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HLA-DR and -DQ Serotyping
Published in M. Kam, Jeffrey L. Bidwell, Handbook of HLA TYPING TECHNIQUES, 2020
Principle of the Method. A lymphocyte suspension is reacted with FITC-labeled anti-immunoglobulin. This surface labels the B cells, which can be visualized using an epifluorescence microscope.23
Apoptosis: Cellular Signaling and Molecular Mechanisms
Published in John J. Lemasters, Constance Oliver, Cell Biology of Trauma, 2020
Rosemary B. Evans, John A. Cidlowski
Apoptosis is critical during B cell selection. These cells may die if heavy and light chains rearrange incorrectly (in which case no immunoglobulin is produced); they react with self antigen; they never encounter antigen; or they rearrange incorrectly during terminal differentiation. B cells are produced in the bursa of fabricius in birds, and disruption of follicular architecture or γ-irradiation of this organ causes apoptosis of these cells.78 Preneoplastic bursal stem cells expressing v-myc are hypersensitive to apoptosis induced by dispersion or radiation.78 In addition, germinal center B cells in culture undergo spontaneous apoptosis which can be reduced but not eliminated by co-culture with phorbol esters.79 This apoptosis is also inhibited by treatment of these B cells with anti-immunoglobulin antiserum. Antigen-driven selection of cells which produce antigen-specific immunoglobulin may occur in a manner similar to that in which binding of antigen to cells inhibits apoptosis. In contrast, however, some Burkitt’s lymphoma cells will undergo apoptosis if treated similarly. Anti-immunoglobulin mobilizes Ca2+ in these cells, but in the presence of EGTA apoptosis induced by Ca2+ ionophore or antisera against immunoglobulin is partially inhibited. Activation of protein kinase C with phorbol esters decreases this Ca2+-driven apoptosis to the level observed in control cultures. In addition, apoptosis in these lymphoma cells is triggered by inhibitors of protein kinase C.
Long-Acting Inhaled Beta2 Receptor Agonist Drugs
Published in Richard Beasley, Neil E. Pearce, The Role of Beta Receptor Agonist Therapy in Asthma Mortality, 2020
Studies of allergic responses in the skin indicate potential anti-inflammatory effects of the new beta2 agonists. Inhibition of early- and late-phase reactions after cutaneous injection of anti-immunoglobulin E by formoterol or terbutaline was evaluated in healthy volunteers.30,31 Both formoterol and terbutaline have an initial inhibitory effect on the flare-and-wheal reaction with a considerably greater duration of effect for formoterol (>24 h) than for terbutaline (8 h).29 The edematous late-phase reaction was also more intensively inhibited by formoterol than by terbutaline at doses that had the same inhibitory effect on the acute flare-and-wheal reactions.30 The prolonged stimulation achieved by a long-acting beta2 agonist may be more effective inhibiting inflammatory events in the tissue. Formoterol was also much more potent than salbutamol in inhibiting airway microvascular leakage induced by histamine in the guinea pig.32
Biomarkers guiding biological therapeutic strategies in chronic rhinosinusitis with nasal polyps: an emerging challenge
Published in Expert Review of Clinical Immunology, 2023
Chronic rhinosinusitis with nasal polyps (CRSwNP) is a heterogeneous inflammatory disease that is generally classified into type 2 and non-type 2 endotypes [1]. Type 2 CRSwNP is the predominant endotype of refractory CRSwNP [1]. Type 2-related cytokines (interleukin [IL]-4, IL-5, IL-9, and IL-13), mainly secreted by type 2 helper lymphocytes and group 2 innate lymphoid cells, and alarmins (IL-25, IL-33, and thymic stromal lymphopoietin [TSLP]) released from epithelial cells in response to external insults, orchestrate the complex network of type 2 inflammatory response [2,3]. With great heterogeneity and complexity in pathophysiological mechanisms, the conventional ‘one-size-fits-all’ treatment strategy does not necessarily provide benefits to all patients. To fulfill these unmet medical needs, biologics targeting type 2 inflammation have emerged to provide patients with customized therapy approaches. Currently, dupilumab (anti-IL-4 Rα monoclonal antibody [mAb]), omalizumab (anti-immunoglobulin E [IgE] mAb), and mepolizumab (anti-IL-5 mAb) are approved by the United States Food and Drug Administration and European Medicines Agency for the treatment of inadequately controlled CRSwNP. Phase III trials have been conducted or are ongoing in benralizumab (anti-IL-5 R mAb), fevipiprant (anti-prostaglandin D2 receptor 2 mAb), and tezepelumab (anti-TSLP mAb).
The intestinal quorum sensing 3-oxo-C12:2 Acyl homoserine lactone limits cytokine-induced tight junction disruption
Published in Tissue Barriers, 2020
Doriane Aguanno, Garance Coquant, Barbara G. Postal, Céline Osinski, Margaux Wieckowski, Daniel Stockholm, Jean-Pierre Grill, Véronique Carrière, Philippe Seksik, Sophie Thenet
Caco-2/TC7 cells grown on 24 mm Transwell® were fixed and permeabilized with methanol (5 min, –20°C). The following primary antibodies were used: anti-E-cadherin (rat monoclonal antibody ECCD2 M108; Takara Bio Europe; 1:1000), anti-occludin (rabbit polyclonal antibody, #71-1500; Thermofisher Scientific; 1:200), anti-tricellulin (rabbit monoclonal antibody, #700191; Thermofisher Scientific; 1:200) and anti-ZO-1 (mouse monoclonal clone 1A12, #33-9100; Thermofisher Scientific; 1:200). Alexa 488-, Alexa 546- and Alexa 647-conjugated anti-immunoglobulin G were used as secondary antibodies (1:400; Thermofisher Scientific). Nuclei were stained by 4′-6-diamidino-2-phenylindole (DAPI), and cells were examined by structured illumination microscopy using an Axio Imager 2 microscope equipped with Apotome.2, allowing optical sectioning (Zeiss, Oberkochen, Germany). Images were acquired by ZEN 2011® software (Zeiss). Quantification of fluorescent labeling was performed by measuring the staining area on thresholded images using Fiji software (https://imagej.net/Fiji/Downloads).
Effect of inhaled corticosteroid use on weight (BMI) in pediatric patients with moderate-severe asthma
Published in Journal of Asthma, 2019
Jennifer Han, John Nguyen, Yuna Kim, Bob Geng, Gale Romanowski, Lawrence Alejandro, James Proudfoot, Ronghui Xu, Sydney Leibel
In recent years, biologics, which encompass genetically designed and expressed proteins that target asthma-specific endogenous immunological processes, have become available for pediatric patients with asthma. Two biologics currently recommended for consideration in pediatric severe asthma are omalizumab and mepolizumab (16). In a randomized trial of 400 pediatric participants, omalizumab (anti-Immunoglobulin E) decreased the number of exacerbations by almost 20% and reduced the amount of inhaled glucocorticosteroid dose by 109 mcg/day compared to placebo (17). Further, in a recent “real world” observational study omalizumab was shown to decrease exacerbation frequency in patients with moderate-severe asthma with reported improved quality of life and asthma control (18). In those older than 12, mepolizumab (anti-IL5), nearly halved the amount of exacerbations in those with refractory eosinophilic asthma compared to placebo in a study with 621 participants (19). Biologic therapy may be an appropriate treatment option for patients with moderate-severe persistent asthma whose symptoms are inadequately controlled with high-dose ICS.