of periodontitis. The role of these Th17 cells in the disease pathogenesis needs to be further investigated.

Role of erythrocytes and platelets in the hypercoagulable status in polycythemia vera through phosphatidylserine exposure and microparticle generation

2013 ◽  
Vol 109 (06) ◽  
pp. 1025-1032 ◽  
Author(s):  
Chunyan Gao ◽  
Xue Yang ◽  
Jianan Li ◽  
Wei Wang ◽  
Jinxiao Hou ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Blood Cells ◽  
Significant Inhibition ◽  
Polycythaemia Vera ◽  
Factor V ◽  
Healthy Controls ◽  
Clotting Time ◽  
Thrombotic Risk ◽  
Few Data

SummaryThe development of thrombosis in polycythaemia vera (PV) involves multifactorial processes including pathological activation of blood cells. Release of microparticles (MPs) by activated cells in diseases is associated with thrombotic risk, but relatively few data are available in PV. The aim of the present study was to investigate the increase in MP release and exposure of phosphatidylserine (PS) on the outer membrane of MP-origin cells in patients with PV, and to analyse their procoagulant activity (PCA). PS-positive MPs and cells were detected by flow cytometry, while PCA was assessed with clotting time and purified coagulation complex assays. We found that PV patients had elevated circulating lactadherin+ MPs, which mostly originating from erythrocytes, platelets, granulocytes, and endothelial cells, as well as increased PS exposing erythrocytes/platelets as compared to secondary polycythaemia patients or healthy controls. These PS-bearing MPs and cells were highly procoagulant. Moreover, lactadherin competed factor V and VIII to PS and inhibited about 90% of the detected PCA in a dose-response manner while anti-TF antibody did no significant inhibition. Treatment with hydroxyurea is associated with a decrease in PS exposure and lactadherin+ MP release of erythrocytes/platelets. Our data demonstrate that PV patients are characterised by increased circulating procoagulant MPs and PS exposing erythrocytes/platelets, which could contribute to the hypercoagulable state in these patients.

scholarly journals Arginine and glutamate levels in the gingival crevicular fluid from patients with chronic periodontitis

2008 ◽  
Vol 19 (4) ◽  
pp. 318-322 ◽  
Author(s):  
Narda Téllez ◽  
Natalia Aguilera ◽  
Belkis Quiñónez ◽  
Elizabeth Silva ◽  
Luis Enrique González ◽  
...  
Keyword(s):  
Chronic Periodontitis ◽  
Gingival Crevicular Fluid ◽  
Healthy Controls ◽  
Clinical Attachment Loss ◽  
Measure Concentration ◽  
Periodontal Tissues ◽  
Glutamate Concentration ◽  
Probing Depth ◽  
Crevicular Fluid ◽  
Post Hoc

The objectives of this study were to determine arginine and glutamate levels in the gingival crevicular fluid (GCF) of adult chronic periodontitis patients versus periodontally healthy controls, and to compare two kinds of microdialysis probes: normal and U-shaped probes. The analysis of GCF components was developed to improve the diagnosis of periodontal disease (PD). Proteolysis in the periodontal tissues increases the concentration of amino acids (aa) in the GCF and the levels of these aa may reveal PD features and stages. GCF samples were collected by microdialysis in situ from 5 periodontally affected sites (probing depth >5 mm, clinical attachment loss >3 mm) in 14 adult chronic periodontitis patients and from 14 adult periodontally healthy controls. Capillary zone electrophoresis coupled to laser induced fluorescence detection was used to measure concentration of arginine and glutamate in the GCF. Data were analyzed statistically by ANOVA and Tukey's post-hoc tests (?=0.05). Arginine concentration was increased (p<0.001) and glutamate concentration was decreased (p<0.001) in chronic periodontitis patients as compared to controls. There were no significant differences (p=0.069) between the normal and U-shaped probes. In conclusion, the increase of arginine and decrease of glutamate concentration in GCF were associated to the presence of periodontitis, and might be used as markers to recognize periodontally susceptible subjects as well as to evaluate the treatment course.

scholarly journals POS0326 ROLE OF THE IL-25 / IL-17RB AXIS IN TH9 POLARIZATION IN PATIENTS WITH PROGRESSIVE SYSTEMIC SCLEROSIS

2021 ◽  
Vol 80 (Suppl 1) ◽  
pp. 390-391
Author(s):  
L. La Barbera ◽  
M. Lo Pizzo ◽  
D. DI Liberto ◽  
C. Schinocca ◽  
P. Ruscitti ◽  
...  
Keyword(s):  
Systemic Sclerosis ◽  
Th17 Cells ◽  
Lymphoid Cells ◽  
Healthy Controls ◽  
Significant Group ◽  
Th9 Cells ◽  
Inflammatory Phenotype ◽  
Over Time ◽  
Skin Samples

Background:Systemic sclerosis (SSc) is an inflammatory connective tissue disease leading to chronic and progressive fibrosis, typically affecting the skin and internal organs. The alteration of both innate and adaptive immune responses plays a pivotal role in SSc pathophysiology, although it has not yet been fully elucidated [1].Recent findings have demonstrated interleukin (IL)-9 overexpression and significant group 2 innate lymphoid cells (ILC2) expansion in patients with SSc. Th9-ILC2-mast cells axis seems to be involved in SSc tissue damage and in the induction of fibrosis [2]. Activation and production of IL-9 by Th9 cells are promoted by transforming growth factor (TGF)-β, thymic stromal lymphopoietin (TSLP), IL-25 and IL-33. Thus, the IL-25 / IL-17RB pathway would act as a key player in SSc.Objectives:The purpose of this study was to evaluate the role of the IL-25 / IL-17RB axis as a driver in Th9 polarization and ILC2 expansion and polarization in SSc patients.Methods:26 patients were enrolled in this study. Peripheral blood and skin biopsy specimens were obtained from SSc patients. PBMCs were isolated and incubated with and without recombinant (r)IL-25 for 24-48-72 hours and the frequencies of Th9 cells, Th17 cells and ILC2 were assessed by flow cytometry analysis. Moreover, the ex vivo expression of IL-17RB in ILC2 was also assessed. Immunofluorescence analysis was performed on biopsy skin samples to evaluate IL-17RB expression in ILC2.Results:In SSc samples, Th9 cells frequency progressively increased after stimulation with rIL-25, compared to healthy controls in which IL-9 frequency decreased over time regardless of rIL-25.Simultaneously, we evaluated the role of the IL-25 / IL-17RB axis in Th17 cells.In the SSc pool, the initially low rate of IL-17 increased at 72 hours after stimulation with rIL-25. In unstimulated SSc samples, the initially higher IL-17 rate decreased at 72 hours; conversely, it was consistently low in healthy controls, at both baseline and stimulated conditions.Our results confirmed the presence of IL-25-dependent clonal ILC2 expansion, suggesting a greater and progressive expansion over time in patients with SSc, compared to controls.Interestingly, increased IL-17RB expression was found in circulating ILC2 from SSc patients supporting the characterization of ILC2 inflammatory phenotype.Consistently, immunofluorescence on the skin of SSc patients showed a marked infiltrate of CD3-GATA3+ IL-17RB+ cells, confirming the presence of the activated inflammatory phenotype ILC2, absent in skin biopsies of healthy controls (Figure 1).Conclusion:These preliminary data suggest an active role of the IL-25/IL-17RB axis in SSc. It results in Th9 polarization and Th17 clonal expansion, inducing the production of IL-9 and, to a lesser extent, IL-17. Moreover, in addition to promoting Th9-mediated ILC2 differentiation, IL-25 directs the polarization of ILC2 towards the inflammatory phenotype.References:[1]Denton CP, & Khanna D. (2017). Systemic sclerosis. Lancet (London, England), 390(10103), 1685–1699.[2]Guggino G, Ciccia F, Di Liberto D, Lo Pizzo M, Ruscitti P, Cipriani P, Ferrante A, Sireci G, Dieli F, Fourniè JJ, Giacomelli R, Triolo G. s.l. Interleukin-9 over-expression and T helper 9 polarization in systemic sclerosis patients. Clin Exp Immunol, 2016 Dec.Figure 1.Immunofluorescence on biopsy skin samples of SSc patients (top) and healthy control (bottom).Disclosure of Interests:Lidia La Barbera: None declared, Marianna Lo Pizzo: None declared, Diana Di Liberto: None declared, Claudia Schinocca: None declared, Piero Ruscitti Consultant of: Pfizer, Novartis, Roche, Lilly, Celgene, Abbvie, Rorberto Giacomelli Consultant of: Pfizer, Novartis, Roche, Lilly, Celgene, Abbvie, Francesco Dieli: None declared, francesco ciccia Consultant of: Pfizer, Novartis, Roche, Lilly, Celgene, Abbvie, Giuliana Guggino Consultant of: Pfizer, Novartis, Roche, Lilly, Celgene, Abbvie

scholarly journals CircRNA ACAP2 is Overexpressed in Myocardial Infarction and Promotes the Maturation of miR-532 to Induce the Apoptosis of Cardiomyocyte

2020 ◽  
Author(s):  
Jun Zhang ◽  
Yanrong Tang ◽  
Jing Zhang ◽  
Jing Wang ◽  
Jiyun He ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Correlation Coefficient ◽  
Cell Apoptosis ◽  
Healthy Controls ◽  
Plasma Samples ◽  
Pearson’S Correlation Coefficient ◽  
Apoptosis Assay ◽  
Pearson's Correlation ◽  
Hypoxia Treatment

Abstract Background: CircRNA ACAP2 and miR-532 both promotes the apoptosis of cardiomyocytes, which contributes to myocardial infarction (MI). Therefore, ACAP2 and miR-532 may interact with each other to participate in MI. Method: Plasma samples from both MI patients (n=65) and healthy controls (n=65) were subjected to RNA extractions and RT-qPCR to analyze the expression of ACAP2, mature miR-532 and premature miR-532. Correlations among them were analyzed by Pearson’s correlation coefficient. Expression of both mature miR-532 and premature miR-532 in cardiomyocytes with ACAP2 overexpression was analyzed by RT-qPCR to study the effects of ACAP2 overexpression on the maturation of miR-532. The role of ACAP2 and miR-532 in regulating the apoptosis of cardiomyocytes induced by hypoxia was analyzed by cell apoptosis assay.Results: In this study we found that ACAP2 and mature miR-532 were both upregulated in plasma from MI patients. ACAP2 and mature miR-532 were inversely correlated, while ACAP2 and premature miR-532 were not closely correlated. In cardiomyocytes, overexpression of ACAP2 decreased the expression of mature miR-532, but not premature miR-532. Cell apoptosis analysis showed that ACAP2 and miR-532 overexpression promoted the apoptosis of cardiomyocytes induced by hypoxia treatment. In addition, miR-532 inhibitor reduced the effects of ACAP2 overexpression. Conclusion: Therefore, ACAP2 is overexpressed in MI and may promote the maturation of miR-532 to induce the apoptosis of cardiomyocyte.

scholarly journals Reactive T Cells in Convalescent COVID-19 Patients With Negative SARS-CoV-2 Antibody Serology

2021 ◽  
Vol 12 ◽  
Author(s):  
Sophie Steiner ◽  
Tatjana Schwarz ◽  
Victor M. Corman ◽  
Franziska Sotzny ◽  
Sandra Bauer ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
T Cell Response ◽  
Healthy Controls ◽  
Disease Course ◽  
Rt Pcr ◽  
Viral Control ◽  
Specific Antibodies ◽  
Mild Disease

Despite RT-PCR confirmed COVID-19, specific antibodies to SARS-CoV-2 spike are undetectable in serum in approximately 10% of convalescent patients after mild disease course. This raises the question of induction and persistence of SARS-CoV-2-reactive T cells in these convalescent individuals. Using flow cytometry, we assessed specific SARS-CoV-2 and human endemic coronaviruses (HCoV-229E, -OC43) reactive T cells after stimulation with spike and nucleocapsid peptide pools and analyzed cytokine polyfunctionality (IFNγ, TNFα, and IL-2) in seropositive and seronegative convalescent COVID-19 patients as well as in unexposed healthy controls. Stimulation with SARS-CoV-2 spike and nucleocapsid (NCAP) as well as HCoV spike peptide pools elicited a similar T cell response in seropositive and seronegative post COVID-19 patients. Significantly higher frequencies of polyfunctional cytokine nucleocapsid reactive CD4+ T cells (triple positive for IFNγ, TNFα, and IL-2) were observed in both, seropositive (p = 0.008) and seronegative (p = 0.04), COVID-19 convalescent compared to healthy controls and were detectable up to day 162 post RT-PCR positivity in seronegative convalescents. Our data indicate an important role of NCAP-specific T cells for viral control.

scholarly journals Role of microRNA-25 in high glucose cultured Müller glia

2021 ◽  
Vol 14 (5) ◽  
pp. 643-648
Author(s):  
Yu Liu ◽  
◽  
Song-Tao Yuan ◽  
Qing-Huai Liu ◽  
◽  
...  
Keyword(s):  
Flow Cytometry ◽  
High Glucose ◽  
Cultured Cells ◽  
Negative Control ◽  
Expression Level ◽  
Cell Counting ◽  
Muller Glia ◽  
Müller Glia ◽  
Proliferation And Apoptosis

AIM: To investigate the role of microRNA-25 (miR-25) in proliferation and apoptosis of retinal Müller glia (MG) under high glucose condition. METHODS: The purity of the cultured cells was verified by immunocytochemistry and flow cytometry using antibodies that specifically recognized MG. The expression level of miR-25 under normal and high glucose conditions were validated by quantitative reverse transcription polymerase chain reaction (RT-qPCR). miR-25 mimics and negative control were transfected into MG and multiple functional experiments including cell counting kit-8 assay, EDU assay, and flow cytometry were conducted to explore the effects of miR-25 on the proliferation and apoptosis of high glucose cultured MG (HGMG). RESULTS: Immunocytochemistry and flow cytometry confirmed the high purity of primary cultured MG. RT-PCR results showed that the expression level of miR-25 was significantly repressed in HGMG, while over-expression of miR-25 by miR-25 mimic markedly inhibited the high glucose induced cell apoptosis and promoted the proliferation of MG. CONCLUSION: The expression level of miR-25 is significantly downregulated in HGMG and its overexpression could attenuate the high glucose damages on MG by promoting proliferation and reducing apoptosis.

scholarly journals CircRNA circFADS2 is Downregulated in Osteoporosis and Suppresses LPS-induced Apoptosis of Osteoblast by Downregulating miR-16-5p

2020 ◽  
Author(s):  
Hailong Zhou ◽  
Jianmin Jiang ◽  
Xiaohua Chen ◽  
Zhiwei Zhang
Keyword(s):  
Correlation Coefficient ◽  
Cell Apoptosis ◽  
Cell Injury ◽  
Healthy Controls ◽  
Plasma Samples ◽  
Apoptosis Assay ◽  
Primary Osteoblasts ◽  
Induced Apoptosis ◽  
Lps Treatment

Abstract Background: CircRNA circFADS2 plays protective roles in LPS-induced cell injury, which promotes osteoporosis (OS), suggesting the involvement of circFADS2 in OS. This study aimed to explore the role of circFADS2 in OS.Methods: RT-qPCRs were performed to analyze the expression of circFADS2 and miR-16-5p in plasma samples from OS patients (n=64) and healthy controls (n=64). Correlations between circFADS2 and miR-16-5p were explored by Pearson’s correlation coefficient. In primary osteoblasts, circFADS2 and miR-16-5p were overexpressed to study the interactions between them. Cell apoptosis assay was performed to study the functions of circFADS2 and miR-16-5p in the apoptosis of osteoblasts induced by LPS.Results: CircFADS2 was downregulated in OA and inversely correlated with miR-16-5p. After treatment, circFADS2 was upregulated and miR-16-5p was downregulated. In osteoblasts, LPS treatment decreased the expression of circFADS2 but increased the expression of miR-16-5p. Overexpression of circFADS2 decreased the expression of miR-16-5p. Cell apoptosis analysis showed that circFADS2 overexpression reduced the apoptosis of osteoblasts under LPS-treatment, while miR-16-5p overexpression increased cell apoptosis. Moreover, overexpression of miR-16-5p reduced the effects of circFADS2 overexpression on the apoptosis of osteoblasts. Conclusion: Therefore, CircFADS2 is downregulated in OS and promotes LPS-induced apoptosis of osteoblast by downregulating miR-16-5p.

scholarly journals P0419AUTOANTIGEN-SPECIFIC TH17 AND TH22 INFLAME THE KIDNEY IN ANCA-VASCULITIS

2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Erika Boschmann ◽  
Tanja Hinkeldein ◽  
Andreas Kribben ◽  
Sebastian Dolff ◽  
Pieter Van Paassen ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
T Cell ◽  
Th17 Cells ◽  
Anca Vasculitis ◽  
Th22 Cells ◽  
Mrna Transcripts ◽  
Disease Induction ◽  
Renal Vasculitis

Abstract Background and Aims Anti-neutrophil-cytoplasmic-antibody (ANCA)-associated-vasculitis (AAV) is an autoimmune small-vessel-vasculitis. T-cells play a pivotal role in pathogenesis as drivers of autoantibody formation and vasculitic damage. However, the involvement of T-cells in renal vasculitis is understood poorly. IL-17C is secreted by epithelial parenchymal cells enhancing the pathogenicity of Th17 cells. Furthermore, IL-17C is a chemotactic factor for Th17 cells. The aim of this study was to assess the role of Th17 cells and IL-17C in the pathogenesis of ANCA-vasculitis in an animal model of AAV. Method Wistar-Kyoto-rats were immunized with myeloperoxidase (MPO) in Freunds Adjuvant to induce AAV. Control rats were immunized with Freunds Adjuvant only. Albuminuria was determined weekly and rats were culled after two, four and six weeks. At the time of harvest, renal T-cells were isolated and characterized by flow cytometry. Antigen-specificity was determined by ELiSPOT. Petechial bleedings on the lung surface reflecting pulmonary vasculitis were quantified after harvest. Gene expression in spleen, kidneys and lungs was studied by PCR and is expressed as fold change over controls. IL-17C levels were measured in sera by ELISA. Results MPO-rats developed detectable titres of anti-MPO by week two. By week six, all MPO-animals developed renal vasculitis with significant albuminuria and pulmonary vasculitis. Accordingly, MPO animals showed significant crescent formation as compared to the controls (% of affected glomeruli: 11.4 ±10.5% vs. 0.4 ±0.7%, p&lt;0.005). The petechial bleeding score on the lung surface was higher in MPO-immunized rats than in controls at week six (68 ±13 vs. 2 ±1, p&lt;0.05). From week two on, Th17 and Th22 cells inflamed the kidney as determined by PCR and/or flow cytometry in MPO-rats. The Th17 and Th22 infiltrate was heaviest at week six post-immunization. The intra-renal T-cell response was skewed towards Th17 as compared to the frequency of splenic Th17 cells in MPO-rats (9.1 ±4.3% vs. 1.9 ±0.6%, p&lt;0.005). The majority of intra-renal Th17 and Th22 cells was MPO-specific. Control rats did not show renal T-cell infiltration. Renal transcripts for IL-17C were slightly decreased in week 2 (0.88 ±0.1 fold), unchanged in week four (1.0 ±0.2) and slightly increased in week six after immunization (1.7 ±0.7 fold). Pulmonary IL-17C mRNA transcripts were decreased during weeks two and four after disease induction as compared to controls (0.29 ±0.07 fold, 0.98 ±0.2 fold). During week six, pulmonary IL-17C mRNA transcripts were slightly increased over controls (1.9 ±0.4 fold). Serum levels of IL17C were unchanged during weeks two to six after disease induction comparing MPO-immunized rats and controls. Conclusion Th17 and Th22 cells are drivers of renal inflammation in ANCA-vasculitis. The role of IL-17C in this cascade needs to be determined.

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