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1

MacGlashan, Donald, Jane McKenzie-White, Kristine Chichester, Bruce S. Bochner, Frances M. Davis, John T. Schroeder та Lawrence M. Lichtenstein. "In Vitro Regulation of FcRIα Expression on Human Basophils by IgE Antibody". Blood 91, № 5 (1 березня 1998): 1633–43. http://dx.doi.org/10.1182/blood.v91.5.1633.

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Анотація:
Abstract In vivo studies suggested the possibility of an IgE-dependent regulation of high-affinity (FcRI) IgE receptor expression on basophils. The current studies extend these observations to in vitro cultures of human basophils. Incubation of basophils for 3 to 4 weeks resulted in a slow dissociation of IgE antibody, during which time FcRI expression decreased, as measured by flow cytometry using the anti-FcRIα monoclonal antibody, 22E7, or by measuring FcRIα mass by Western blotting of whole-cell lysates. Culture of basophils with IgE resulted in upregulation of FcRIα expression by both flow cytometry and Western blotting of whole-cell lysates. Upregulation followed a linear time course during 2 weeks of culture. The relative increase in FcRIα density depended on the starting density; with starting densities of FcRIα of 10,000 to 170,000 per basophil, the upregulation varied 20- to 1.1-fold, respectively. Upregulation occurred in high-purity basophils, was not influenced by IgG at concentrations up to 1 mg/mL, and was inhibited by dimeric IgE. Heat-inactivated IgE was less effective and the monoclonal antibody CGP51901 that prevents IgE binding to FcRIα blocked the ability of IgE to induce upregulation. The dose-response curve for IgE-induced upregulation had an effective concentration50 of 230 ng/mL. Although the receptor through which IgE induces this upregulation is not yet known, several characteristics suggest that the upregulation is mediated by IgE interacting through FcRIα itself.
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2

MacGlashan, Donald, Jane McKenzie-White, Kristine Chichester, Bruce S. Bochner, Frances M. Davis, John T. Schroeder та Lawrence M. Lichtenstein. "In Vitro Regulation of FcRIα Expression on Human Basophils by IgE Antibody". Blood 91, № 5 (1 березня 1998): 1633–43. http://dx.doi.org/10.1182/blood.v91.5.1633.1633_1633_1643.

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Анотація:
In vivo studies suggested the possibility of an IgE-dependent regulation of high-affinity (FcRI) IgE receptor expression on basophils. The current studies extend these observations to in vitro cultures of human basophils. Incubation of basophils for 3 to 4 weeks resulted in a slow dissociation of IgE antibody, during which time FcRI expression decreased, as measured by flow cytometry using the anti-FcRIα monoclonal antibody, 22E7, or by measuring FcRIα mass by Western blotting of whole-cell lysates. Culture of basophils with IgE resulted in upregulation of FcRIα expression by both flow cytometry and Western blotting of whole-cell lysates. Upregulation followed a linear time course during 2 weeks of culture. The relative increase in FcRIα density depended on the starting density; with starting densities of FcRIα of 10,000 to 170,000 per basophil, the upregulation varied 20- to 1.1-fold, respectively. Upregulation occurred in high-purity basophils, was not influenced by IgG at concentrations up to 1 mg/mL, and was inhibited by dimeric IgE. Heat-inactivated IgE was less effective and the monoclonal antibody CGP51901 that prevents IgE binding to FcRIα blocked the ability of IgE to induce upregulation. The dose-response curve for IgE-induced upregulation had an effective concentration50 of 230 ng/mL. Although the receptor through which IgE induces this upregulation is not yet known, several characteristics suggest that the upregulation is mediated by IgE interacting through FcRIα itself.
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3

HILL, M. R., M. R. NEWTON, and B. J. HART. "Comparative IgE responses to extracts of five species of house dust mite, using Western blotting." Clinical & Experimental Allergy 23, no. 2 (February 1993): 110–16. http://dx.doi.org/10.1111/j.1365-2222.1993.tb00305.x.

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4

Resende, Rafael de Oliveira, Leandro Hideki Ynoue, Juliana Silva Miranda, Karine Cristine de Almeida, Deise Aparecida de Oliveira Silva, Monica Camargo Sopelete, Ronaldo Alves, Margareth Leitão Gennari-Cardoso, and Ernesto Akio Taketomi. "IgE, IgG1, and IgG4 Reactivity to Dermatophagoides pteronyssinus Glycosylated Extract in Allergic Patients." BioMed Research International 2019 (July 29, 2019): 1–12. http://dx.doi.org/10.1155/2019/9840890.

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Анотація:
Background. House dust mites are important allergen sources and some of these allergenic proteins may contain carbohydrate moieties, which are able to be isolated using lectins, as Concanavalin A (ConA). This study aimed to investigate allergenicity (IgE) and antigenicity (IgG1 and IgG4) of ConA-unbound and ConA-bound Dermatophagoides pteronyssinus (Dpt) crude extracts using sera of mite-allergic patients as well as inhibition capacity of antibody binding. Material and Methods. We obtained mannose-enriched and mannose-depleted fractions from Dpt by ConA affinity chromatography. Both ConA-bound and ConA-unbound fractions were evaluated by ELISA and Western Blotting for specific IgE, IgG1, and IgG4 reactivity with sera obtained from 95 mite-allergic patients (DP+) and 92 nonallergic (NA) subjects. Inhibition ELISA was used to assess cross-reactivity between Dpt extract and its fractions. Results. Among the DP+ patients, no difference was found between ConA-unbound and ConA-bound fractions regarding the levels of specific IgE, IgG1, and IgG4. Nonallergic subjects had the same levels of specific IgG1 to both ConA-unbound and ConA-bound fractions, although for specific IgG4, values were higher for ConA-bound. A positive correlation was found among specific IgE, IgG1, and IgG4 levels when Dpt was compared to ConA-unbound and ConA-bound fractions. Recognition of crude Dpt by IgE, IgG1, and IgG4 was highly inhibited by ConA-unbound and ConA-bound fractions. Western Blotting revealed a broad spectrum of bands ranging from 14 to 116 kDa recognized by specific IgE and IgG4. However, IgG1 reached higher frequency values on high molecular weight polypeptides. Conclusion. ConA-unbound and ConA-bound fractions derived from D. pteronyssinus crude extract revealed important components involved in the IgE recognition in allergic patients as well as IgG1 and/or IgG4 in allergic and healthy subjects.
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5

Seminario, Maria-Cristina, Sarbjit S. Saini, Donald W. MacGlashan та Bruce S. Bochner. "Intracellular Expression and Release of FcεRIα by Human Eosinophils". Journal of Immunology 162, № 11 (1 червня 1999): 6893–900. http://dx.doi.org/10.4049/jimmunol.162.11.6893.

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Анотація:
Abstract Although FcεR have been detected on human eosinophils, levels varied from moderate to extremely low or undetectable depending on the donor and methods used. We have attempted to resolve the conflicting data by measuring levels of IgE, FcεRI, and FcεRII in or on human eosinophils from a variety of donors (n = 26) and late-phase bronchoalveolar lavage fluids (n = 5). Our results demonstrated little or no cell surface IgE or IgE receptors as analyzed by immunofluorescence and flow cytometry. Culture of eosinophils for up to 11 days in the presence or absence of IgE and/or IL-4 (conditions that enhance FcεR on other cells) failed to induce any detectable surface FcεR. However, immunoprecipitation and Western blot analysis of eosinophil lysates using mAb specific for FcεRIα showed a distinct band of approximately 50 kDa, similar to that found in basophils. Western blotting also showed the presence of FcR γ-chain, but no FcεRIβ. Surface biotinylation followed by immunoprecipitation again failed to detect surface FcεRIα, although surface FcRγ was easily detected. Since we were able to detect intracellular FcεRIα, we examined its release from eosinophils. Immunoprecipitation and Western blotting demonstrated the release of FcεRIα into the supernatant of cultured eosinophils, peaking at approximately 48 h. We conclude that eosinophils possess a sizable intracellular pool of FcεRIα that is available for release, with undetectable surface levels in a variety of subjects, including those with eosinophilia and elevated serum IgE. The biological relevance of this soluble form of FcεRIα remains to be determined.
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6

Feng, Yufei, and Lei Jiang. "Analysis of Immunogenicity of Acanthopanax senticosus Injection." Natural Product Communications 15, no. 7 (July 2020): 1934578X2093714. http://dx.doi.org/10.1177/1934578x20937144.

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Анотація:
The cause of the immunogenicity of Acanthopanax senticosus injection has been studied. The method used for the preparation of the injection includes boiling the plant material in water, precipitation with ethanol, and filtration to produce the extract. In the current study, this extract was treated with saturated ammonium sulfate to collect the protein from the extract. Indicators of systemic allergy, including total immunoglobulin (IgE), interleukin -4 (IL-4), and interferon-gamma (IFN-γ) levels in the antiserum of Guinea pigs sensitized with this protein extract were used to determine the immunogenicity of the protein extract. Testing was performed using Western blotting to identify IgE in antiserum binding in Guinea pigs sensitized by injection. The target protein band was further identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry. The protein extract from A. senticosus caused positive reactions and significantly increased the levels of total IgE, IL-4, and IFN-γ in the serum of sensitized Guinea pigs. Western blotting showed that a group of proteins with a molecular weight of 60 000 in the A. senticosus protein extract acted as the major immunogenic protein by binding to serum IgE of Guinea pigs sensitized with A. senticosus injection. The protein was a pyruvate kinase homologous protein with high sequence similarity to the pyruvate kinase protein of Vitis vinifera. It was concluded that the allergic reaction caused by the injection of A. senticosus may be related to the interaction of these macromolecular proteins within the body.
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7

Schramm, Gabriele, Helga Kahlert, Roland Suck, Bernhard Weber, Hans-Thomas Stüwe, Wolf-Dieter Müller, Albrecht Bufe, et al. "“Allergen Engineering”: Variants of the Timothy Grass Pollen Allergen Phl p 5b with Reduced IgE-Binding Capacity but Conserved T Cell Reactivity." Journal of Immunology 162, no. 4 (February 15, 1999): 2406–14. http://dx.doi.org/10.4049/jimmunol.162.4.2406.

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Анотація:
Abstract One problem of conventional allergen-specific immunotherapy is the risk of anaphylactic reactions. A new approach to make immunotherapy safer and more efficient might be the application of engineered allergens with reduced IgE-binding capacity but retained T cell reactivity. Using overlapping dodeca-peptides, the dominant T cell epitopes of the timothy grass pollen allergen Phl p 5b were identified. By site-directed mutagenesis outside these regions, point and deletion mutants were generated. Allergen variants were analyzed for IgE-binding capacity with sera of different grass pollen allergic patients by Western blotting, Dot blotting, and EAST inhibition test, and for histamine releasing capacity with peripheral blood basophils from different patients. The deletion mutants revealed significantly reduced IgE reactivity and histamine releasing capacity, compared with the wild-type Phl p 5b. Furthermore, in vivo skin prick tests showed that the deletion mutants had a significantly lower potency to induce cutaneous reactions than the wild-type Phl p 5b. On the other hand, T cell clones and T cell lines from different allergic patients showed comparable proliferation after stimulation with allergen variants and wild-type Phl p 5b. Considering their reduced anaphylactogenic potential together with their conserved T cell reactivity, the engineered allergens could be important tools for efficient and safe allergen-specific immunotherapy.
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8

Guiraldelli, Michel F., Elsa H. Berenstein, and Reuben P. Siraganian. "A Monoclonal Antibody that inhibits IgE binding to the High affinity IgE receptor recognizes a glycolipid (36.11)." Journal of Immunology 182, no. 1_Supplement (April 1, 2009): 36.11. http://dx.doi.org/10.4049/jimmunol.182.supp.36.11.

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Анотація:
Abstract The mAb BD6 is a monoclonal IgM that binds to the surface of RBL-2H3 cells and inhibits the binding of IgE to FcεRI, without reacting with this receptor. By cloning, mAb BD6 binding was resulted by the expression of the alpha1,3-galactosyltransferase gene. In binding assays on RBL-2H3 and alpha1,3-galactosyltransferase transfected PEAK cells there was partial competition between mAb BD6 and IB4, a galactose binding lectin. Both galactose and melibiose decreased the binding of mAb BD6 in a dose dependent manner on RBL-2H3 cells and abolished its binding on alpha1,3-galactosyltransferase transfected PEAK cells. MAb BD6 recognized a low molecular weight band by immunoblotting suggesting that it may be binding a lipid. This was confirmed by blotting lipids from RBL-2H3 cells separated by thin layer chromatography. By sucrose gradient analysis mAb BD6 bound to the lipid rafts fractions. RBL-2H3 variants were isolated completely deficient in mAb BD6 binding; these cells were also deficient in the ganglioside GD1b and had low expression of ganglioside GM1. However, these cells deficient in mAb BD6 binding and GD1b ganglioside still had normal FceRI induced degranulation. These results demonstrate that mAb BD6 inhibits IgE binding and reacts with a glycolipid present in lipid rafts; furthermore the ganglioside GD1b is not essential for degranulation. Supported by the Intramural Research Program, NIDCR, NIH.
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9

Peng, C., F. M. Davis, L. K. Sun, R. S. Liou, Y. W. Kim, and T. W. Chang. "A new isoform of human membrane-bound IgE." Journal of Immunology 148, no. 1 (January 1, 1992): 129–36. http://dx.doi.org/10.4049/jimmunol.148.1.129.

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Анотація:
Abstract The epsilon-chain of membrane-bound IgE on the surface of B lymphocytes is known to contain a membrane-anchoring peptide segment that is encoded by two membrane exons, me.1 and me.2. In analyzing pertinent segments in mRNA from human IgE-expressing B cells by using PCR methods and Northern blotting analyses, we have identified three species of mRNA of epsilon-chain with variations in the splicing of the membrane exons. The conventional species (m/s) contains the predicted me.1 and me.2; species m/1 harbors 156 extra nucleotides 5' of me.1 with unaltered reading frame; species s/t lacks me.1 and hence the segment encoding the hydrophobic transmembrane stretch and contains a shifted me.2 reading frame. Rabbit antibodies, which were prepared by immunization using a peptide of 36 amino acid residues representing an encoded segment unique to mRNA species m/l, could specifically bind to human IgE-expressing B cell lines and react with an epsilon-chain on Western immunoblots. These results indicate that there exists a previously unidentified isoform of human membrane-bound IgE.
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10

Plundrich, Nathalie, Mary Ann Lila, Edward Foegeding, and Scott Laster. "Protein-bound polyphenols create “ghost” band artifacts during chemiluminescence-based antigen detection." F1000Research 6 (March 13, 2017): 254. http://dx.doi.org/10.12688/f1000research.10622.1.

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Анотація:
Antigen detection during Western blotting commonly utilizes a horseradish peroxidase-coupled secondary antibody and enhanced chemiluminescent substrate. We utilized this technique to examine the impact of green tea-derived polyphenols on the binding of egg white protein-specific IgE antibodies from allergic human plasma to their cognate antigens. Our experiments unexpectedly showed that green tea-derived polyphenols, when stably complexed with egg white proteins, caused hyperactivation of horseradish peroxidase resulting in the appearance of white “ghost” bands. This study suggests that caution should be taken when evaluating polyphenol-bound proteins by enhanced chemiluminescence Western blotting using horseradish peroxidase and demonstrates that protein-bound polyphenols can be a source of “ghost” band artifacts on Western blots.
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11

Plundrich, Nathalie, Mary Ann Lila, Edward Foegeding, and Scott Laster. "Protein-bound polyphenols create “ghost” band artifacts during chemiluminescence-based antigen detection." F1000Research 6 (May 26, 2017): 254. http://dx.doi.org/10.12688/f1000research.10622.2.

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Анотація:
Antigen detection during Western blotting commonly utilizes a horseradish peroxidase-coupled secondary antibody and enhanced chemiluminescent substrate. We utilized this technique to examine the impact of green tea-derived polyphenols on the binding of egg white protein-specific IgE antibodies from allergic human plasma to their cognate antigens. Our experiments unexpectedly showed that green tea-derived polyphenols, when stably complexed with egg white proteins, caused “ghost” band formation in the presence of horseradish peroxide. This study suggests that caution should be taken when evaluating polyphenol-bound proteins by enhanced chemiluminescence Western blotting using horseradish peroxidase and demonstrates that protein-bound polyphenols can be a source of “ghost” band artifacts on Western blots.
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12

Ford, S. A., B. A. Baldo, R. Panzani, and D. Bass. "Cypress (Cupressus sempervirens) Pollen Allergens: Identification by Protein Blotting and Improved Detection of Specific IgE Antibodies." International Archives of Allergy and Immunology 95, no. 2-3 (1991): 178–83. http://dx.doi.org/10.1159/000235426.

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13

Fan, Meiqi, Nishala Erandi Wedamulla, Young-Jin Choi, Qun Zhang, Sung Mun Bae, and Eun-Kyung Kim. "Tenebrio molitor Larva Trypsin Hydrolysate Ameliorates Atopic Dermatitis in C57BL/6 Mice by Targeting the TLR-Mediated MyD88-Dependent MAPK Signaling Pathway." Nutrients 15, no. 1 (December 24, 2022): 93. http://dx.doi.org/10.3390/nu15010093.

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Анотація:
Atopic dermatitis (AD) is a widely researched chronic inflammatory skin disease with a complex etiology. The increased prevalence of AD necessitates exploration of natural sources as potential therapeutic agents with limited side effects. In the current study, a 1-chloro-2,4-dinitrobenzene (DNCB)-induced AD mouse model was used to examine the anti-AD effects of Tenebrio molitor trypsin hydrolysate (TMTH) and its underlying molecular mechanism. DNCB-treated mice were treated with TMTH (1 and 10 mg/kg), and prednisolone (3 mg/kg) was used as the positive control. Serum and skin tissue samples were collected for subsequent analyses. The expression levels of proteins linked to the myeloid differentiation primary response 88 (MyD88)-dependent mitogen-activated protein kinase (MAPK) signaling pathway and serum IgE levels were estimated via Western blotting technique and ELISA (enzyme-linked immunosorbent assay), respectively. Inflammatory cell infiltration and thickening of the dorsal skin were measured using toluidine blue and hematoxylin and eosin staining, respectively. Oral administration of TMTH significantly reduced mast cell infiltration and dermal and epidermal thickness. Moreover, TMTH treatment reduced serum IgE levels. Western blotting confirmed that TMTH treatment suppressed the MyD88-dependent MAPK signaling pathway. Therefore, TMTH substantially inhibited AD-like skin lesion formation via immunomodulation, showing considerable potential for AD treatment.
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14

Gangur, Venugopal, Rick Jorgensen, Haoran Gao, Yining Jin, Jillian Salloum, Dan Jian, and Perry KW Ng. "Characterization of protein allergens in a mouse model of wheat-induced anaphylaxis." Journal of Immunology 202, no. 1_Supplement (May 1, 2019): 55.5. http://dx.doi.org/10.4049/jimmunol.202.supp.55.5.

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Анотація:
Abstract Wheat allergies are among the major types of food allergies that are potentially life-threatening because of anaphylaxis. Wheat contains 3 different types of protein allergens: water/saline-soluble albumins/globulins, alcohol-soluble gliadins and acid-soluble glutenins. Although gliadins have been linked to anaphylaxis, whether or not other proteins cause anaphylaxis is not well understood. Here, we sought to identify saline-soluble wheat proteins (SSWP) involved in anaphylaxis using a mouse model. Balb/cJ mice were sensitized to SSWP by four intraperitoneal (IP) injections along with alum as an adjuvant. Sensitization was confirmed by testing for SSWP-specific IgE using ELISA. Anaphylaxis was measured by hypothermia shock response. After multiple booster injections with SSWP, hyper-IgE plasma was collected and pooled to create a mini-plasma bank. Using hyper-IgE plasma, an IgE-Western blotting (WB) method was optimized. The IgE-binding proteins were identified by sequencing using LC/MS/MS method. The hyper-IgE plasma had an IgE titer of ~2560. The optimized IgE-WB included 3 overnight incubations followed by signal detection using an HRPO-based method. Sequencing of the IgE binding bands in WB identified the following proteins: globulin 3A (66 kDa), globulin 3B (57 kDa), serpin (43 kDa), glyceraldehyde-3-phosphate dehydrogenase (37 kDa), chitinase (28 kDa) and globulin 1 (25 kDa). This is the first study characterizing salt-soluble protein allergens in a mouse model of wheat-induced anaphylaxis. Funding: USDA, NIFA, Hatch project MICL02486 (Accession Number: 1012322), Agricultural and Food Research Initiative Competitive Program, grant number: 2018-67017-27876.
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15

Camargo Hizume-Kunzler, Deborah, Flavia R. Greiffo, Bárbara Fortkamp, Gabriel Ribeiro Freitas, Juliana Keller Nascimento, Thayse Regina Bruggemann, Leonardo Melo Avila, et al. "Aerobic Exercise Decreases Lung Inflammation by IgE Decrement in an OVA Mice Model." International Journal of Sports Medicine 38, no. 06 (April 7, 2017): 473–80. http://dx.doi.org/10.1055/s-0042-121638.

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Анотація:
AbstractAerobic exercise (AE) reduces lung function decline and risk of exacerbations in asthmatic patients. However, the inflammatory lung response involved in exercise during the sensitization remains unclear. Therefore, we evaluated the effects of exercise for 2 weeks in an experimental model of sensitization and single ovalbumin-challenge. Mice were divided into 4 groups: mice non-sensitized and not submitted to exercise (Sedentary, n=10); mice non-sensitized and submitted to exercise (Exercise, n=10); mice sensitized and exposed to ovalbumin (OVA, n=10); and mice sensitized, submitted to exercise and exposed to OVA (OVA+Exercise, n=10). 24 h after the OVA/saline exposure, we counted inflammatory cells from bronchoalveolar fluid (BALF), lung levels of total IgE, IL-4, IL-5, IL-10 and IL-1ra, measurements of OVA-specific IgG1 and IgE, and VEGF and NOS-2 expression via western blotting. AE reduced cell counts from BALF in the OVA group (p<0.05), total IgE, IL-4 and IL-5 lung levels and OVA-specific IgE and IgG1 titers (p<0.05). There was an increase of NOS-2 expression, IL-10 and IL-1ra lung levels in the OVA groups (p<0.05). Our results showed that AE attenuated the acute lung inflammation, suggesting immunomodulatory properties on the sensitization process in the early phases of antigen presentation in asthma.
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16

Mortaz, Esmaeil, Gert Folkerts, Ferdi Engels, Frans P. Nijkamp, and Frank A. Redegeld. "Cigarette smoke suppresses in vitro allergic activation of mouse mast cells (80.1)." Journal of Immunology 182, no. 1_Supplement (April 1, 2009): 80.1. http://dx.doi.org/10.4049/jimmunol.182.supp.80.1.

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Анотація:
Abstract Abstract Background: Mast cells are important effector cells in innate or acquired immunity that contribute to host defense. Excessive activation of mast cells can result in the development of allergic diseases, including atopic asthma. Mast cell activation by IgE and specific antigen induces the cells to release spasmogenic, vasoactive, and proinflammatory mediators, which enhance airway smooth muscle contraction, vascular permeability, and inflammatory cell recruitment. Recently, we have demonstrated that exposure of mast cells to cigarette smoke medium (CSM) triggered mast cells to produce chemokines. On the other hand, smoking may decrease the risk of allergic sensitization, which could be explained by a reduced IgE production or a diminished response of mast cells to activation of the IgE receptor. Objective: In this study, we investigated the effect of CSM on allergic activation of mast cells through IgE and antigen. Methods: Primary cultured murine mast cells were exposed to CSM and activated with IgE and antigen or lipopolysaccharide (LPS). The releases of granules, production of leukotrienes, chemokines and cytokines was determined in supernatants by ELISA. The effect of CSM exposure on intracellular signaling, especially the NF-κB and Erk1/2 pathways, was analyzed by Western blotting. Results: CSM suppressed IgE-mediated degranulation and cytokine release, but no effect was observed on leukotriene release. CSM induced phosphorylation of Erk1/2 in mast cells. In CSM-exposed mast cells ATF-1 was phosphorylated after stimulation with IgE/Ag. LPS activated mast cells were not influenced by CSM. Conclusion: Our study suggests that exposure to cigarette smoke may lead to a reduced allergic activation of mast cells without affecting their response to activation via e.g. bacterial derived LPS.
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17

Chang, Tsong-Min, Tzu-Chih Hsiao, Ting-Ya Yang та Huey-Chun Huang. "IgE-Induced Mast Cell Activation Is Suppressed by Dihydromyricetin through the Inhibition of NF-κB Signaling Pathway". Molecules 26, № 13 (25 червня 2021): 3877. http://dx.doi.org/10.3390/molecules26133877.

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Анотація:
Mast cells play a crucial role in the pathogenesis of type 1 allergic reactions by binding to IgE and allergen complexes and initiating the degranulation process, releasing pro-inflammatory mediators. Recently, research has focused on finding a stable and effective anti-allergy compound to prevent or treat anaphylaxis. Dihydromyricetin (DHM) is a flavonoid compound with several pharmacological properties, including free radical scavenging, antithrombotic, anticancer, and anti-inflammatory activities. In this study, we investigated the anti-allergic inflammatory effects and the underlying molecular mechanism of DHM in the DNP-IgE-sensitized human mast cell line, KU812. The cytokine levels and mast cell degranulation assays were determined by enzyme-linked immunosorbent assay (ELISA). The possible mechanism of the DHM-mediated anti-allergic signaling pathway was analyzed by western blotting. It was found that treatment with DHM suppressed the levels of inflammatory cytokines TNF-α and IL-6 in DNP-IgE-sensitized KU812 cells. The anti-allergic inflammatory properties of DHM were mediated by inhibition of NF-κB activation. In addition, DHM suppressed the phosphorylation of signal transducer and activator of transcription 5 (STAT5) and mast cell-derived tryptase production. Our study shows that DHM could mitigate mast cell activation in allergic diseases.
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18

Zhang, K., A. Saxon, and E. E. Max. "Two unusual forms of human immunoglobulin E encoded by alternative RNA splicing of epsilon heavy chain membrane exons." Journal of Experimental Medicine 176, no. 1 (July 1, 1992): 233–43. http://dx.doi.org/10.1084/jem.176.1.233.

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Анотація:
We present evidence for RNA transcripts encoding two forms of human epsilon immunoglobulin (Ig) heavy chain that differ significantly from those of other isotypes. We previously demonstrated three human epsilon mRNA species, instead of the two, corresponding to membrane and secreted proteins, seen with other heavy chain transcripts. In human genomic DNA downstream of the C epsilon gene, we identified sequences homologous to the two putative murine exons M1 (encoding a hydrophobic, presumably transmembrane region) and M2 (encoding hydrophilic residues). To determine the structures of epsilon transcripts containing these sequences, we amplified epsilon-related RNAs with the reverse transcriptase polymerase chain reaction. RNA was examined from fresh human B cells stimulated to IgE production by interleukin 4 plus anti-CD40, as well as from the human IgE-producing line AF10. Instead of the single CH4-M1-M2 splice product predicted for murine membrane IgE, we found two other RNA species. One form has the structure CH4-M1'-M2, in which M1' includes the human sequence homologous to the murine M1 as well as a unique segment of 52 codons further upstream in the genomic sequence; this RNA species apparently encodes the IgE expressed on the membrane of IgE-producing lymphocytes. The other RNA has the structure CH4-M2', in which M2' is spliced in an alternative reading frame that includes an additional 109 codons downstream of the termination codon of the CH4-M1'-M2 form. Because the CH4-M2' mRNA form does not encode a hydrophobic segment, its translated product should be secreted. A secreted epsilon protein of approximately the size predicted for this form was identified by Western blotting. This novel IgE protein could play a significant and distinctive role in allergic disorders.
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19

Ito, Yukiko, Yoshiyuki Yoshinaka, Masuichi Ohi, and Yasuo Sakakura. "Analysis by Electrophoretic Transfer Blotting of Japanese Cedar Pollen Allergens which React with IgG and IgE Antibodies in the Serum of Patients." International Archives of Allergy and Immunology 81, no. 2 (1986): 174–79. http://dx.doi.org/10.1159/000234128.

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20

Barrera, Coralie, Bénédicte Richaud-Thiriez, Steffi Rocchi, Bénédicte Rognon, Sandrine Roussel, Frédéric Grenouillet, Audrey Laboissière, Jean-Charles Dalphin, Gabriel Reboux, and Laurence Millon. "New Commercially Available IgG Kits and Time-Resolved Fluorometric IgE Assay for Diagnosis of Allergic Bronchopulmonary Aspergillosis in Patients with Cystic Fibrosis." Clinical and Vaccine Immunology 23, no. 3 (December 23, 2015): 196–203. http://dx.doi.org/10.1128/cvi.00498-15.

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Анотація:
ABSTRACTAllergic bronchopulmonary aspergillosis (ABPA) is difficult to diagnose; diagnosis relies on clinical, radiological, pathological, and serological criteria. Our aim was to assess the performance of two new commercially available kits and a new in-house assay: anAspergillus fumigatusenzyme-linked immunosorbent assay (ELISA) IgG kit (Bordier Affinity Products), anAspergillusWestern blotting IgG kit (LDBio Diagnostics), and a new in-house time-resolved fluorometric IgE assay (dissociation-enhanced lanthanide fluorescent immunoassay, or DELFIA) using recombinant proteins from anAspergillussp. recently developed by our laboratory for ABPA diagnosis in a retrospective study that included 26 cystic fibrosis patients.Aspergillus fumigatus-specific IgG levels measured by a commercial ELISA kit were in accordance with the level of precipitins currently used in our lab. The ELISA kit could accelerate and help standardize ABPA diagnosis.Aspergillus fumigatus-specific IgE levels measured by ImmunoCAP (Phadia) withA. fumigatusM3 antigen and by DELFIA with a purified protein extract ofA. fumigatuswere significantly correlated (P< 10−6). The results with recombinant antigens glucose-6-phosphate isomerase and mannitol-1-phosphate dehydrogenase were encouraging but must be confirmed with sera from more patients. The DELFIA is an effective tool that can detect specific IgE against more fungal allergens than can be detected with other commercially available tests.
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21

Choi, Wooram, Hwa Pyoung Lee, Philaxay Manilack, Veosavanh Saysavanh, Byoung-Hee Lee, Sarah Lee, Eunji Kim, and Jae Youl Cho. "Antiallergic Effects of Callerya atropurpurea Extract In Vitro and in an In Vivo Atopic Dermatitis Model." Plants 12, no. 4 (February 14, 2023): 860. http://dx.doi.org/10.3390/plants12040860.

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Анотація:
(1) Background: Callerya atropurpurea is found in Laos, Thailand, and Vietnam. Although the anti-inflammatory action of C. atropurpurea has been investigated, the functions of this plant in allergic responses are not understood. Here, we explored the antiallergic mechanism of C. atropurpurea ethanol extract (Ca-EE) using in vitro assays and an in vivo atopic model. (2) Methods: The constituents of Ca-EE were analyzed using GC/MS. Inhibition of lipoxygenase and β-hexosaminidase activity was examined, and the expression of inflammatory genes was measured by quantitative real-time PCR. The regulatory roles of Ca-EE in IgE/FcεRI signaling were examined by Western blotting. The DNCB-induced atopic dermatitis mouse model was performed with histological analysis. (3) Results: Ca-EE comprised cis-raphasatin, lupeol, some sugars, and fatty acids. In RBL-2H3 cells, treatment with Ca-EE significantly reduced the activities of lipoxygenase and β-hexosaminidase, as well as cytokine gene expression. IgE-mediated signaling was downregulated by blocking Lyn kinases. Moreover, Ca-EE effectively inhibited allergic symptoms in the DNCB-induced atopic dermatitis model without toxicity. (4) Conclusions: Ca-EE displayed antiallergic activities through regulating IgE/Lyn signaling in RBL-2H3 cells and a contact dermatitis model. These results indicate that Ca-EE could be effective for allergic disease treatment.
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22

Ro, Jai, Yun Song Lee, Soo Youl Kim, and Gwan Hong. "Transglutaminase 2 expressed in mast cell activation induces IgE production in B cells, airway inflammation and remodeling via up-regulating CD40L and cytokine expression in mouse allergic asthma (64.4)." Journal of Immunology 188, no. 1_Supplement (May 1, 2012): 64.4. http://dx.doi.org/10.4049/jimmunol.188.supp.64.4.

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Abstract TGase 2 is over-expressed in a variety of inflammatory diseases including allergic asthma. This study aimed to investigate the role of TGase 2 in signaling pathways in mast cells related to OVA-induced allergic asthma in vitro and in vivo. BMMCs isolated from C57BL/6 WT or TGase 2-/- mice were activated with Ag/Ab. Mice were sensitized and challenged with OVA to induce asthma. [Ca2+]i level were determined by confocal, mast cells by May-Giemsa, LT, IgE and cytokine levels by ELISA, expression of TGase 2, PLA2, EMBP, Muc5ac, collagen, cytokines, MMPs and TIMPs by Western blotting or RT-PCR, NF-kB activity by EMSA, the recruitment of inflammatory cells into BAL fluid or lung tissues by Diff-Quik and H&E, AHR by Fexivent system. Act-WT-BMMCs expressed TGase 2, and increased the [Ca2+]i level, activities of I-kB, NF-kB and PLA2, LTs level, expression of CD40/CD40L, cytokines and MMPs, collagen deposition, and IgE production in act-B cells, whereas act-KO-BMMCs reversed all of them. TGase 2-/- mice protected against OVA-specific IgE production as well as AHR, expression of signaling molecules and mediator release in BAL cells and lung tissues versus those in WT mice. Our data suggest that TGase 2 expression and Ca2+ influx required by bidirectional events in mast cell activation facilitate IgE production in B cells via regulating mast cell CD40L expression, and induce the expression of numerous molecules associated with airway inflammation and remodeling in allergic asthma.
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23

Sasso, E. H., G. J. Silverman, and M. Mannik. "Human IgM molecules that bind staphylococcal protein A contain VHIII H chains." Journal of Immunology 142, no. 8 (April 15, 1989): 2778–83. http://dx.doi.org/10.4049/jimmunol.142.8.2778.

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Abstract Staphylococcal protein A (SPA) is a bacterial membrane protein which has distinct binding sites for Fc gamma and for the Fab region of some IgM, IgG, IgA, and IgE molecules. This study establishes a structure-function correlation responsible for the binding of Ig Fab regions to SPA. Binding of 24 isolated human monoclonal IgM proteins to SPA was measured in a solid phase RIA. VH and V kappa subgroups of each IgM were determined by SDS-PAGE, transfer blotting, and detection with antisera prepared against specific first framework region peptides. Binding to SPA was seen with 10 of 11 VHIII IgM, but none of the 7 VHI or 6 VHII. Similarly, polyclonal IgM fractionated on a SPA-Sepharose CL4B column showed nearly complete partition of VHIII molecules into the SPA-binding fraction, and VHI and VHII subgroup proteins into the fall-through. We conclude that SPA binding is a functional marker for VHIII H chains in human IgM molecules.
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24

Lin, Tong-Jun, Yong Jun Yang, Jeffery D. Molkentin, and Jason N. Berman. "Egr1-Rcan1 axis serves as a molecular switch from activation to inhibition in IgE-mediated signalling (139.3)." Journal of Immunology 182, no. 1_Supplement (April 1, 2009): 139.3. http://dx.doi.org/10.4049/jimmunol.182.supp.139.3.

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Abstract Mast cells play a central role in allergy. Aggregation of FcεRI activates a cascade of signaling events leading to mast cell activation. Subsequently, inhibitory signals are switched on for turning off activated signals. Here we identified that Rcan1 serves as a negative regulator for turning off FcεRI-mediated mast cell activation. FcεRI-induced Rcan1 expression was identified by suppression subtractive hybridization and verified by real-time quantitative PCR and Western blotting. Deficiency of Rcan1 led to increased calcineurin activity, increased NFAT and NF-κB activation, increased cytokine production and enhanced IgE-mediated late phase cutaneous reactions. Forced expression of Rcan1 in wild type or Rcan1 deficient mast cells reduced FcεRI-mediated cytokine production in vitro. Rcan1 deficiency also led to increased FcεRI-mediated mast cell degranulation and enhanced passive cutaneous anaphylaxis. Analysis of the Rcan1 promoter identified a functional Egr1 binding site. Rcan1 expression is preceded by Egr1 expression. Egr1 is a positive regulator of FcεRI-induced cytokine production. Biochemical and genetic evidence suggested that Egr1 controls Rcan1 expression. Our results identified Rcan1 as a novel inhibitory signal in FcεRI-induced mast cell activation and established a new link of Egr1 and Rcan1 in FcεRI signaling.
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25

Baghestanian, Mehrdad, Roland Hofbauer, Hans P. Kiener, Hans C. Bankl, Friedrich Wimazal, Martin Willheim, Otto Scheiner, et al. "The c-kit Ligand Stem Cell Factor and Anti-IgE Promote Expression of Monocyte Chemoattractant Protein-1 in Human Lung Mast Cells." Blood 90, no. 11 (December 1, 1997): 4438–49. http://dx.doi.org/10.1182/blood.v90.11.4438.

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Анотація:
Abstract Recent data suggest that mast cells (MC) are involved in the regulation of leukocyte accumulation in inflammatory reactions. In this study, expression of leukocyte-chemotactic peptides (chemokines) in purified human lung MC (n = 16) and a human mast cell line, HMC-1, was analyzed. Northern blotting and reverse transcriptase-polymerase chain reaction (RT-PCR) showed baseline expression of monocyte chemoattractant protein (MCP)-1 mRNA in unstimulated MC. Exposure of MC to recombinant stem cell factor (rhSCF, 100 ng/mL) or anti-IgE (10 μg/mL) was followed by a substantial increase in expression of MCP-1 mRNA. Neither unstimulated nor stem cell factor (SCF )-stimulated lung MC expressed transcripts for interleukin-8 (IL-8), macrophage inflammatory protein-1α (MIP-1α), MIP-1β, or RANTES by Northern blotting. The mast cell line HMC-1, which contains a mutated and intrinsically activated SCF-receptor, was found to express high levels of MCP-1 mRNA in a constitutive manner. Exposure of HMC-1 cells to rhSCF resulted in upregulation of MCP-1 mRNA expression, and de novo expression of MIP-1β mRNA. The SCF-induced upregulation of MCP-1 mRNA in lung MC and HMC-1 was accompanied by an increase in immunologically detectable MCP-1 in cell supernatants (sup) (lung MC [<98%], control medium, 1 hour: 159 ± 27 v SCF, 100 ng/mL, 1 hour: 398 ± 46 pg/mL/106 cells; HMC-1: control, 1 hour: 894 ± 116 v SCF, 1 hour: 1,536 ± 265 pg/mL/106). IgE-dependent activation was also followed by MCP-1 release from MC. MC-sup and HMC-1–sup induced chemotaxis in blood monocytes (Mo) (control: 100% ± 12% v 2-hour–MC-sup: 463% ± 38% v HMC-1–sup: 532% ± 12%), and a monoclonal antibody (MoAb) to MCP-1 (but not MoAb to IL-8) inhibited Mo-chemotaxis induced by MC-sup or HMC-1–sup (39% to 55% inhibition, P < .05). In summary, our study identifies MCP-1 as the predominant CC-chemokine produced and released in human lung MC. MCP-1 may be a crucial mediator in inflammatory reactions associated with MC activation and accumulation of MCP-1–responsive leukocytes.
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26

Baghestanian, Mehrdad, Roland Hofbauer, Hans P. Kiener, Hans C. Bankl, Friedrich Wimazal, Martin Willheim, Otto Scheiner, et al. "The c-kit Ligand Stem Cell Factor and Anti-IgE Promote Expression of Monocyte Chemoattractant Protein-1 in Human Lung Mast Cells." Blood 90, no. 11 (December 1, 1997): 4438–49. http://dx.doi.org/10.1182/blood.v90.11.4438.4438_4438_4449.

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Анотація:
Recent data suggest that mast cells (MC) are involved in the regulation of leukocyte accumulation in inflammatory reactions. In this study, expression of leukocyte-chemotactic peptides (chemokines) in purified human lung MC (n = 16) and a human mast cell line, HMC-1, was analyzed. Northern blotting and reverse transcriptase-polymerase chain reaction (RT-PCR) showed baseline expression of monocyte chemoattractant protein (MCP)-1 mRNA in unstimulated MC. Exposure of MC to recombinant stem cell factor (rhSCF, 100 ng/mL) or anti-IgE (10 μg/mL) was followed by a substantial increase in expression of MCP-1 mRNA. Neither unstimulated nor stem cell factor (SCF )-stimulated lung MC expressed transcripts for interleukin-8 (IL-8), macrophage inflammatory protein-1α (MIP-1α), MIP-1β, or RANTES by Northern blotting. The mast cell line HMC-1, which contains a mutated and intrinsically activated SCF-receptor, was found to express high levels of MCP-1 mRNA in a constitutive manner. Exposure of HMC-1 cells to rhSCF resulted in upregulation of MCP-1 mRNA expression, and de novo expression of MIP-1β mRNA. The SCF-induced upregulation of MCP-1 mRNA in lung MC and HMC-1 was accompanied by an increase in immunologically detectable MCP-1 in cell supernatants (sup) (lung MC [<98%], control medium, 1 hour: 159 ± 27 v SCF, 100 ng/mL, 1 hour: 398 ± 46 pg/mL/106 cells; HMC-1: control, 1 hour: 894 ± 116 v SCF, 1 hour: 1,536 ± 265 pg/mL/106). IgE-dependent activation was also followed by MCP-1 release from MC. MC-sup and HMC-1–sup induced chemotaxis in blood monocytes (Mo) (control: 100% ± 12% v 2-hour–MC-sup: 463% ± 38% v HMC-1–sup: 532% ± 12%), and a monoclonal antibody (MoAb) to MCP-1 (but not MoAb to IL-8) inhibited Mo-chemotaxis induced by MC-sup or HMC-1–sup (39% to 55% inhibition, P < .05). In summary, our study identifies MCP-1 as the predominant CC-chemokine produced and released in human lung MC. MCP-1 may be a crucial mediator in inflammatory reactions associated with MC activation and accumulation of MCP-1–responsive leukocytes.
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27

Witecka, Joanna, Natalia Malejky-Kłusek, Krzysztof Solarz, Olga Pawełczyk, Małgorzata Kłyś, Aleksandra Izdebska, Weronika Maślanko, and Marek Asman. "The Identification of Potential Immunogenic Antigens in Particular Active Developmental Stages of the Rice Weevil (Sitophilus oryzae)." International Journal of Environmental Research and Public Health 20, no. 5 (February 22, 2023): 3917. http://dx.doi.org/10.3390/ijerph20053917.

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Анотація:
Background: The rice weevil (Sitophilus oryzae) originates from subtropical and tropical areas of Asia and Africa, but it also appears on other continents, mostly as a result of trade in rice. It may occur in grain fields as well as in storage facilities, and cause allergenic reactions. The aim of this study was to identify the potential antigens in all developmental stages of S. oryzae, which may cause an allergic response in humans. Methods: Sera of 30 patients were tested for the presence of IgE antibodies to antigens from three life stages of the rice weevil. To identify protein fractions containing potential allergens, proteins collected from larvae, pupae, and adults separated by sex of S. oryzae were fractionated by SDS-PAGE. Then, they were probed with anti-human, anti-IgE monoclonal antibodies, fractionated by SDS-PAGE and detected by Western blotting. Results: In total, 26 protein fractions of males and 22 fractions of other life stages of S. oryzae (larvae, pupae, and females) positively reacted with the examined sera. Conclusions: The conducted study showed that S. oryzae may be a source of many antigens which may cause the potential allergic reactions in humans.
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28

Mills, F. C., G. Thyphronitis, F. D. Finkelman, and E. E. Max. "Ig mu-epsilon isotype switch in IL-4-treated human B lymphoblastoid cells. Evidence for a sequential switch." Journal of Immunology 149, no. 3 (August 1, 1992): 1075–85. http://dx.doi.org/10.4049/jimmunol.149.3.1075.

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Анотація:
Abstract IgE is produced by B lymphocytes that have undergone a deletional rearrangement of their Ig H chain gene locus, a rearrangement that joins the switch region of the mu gene, S mu, with the corresponding region of the epsilon gene, S epsilon. To examine the resulting composite S mu-S epsilon junctions of human lymphoid cells, we have used a polymerase chain reaction strategy to clone the switch regions of the human myeloma U266 and of two IgE-producing human cell lines generated by treatment of lymphocytes with EBV plus IL-4. The switch junction of one of the EBV lines is a complex rearrangement in which a fragment of S gamma is interposed between S mu and S epsilon. This finding suggested that the switch to epsilon in this human lymphoid cell was preceded by a S mu-S gamma recombination. To determine whether this sequential switch rearrangement represented a unique event or occurred with some regularity in human B cells switching to IgE production, DNA samples from bulk cultures of lymphocytes treated with IL-4 were subjected to polymerase chain reaction amplification of their S mu-S epsilon junctions. When the resulting fragments were examined by Southern blotting, a substantial fraction hybridized to an S gamma probe. This finding suggests that sequential recombination involving S gamma is not rare in the switch to epsilon production in humans. Our polymerase chain reaction strategy should be useful in studying isotype switching at the DNA level.
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29

O'Neill, Geraldine M., and Brian A. Baldo. "Intra-species cross-reactivity of house dust mite allergens separated by protein blotting and detected by selective elution of mite components and IgE antibodies." Electrophoresis 14, no. 1 (1993): 923–25. http://dx.doi.org/10.1002/elps.11501401147.

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30

Shan, E., H. Matsuoka, K. Ando, Y. Chinzei, Y. Taniguchi, Y. Shimizu, and N. Ohtaki. "B-6 Detection of salivary gland antigens of the mosquitoes, Culex pipiens molestus by western blotting analysis with sera of the hypersensitive papients to the mosquito bite." Medical Entomology and Zoology 46, Supplement (1995): 52. http://dx.doi.org/10.7601/mez.46.52_2.

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31

Yang, In Jun, Dong-Ung Lee, and Heung Mook Shin. "Inhibitory Effect of Valencene on the Development of Atopic Dermatitis-Like Skin Lesions in NC/Nga Mice." Evidence-Based Complementary and Alternative Medicine 2016 (2016): 1–11. http://dx.doi.org/10.1155/2016/9370893.

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Анотація:
Valencene (VAL) isolated fromCyperus rotunduspossesses various biological effects such as antiallergic and antimelanogenesis activity. We investigated the effect of VAL on atopic dermatitis (AD) skin lesions and their molecular mechanisms. We topically applied VAL to 1-chloro-2,4-dinitrobenzene (DNCB) sensitized NC/Nga mice. Modified scoring atopic dermatitis index, scratching behavior, and histological/immunohistochemical staining were used to monitor disease severity. RT-PCR, western blotting, and enzyme-linked immunosorbent assay were used to determine the level of IgE, proinflammatory cytokines/chemokines production, and skin barrier proteins expression. Topical application of VAL significantly reduced AD-like symptoms and recovered decreased expression of filaggrin in DNCB-sensitized NC/Nga mice. The levels of serum IgE, IL-1β, IL-6, and IL-13 in skin/splenic tissue were reduced.In vitrostudies using TNF-αand IFN-γtreated HaCaT cells revealed that VAL inhibited the exaggerated expression of Th2 chemokines including TARC/CCL17, MDC/CCL22, and proinflammatory chemokines such as CXCL8, GM-CSF, and I-CAM through blockade of the NF-κB pathway. In addition, expression of the skin barrier protein, involucrin, was also increased by VAL treatment. VAL inhibited the production and expression of proinflammatory cytokines IL-1βand IL-6 in LPS-stimulated RAW 264.7 cells. These results suggest that VAL may serve as a potential therapeutic option for AD.
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32

Zhang, Yongbo, Zhuo Wu, Yihui Yang, and Lu Ding. "Trifluorobenzamidine prevents allergic rhinitis by regulating IgE, IL-4 and IL-5 in T-cells." Tropical Journal of Pharmaceutical Research 19, no. 5 (June 26, 2020): 1023–29. http://dx.doi.org/10.4314/tjpr.v19i5.17.

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Анотація:
Purpose: To investigate the effect of trifluorobenzamidine (TBI) on a mouse model of ovalbumin (OVA)- induced allergic rhinitis. Methods: Allergic rhinitis was established in mice via sensitization on days 1, 5 and 14 through intraperitoneal injection of OVA (100 μg) in PBS. On day 15, the mice were subjected to intranasal exposure to OVA (1.5 mg dissolved in PBS). Prior to 10 days of intranasal exposure to OVA, the micewere treated with TBI at doses of 5, 10 and 20 μg/kg. Cytokine levels were determined using enzymelinked immunosorbent assay (ELISA) kits, while cyclooxygenase (COX)-2 and caspase-1 activity were assayed with western blotting. Results: Treatment with TBI significantly (p < 0.05) reduced OVA-mediated increases in nasal rub scores, and decreased serum levels of IgE, TNF-α, thymic stromal lymphopoietin (TSLP), IL-1β and histamine in mice. It also significantly regulated spleen weight and IL-4 secretion (p < 0.05) in OVAadministered mice. TBI significantly downregulated the expressions of IL-5, IL-13, TNFα, TSLP, IL-1β and IL-6 (p < 0.05). Administration of TBI caused a marked reduction in OVA-mediated increase in caspase-1 activity in mice intranasal tissues, and also significantly reduced OVA-induced excessive production of MIP-2 and ICAM-1 (p < 0.05). Moreover, TBI prevented OVA-induced infiltration of eosinophils and mast cells into intranasal tissues (p < 0.05). Conclusion: TBI reduces levels of IgE and various pro-inflammatory cytokines in OVA-administered mice. It also regulates Th1:Th2 ratio, inhibited activity of caspase-1, suppressed mast cell/eosinophil infiltration and reduced ICAM-1 and MIP-2 levels. Therefore, TBI possesses inhibitory potential against rhinitis allergy, and thus can potentially be developed as a new treatment strategy for asthma. Keywords: Trifluorobenzamidine, Anti-inflammation, Allergic rhinitis, Cytokines, Caspase-1, Itching
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Bai, Donghui, Tianxiao Sun, Fang Lu, Yancheng Shen, Yan Zhang, Bo Zhang, Guangli Yu, Haihua Li та Jiejie Hao. "Eupatilin Suppresses OVA-Induced Asthma by Inhibiting NF-κB and MAPK and Activating Nrf2 Signaling Pathways in Mice". International Journal of Molecular Sciences 23, № 3 (29 січня 2022): 1582. http://dx.doi.org/10.3390/ijms23031582.

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Анотація:
To investigate the effect of eupatilin in asthma treatment, we evaluated its therapeutic effect and related signal transduction in OVA-induced asthmatic mice and LPS-stimulated RAW264.7 cells. The BALF was tested for changes in lung inflammatory cells. Th2 cytokines in the BALF and OVA-IgE in the serum were measured by ELISA. H&E and PAS staining were used to evaluate histopathological changes in mouse lungs. The key proteins NF-κB, MAPK, and Nrf2 in lung tissues were quantitatively analyzed by Western blotting. Finally, we evaluated the effect of eupatilin on cytokines and related protein expression in LPS-stimulated RAW 264.7 cells in vitro. In OVA-induced asthmatic mice, eupatilin reduced the numbers of inflammatory cells, especially neutrophils and eosinophils. Eupatilin also decreased the levels of IL-5, IL-13 in the BALF and OVA-IgE in the serum. Furthermore, eupatilin inhibited the activation of NF-κB and MAPK pathways and increased the expression of Nrf2 in OVA-induced asthmatic mice. In vitro, eupatilin significantly reduced LPS-stimulated NO, IL-6, and ROS production. Additionally, the NF-κB, MAPK, and Nrf2 protein expression in LPS-stimulated RAW264.7 cells was consistent with that in OVA-induced asthmatic lung tissues. In summary, eupatilin attenuated OVA-induced asthma by regulating NF-κB, MAPK, and Nrf2 signaling pathways. These results suggest the utility of eupatilin as an anti-inflammatory drug for asthma treatment.
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34

Lin, Tzou-Yien, Tsong-Min Chang, and Huey-Chun Huang. "Extracellular Vesicles Derived from Human Umbilical Cord Mesenchymal Stem Cells Attenuate Mast Cell Activation." Antioxidants 11, no. 11 (November 17, 2022): 2279. http://dx.doi.org/10.3390/antiox11112279.

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Анотація:
The therapeutic potential of extracellular vesicles isolated from stem cells have been reported in several clinical diseases. Preclinical studies have demonstrated the beneficial effects of extracellular vesicles in the treatment of heart, kidney, liver, brain, and skin injuries. To address the putative therapeutic effects and mechanisms of extracellular vesicles derived from human umbilical cord mesenchymal stem cells on allergic activation in mast cells, we isolated extracellular vesicles from human umbilical cord-derived mesenchymal stem cells (UCMSCs) by tangential-flow filtration methods. The characteristics and identification of UCMSC-derived extracellular vesicles were examined via nanoparticle tracking analysis, transmission electron microscopy and protein marker analysis. Cytokines and tryptase in the cultured supernatant of KU812 cells were analyzed using an ELISA kit. Proteins in the MAPK and STAT5 signaling pathways were detected by Western blotting. This study showed that different doses of UCMSC-derived extracellular vesicles abolish IgE-stimulated KU812 cell activation and reduce the level of NF-κB, which subsequently leads to cell degranulation and the release of IL-1β, TNF-α and IL-6. Additionally, UCMSC-derived extracellular vesicles treatment blunted the IgE-induced signaling proteins p-P38, p-JNK and p-STAT5. Our results revealed a mechanism for anti-inflammation in which extracellular vesicles can affect the activation of mast cells and thus function in allergy regulation.
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35

Ma, Wenli, Xiaojuan Ma, Xiaofan Li, Xuebin Xi, Chunyang Mao, and Lixin Wang. "The Effect and Mechanism of Burnet Gels on Steroid-Dependent Dermatitis in Guinea Pig Model." BioMed Research International 2022 (September 13, 2022): 1–9. http://dx.doi.org/10.1155/2022/5866824.

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Анотація:
Objective. This study was designed to establish quality standards of Burnet gels and investigate the effects and mechanism of Burnet gels on steroid-dependent dermatitis (HDD) in guinea pigs. Methods. HPLC was used to determine the content of gallic acid, Gentiopicrin, and paeonol. A total of 48 male guinea pigs were recruited and randomly divided into control group, model group, tacrolimus ointment group, and Burnet gel group (Low, medium, and high concentration). The HDD guinea pig model was established by the 0.5% clobetasol propionate tincture. After HDD model establishment, control group and model group smeared normal saline and the rest of the group with corresponding drugs for three weeks. The contents of IFN-γ, IL-4, and IgE in the guinea pig serum were detected by the ELISA; the protein expression levels of FLG, LOR, and Caspase-14 in the epidermis of guinea pigs were detected by the immunohistochemical and Western blotting method. Results. The content of gallic acid, Gentiopicrin, and paeonol was 0.30 mg/g, 1.06 mg/g, and 0.56 mg/g. Compared with the normal group, the IFN-γ, IL-4, and IgE of guinea pig serum in the model group were significantly increased; the FLG, LOR, and Caspase-14 of guinea pig epidermis in the model group were significantly decreased; compared with the model group, the IFN-γ, IL-4, and IgE of guinea pig serum in the tacrolimus ointment group and Burnet gel group were significantly decreased; the FLG, LOR, and Caspase-14 of guinea pig epidermis in the tacrolimus ointment group and Burnet gel group were significantly increased. Conclusion. Burnet gels can improve guinea pig HDD model, and the mechanism may be related to inhibiting skin inflammation and promoting the formation of epidermal skin barrier.
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36

Wang, Yajuan, Huizhi Zhu, Jiabing Tong, and Zegeng Li. "Ligustrazine Inhibits Lung Phosphodiesterase Activity in a Rat Model of Allergic Asthma." Computational and Mathematical Methods in Medicine 2022 (January 10, 2022): 1–10. http://dx.doi.org/10.1155/2022/1452116.

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Анотація:
Objectives. This study sought to examine whether ligustrazine was capable of inhibiting phosphodiesterase (PDE) activity and improving lung function in a rat model of asthma. Methods. Rats were initially sensitized using ovalbumin (OVA) and then were challenged daily with aerosolized OVA beginning 14 days later (30 min/day) to generate a rat model of asthma. Changes in airway function following methacholine (MCh) injection were evaluated by monitoring lung resistance ( R L ) and dynamic lung compliance ( C dyn ) values using an AniRes2005 analytic system. In addition, serum IgE was measured via ELISA, while PDE expression was evaluated via qPCR and western blotting. Key Findings. Ligustrazine significantly impaired allergen-induced lung hyperresponsivity and inflammation in this asthma model system. Ligustrazine treatment was also associated with reduced expression of PDEs including PDE4 in the lungs of these rats. Conclusions. Ligustrazine suppresses airway inflammation and bronchial hyperresponsivity in this rat model system, and these changes are associated with decreased PDE expression at the protein and mRNA levels.
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37

Stavnezer, J., S. Sirlin, and J. Abbott. "Induction of immunoglobulin isotype switching in cultured I.29 B lymphoma cells. Characterization of the accompanying rearrangements of heavy chain genes." Journal of Experimental Medicine 161, no. 3 (March 1, 1985): 577–601. http://dx.doi.org/10.1084/jem.161.3.577.

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Анотація:
The murine B cell lymphoma I.29 contains cells expressing surface IgM or IgA with identical heavy chain variable regions (9, 25, and D. Klein and J. Stavnezer, unpublished data). Purified IgM+ cells from the lymphoma have been adapted to culture and induced to switch to IgA, IgE, or IgG2 by treatment with lipopolysaccharide (LPS) or by treatment with a monoclonal anti-I.29 antiidiotype plus LPS. Clones of IgM+ cells have been obtained and induced to switch. Under optimal conditions, 30% of the cells in the culture expressed IgA 8 d after the inducers were added, and by 15 d 90% of the cells were IgA+. In actively switching cultures, up to 50% of the cells whose cytoplasm stained positively with anti-IgA stained simultaneously with anti-IgM, which indicates that the appearance of IgA+ cells in the cultures was due to isotype switching and not to clonal outgrowth. Examination by Southern blotting experiments of the Ig heavy chain genes in I.29 cells before and after switching revealed that isotype switching was accompanied by DNA recombinations that occurred within or immediately 5' to the tandemly repeated switch sequences. Within 3 d after the addition of inducers of switching, the nonexpressed chromosome underwent a variety of deletions or expansions within the S mu region, and a portion of the S alpha regions had undergone a 0.9-kb deletion. In cultures that contained at least 12% IgA+ cells, rearranged, expressed alpha genes, produced by recombination between the S mu region within the expressed mu gene and the S alpha region, were detected.
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38

AMANO, Maho, Haruko OGAWA, Kyoko KOJIMA, Tomoko KAMIDAIRA, Susumu SUETSUGU, Munehiro YOSHIHAMA, Takanori SATOH, Tatsuya SAMEJIMA, and Isamu MATSUMOTO. "Identification of the major allergens in wheat flour responsible for baker's asthma." Biochemical Journal 330, no. 3 (March 15, 1998): 1229–34. http://dx.doi.org/10.1042/bj3301229.

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Анотація:
Baker's asthma, a typical occupational allergic disease, is a serious problem in the food industries. In this study, purification and identification of major allergens recognized by IgEs in sera of allergic patients were performed. Major immunoreactive proteins were purified from the albumin fraction by gel filtration on a Toyopearl HW-50 column followed by reverse-phase HPLC. The N-terminal amino acid sequences and molecular masses measured by MS indicated that the major immunoreactive proteins are members of the α-amylase inhibitor family, 0.19 and 0.28. Significant leukotriene release by each purified protein was observed in cell-associated stimulation tests, suggesting in vivo activity of these antigens. Carbohydrate analyses of major allergens indicated that they are monoglycosylated but not N-glycosylated in spite of the presence of a potential N-glycosylation site. Recombinant 0.19 expressed in Escherichia coli showed the same reactivity with IgE as native wheat 0.19 in Western blotting and ELISA using methyl vinyl ether maleic anhydride co-polymer as an immobilizing reagent, suggesting that the allergenic epitopes are located in the peptide portions.
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39

Fukuoka, Yoshihiro, Michelle R. Hite, and Lawrence B. Schwartz. "Functional angiotensin and bradykinin receptors are expressed on primary human mast cells." Journal of Immunology 196, no. 1_Supplement (May 1, 2016): 126.19. http://dx.doi.org/10.4049/jimmunol.196.supp.126.19.

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Abstract Purpose The role of mast cells (MCs) has traditionally been recognized as an effector cell for IgE-mediated allergic diseases, involving ~20% of USA citizens. However, everyone has MCs, indicating their involvement in biologic and pathobiologic conditions beyond allergy. Angiotensin converting enzyme inhibitors (ACEI) and angiotensin II type 1 receptor (AT1R) blockers are used to treat cardiovascular disorders such as hypertension, while a bradykinin type 2 receptor (B2R) inhibitor is used to treat attacks of hereditary angioedema. We examined the expression and biological function of AT1R, B2R and B1R on primary human MCs. Methods MCs were dispersed from fresh surgical human skin obtained from CHTN, and placed into culture with SCF (100 ng/ml) for 4–8 weeks. FACS analysis, western blotting, qRT-PCR, degranulation (β-hexosaminidase release), cytokine/chemokine secretion of IL-6, IL-8 and CCL5, nitric oxide (NO) production (DAF-AM diacetate) were measured. Results FACS analysis and western blotting showed the AT1R and B2R are expressed on MCs. Angiotensin II (0.1–10 μM) and bradykinin (BK, 0.1–2 μM) stimulated MCs to release cytokines (1–4 ng/ml) which was inhibited by specific inhibitors. BK also stimulated the production of NO. Both the B2R and B1R are up-regulated by TNFα (10 ng/ml for 3 days), leading to enhanced activation of MCs by BK (1 μM) and desArg-LysBK (1 μM). Exposure of MCs to angiotensin II (2 μM), angiotensin (1–7)(10 μM) or ACEI (ramipril 1 μM) also enhanced cytokine secretion triggered by low concentration of BK (0.03–0.25 μM). Conclusion These findings suggest functional interactions of AT1R, B2R, ACE and Mas on human MCs, providing novel roles for MCs in cardiovascular, inflammatory and allergic disorders.
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40

Wang, Sang, Lei Wang, Hua Hu, and Pin Dong. "MiR-224 ameliorates inflammation and symptoms in mouse model of allergic rhinitis by targeting CDK9." Allergologia et Immunopathologia 49, no. 6 (November 5, 2021): 80–88. http://dx.doi.org/10.15586/aei.v49i6.451.

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Objectives To explore the regulatory effects of microRNA (miRNA)-224 and its potential target gene, cyclin dependent kinase 9 (CDK9), in the pathological process of allergic rhinitis (AR). Methods To investigate the role of miR-224 and CDK9, it was screened by bioinformatics prediction software and verified by dual-luciferase reporter assay. The mouse model of AR was established by ovalbumin (OVA).The animal models were intervened with miR-224 agomir, negative control agomir, and saline respectively. The symptoms of sneezing and nasal rubbing were recorded. The expressions of miR224, CDK9, and cytokines in the nasal mucosa of different groups were analyzed by rt-PCR or western blotting. Enzyme-linked immunoassay (ELISA) was used to evaluate the levels of IgE and Histamine (HA) in the serum. The infiltration of inflammatory cells in the nasal mucosa was studied by immunohistochemistry. The expression and distribution of CDK9 in the nasal mucosa of mice were revealed by immunofluorescence. Results In the nasal mucosa of the animal models, the level of miR-224 was downregulated, while that of CDK9 was upregulated. The upregulation of miR-224 by miR-224 agomir reduced the frequencies of nasal rubbing and sneezing, the expression of CDK9, the levels of cytokines, and the concentrations of IgE and HA. Moreover, miR-224 appeared to attenuate the infiltration of inflammatory cells and hypersecretion of glands in the nasal mucosa. The expression of CDK9, which was distributed under the mucosa, especially in the submucosa interstitial tissue, was significantly reduced. Conclusion MiR-224 affected the pathogenesis of AR by targeting CDK9. It proves that miR-224 could be a novel potential therapeutic target for AR.
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41

Yang, Yong Jun, Wei Chen, Alexander Edgar, Bo Li, Jeffery D. Molkentin, Jason N. Berman та Tong-Jun Lin. "Rcan1 negatively regulates FcɛRI-mediated signaling and mast cell function". Journal of Experimental Medicine 206, № 1 (5 січня 2009): 195–207. http://dx.doi.org/10.1084/jem.20081140.

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Анотація:
Aggregation of the high affinity IgE receptor (FcɛRI) activates a cascade of signaling events leading to mast cell activation. Subsequently, inhibitory signals are engaged for turning off activating signals. We identified that regulator of calcineurin (Rcan) 1 serves as a negative regulator for turning off FcɛRI-mediated mast cell activation. FcɛRI-induced Rcan1 expression was identified by suppression subtractive hybridization and verified by real-time quantitative polymerase chain reaction and Western blotting. Deficiency of Rcan1 led to increased calcineurin activity, increased nuclear factor of activated T cells and nuclear factor κB activation, increased cytokine production, and enhanced immunoglobulin E–mediated late-phase cutaneous reactions. Forced expression of Rcan1 in wild-type or Rcan1-deficient mast cells reduced FcɛRI-mediated cytokine production. Rcan1 deficiency also led to increased FcɛRI-mediated mast cell degranulation and enhanced passive cutaneous anaphylaxis. Analysis of the Rcan1 promoter identified a functional Egr1 binding site. Biochemical and genetic evidence suggested that Egr1 controls Rcan1 expression. Our results identified Rcan1 as a novel inhibitory signal in FcɛRI-induced mast cell activation and established a new link of Egr1 and Rcan1 in FcɛRI signaling.
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42

Wolff, N., U. Cogan, H. Zuckerman, N. Karin, Y. Levy, Y. E Krasik, J. Felsteiner, R. Reifen, and S. Yannai. "Decrease of the allergenic activity of foods by shock waves." Czech Journal of Food Sciences 22, SI - Chem. Reactions in Foods V (January 1, 2004): S36—S39. http://dx.doi.org/10.17221/10607-cjfs.

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Анотація:
Food allergy significantly affects the life-quality for many people worldwide and is life-threatening in extreme cases. In recent years the incidence of this disease shows a gradual increase in many countries. It is a well-established fact that common food-processing operations involving heat treatments fail to significantly decrease the allergenic reactivity of foods. Furthermore, allergenic proteins are remarkably resistant to proteolysis by digestive enzymes and often remain intact after passing through the gastrointestinal tract. The tested materials were protein extracts from sesame seeds, milk and peanuts, or isolated proteins from the same sources. Treatments investigated included application of 6 to 10 pulses of high-frequency acoustic shock waves, lasting a few microseconds. The treated samples were tested in vitro by Western blotting with sera from humans diagnosed to be allergic to the food in question, and in vivo by measuring the IgE levels produced in young Brown Norway rats exposed to the tested proteins by sensitisation through i.p. injection, or by feeding for up to 6 weeks, using direct and indirect ELISA. The treatments markedly decreased or completely eliminated the allergenic reactivity of the foods, as evidenced by the assays used.
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43

Mozyrska, O. V. "The tol-like receptor 2 polymorphism significance for the development of sensitization to house dust mites in children with atopic dermatitis." Modern pediatrics. Ukraine, no. 4(124) (May 30, 2022): 60–64. http://dx.doi.org/10.15574/sp.2022.124.60.

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Анотація:
Atopic dermatitis is a chronic recurrent inflammatory skin disease that affects 5-20% of children. Airborne allergens derived from house dust mites can cause atopic dermatitis. TLR2 play an important role in the recognition of house dust mite allergens. Purpose - to investigate the prevalence of sensitization to house dust mites in children with atopic dermatitis and the role of TLR2 rs4696480 polymorphism in the development of sensitivity to house dust mites. Materials and methods. The study included 100 patients with atopic dermatitis. Genotyping of the polymorphism TLR2 rs4696480 was performed in the patient group using real-time PCR. Measurements of sIgE to dust mites were performed by Western blotting according to the manufacturer’s protocol (Simesta-Medivis, Ukraine-Germany). Results. Sensitization to house dust mites was found in 48% of children. Children with elevated levels of specific IgE to dust mites had a significantly higher SCORAD index than patients without sensitization (p<0.001). In the group of children sensitized to house dust mites, there were significantly higher levels of total IgE (p<0.001) and a longer course of the disease (p<0.05). There was no statistically significant difference in the distribution of genotypes depending on the presence of sensitization to dust mites (OR=1.250 (0.481-3.245) for AA and AT; OR=2.125 (0.715-6.315) for AA and TT). Conclusions. This study showed that the susceptibility to dust mites among children with atopic dermatitis is 48%. The presence of susceptibility to house dust mites affects the severity of the disease. The research was carried out in accordance with the principles of the Helsinki Declaration. The study protocol was approved by the Local ethics committee of the participating institution. The informed consent of the patient was obtained for conducting the studies. No conflict of interests was declared by the author. Key words: TLR2, house dust mites, atopic dermatitis, children.
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44

Premrov Bajuk, Blanka, Petra Zrimšek, Tina Kotnik, Adrijana Leonardi, Igor Križaj, and Breda Jakovac Strajn. "Insect Protein-Based Diet as Potential Risk of Allergy in Dogs." Animals 11, no. 7 (June 29, 2021): 1942. http://dx.doi.org/10.3390/ani11071942.

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Анотація:
Before insects can be used widely as an alternative source of dietary protein, their allerginicity should be investigated. Therefore, the aim of our study was to assess the potential adverse reactions of the immune system of dogs against Tenebrio molitor proteins. Dogs sensitised to storage mites T. putrescentiae and A. siro were included. Clinically healthy and clinically allergic dogs were compared. Proteins were extracted from mealworm larvae and their digestibility determined by in vitro incubation with digestive proteases. Mealworm protein extracts and digests were analysed by SDS–PAGE. Canine sera tested for the presence of mite-specific IgEs were used for subsequent Western blotting. LC-MS/MS analysis was used to identify mealworm proteins and their allergenic potential was predicted with the AllermatchTM tool. The binding of canine sera IgEs to mealworm proteins was confirmed; however, the differences between the two groups of dogs were not significant. Moreover, no clear correlation was found between sensitisation to storage mites and clinical status of the dogs. Altogether, 17 different proteins were identified, including tropomyosin, α-amylase, and Tm-E1a cuticular protein that are known cross-reacting IgE-binding allergens. Our results suggest that dogs allergic to mites may clinically express also the cross-reactivity with mealworm proteins.
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45

Kulmburg, P. A., N. E. Huber, B. J. Scheer, M. Wrann, and T. Baumruker. "Immunoglobulin E plus antigen challenge induces a novel intercrine/chemokine in mouse mast cells." Journal of Experimental Medicine 176, no. 6 (December 1, 1992): 1773–78. http://dx.doi.org/10.1084/jem.176.6.1773.

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Анотація:
In an attempt to characterize genes participating in the allergic late phase reaction, we have isolated a novel intercrine/chemokine (called MARC) from a cDNA library of the stimulated mouse mast cell line, CPII. As measured by Northern blotting, it is strongly upregulated at the mRNA level after the physiological challenge of the cells with immunoglobulin (Ig)E plus antigen. Unstimulated cells completely lack significant, stable expression, as do a number of other, different cell lines (uninduced and induced) and mouse tissues. In contrast to the Northern blot analysis, a polymerase chain reaction (PCR) analysis, performed on CPII cells and on Percoll gradient purified mouse peritoneal mast cells, revealed a basal level of transcription in the uninduced stage. After 2 h of IgE plus antigen challenge, a quantitative reverse transcriptase-PCR, using a spiked in MIMIC, showed a level of transcripts more than 100-fold higher in the CPII cells and 5-20-fold higher in purified mouse peritoneal cavity mast cells. This rapid induction after the Fc epsilon RI challenge, the identification of the gene as a member of the chemokine family, and its upregulated expression in peritoneal mast cells, all suggest an involvement in certain acute and chronic pathological mast cell-driven diseases.
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46

Đukić, Teodora, Katarina Smiljanić, Jelena Mihailović, Ivana Prodić, Danijela Apostolović, Shu-Hua Liu, Michelle M. Epstein, Marianne van Hage, Dragana Stanić-Vučinić, and Tanja Ćirković Veličković. "Proteomic Profiling of Major Peanut Allergens and Their Post-Translational Modifications Affected by Roasting." Foods 11, no. 24 (December 9, 2022): 3993. http://dx.doi.org/10.3390/foods11243993.

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Post-translational modifications (PTMs) are covalent changes occurring on amino acid side chains of proteins and yet are neglected structural and functional aspects of protein architecture. The objective was to detect differences in PTM profiles that take place after roasting using open PTM search. We conducted a bottom-up proteomic study to investigate the impact of peanut roasting on readily soluble allergens and their PTM profiles. Proteomic PTM profiling of certain modifications was confirmed by Western blotting with a series of PTM-specific antibodies. In addition to inducing protein aggregation and denaturation, roasting may facilitate change in their PTM pattern and relative profiling. We have shown that Ara h 1 is the most modified major allergen in both samples in terms of modification versatility and extent. The most frequent PTM was methionine oxidation, especially in roasted samples. PTMs uniquely found in roasted samples were hydroxylation (Trp), formylation (Arg/Lys), and oxidation or hydroxylation (Asn). Raw and roasted peanut extracts did not differ in the binding of IgE from the serum of peanut-sensitised individuals done by ELISA. This study provides a better understanding of how roasting impacts the PTM profile of major peanut allergens and provides a good foundation for further exploration of PTMs.
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47

Knol, E. F., M. Wensing, R. Vlooswijk, M. Ertmann, A. C. Knulst, and S. J. Koppelman. "Relevance of ara h1, ara h2, and ara h3 in peanut allergic patients, as determined by IgE-western-blotting, basophil histamine release, and intracultaneous testing: Ara h2 is the most important peanut allergen." Journal of Allergy and Clinical Immunology 111, no. 2 (February 2003): S194—S195. http://dx.doi.org/10.1016/s0091-6749(03)80665-6.

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48

Yuan, Yamei, Xiangming Fang, and Weidong Ye. "Acrid and Bitter Chinese Herbs in Decoction Effectively Relieve Lung Inflammation and Regulation of TRPV1/TAS2R14 Channels in a Rat Asthmatic Model." Evidence-Based Complementary and Alternative Medicine 2022 (August 22, 2022): 1–10. http://dx.doi.org/10.1155/2022/8061740.

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Анотація:
Background. Shegan Mahuang decoction (SGMHD) was widely used as a classic prescription of traditional Chinese medicine to treat asthma. However, there is no research on the acrid and bitter Chinese herbs in the SGMHD to treat asthma. This study aimed to investigate the effects of SGMHD and its acrid-bitter Chinese herbs composition on airway inflammation and the expression of TRPV1 and TAS2R14 genes and proteins in asthmatic rats. Methods. SD (Sprague Dawley) rats of asthma were induced by ovalbumin and aluminum hydroxide, then randomly divided into the Normal group, Model group, SGMHD group, Dexamethasone (Dex) group, Guilongkechuangning (GLKC) group, The Acrid Chinese Herbs group (ACH), and The Bitter Chinese Herbs group (BCH). The rats were given intragastric gavage after 21 days for 4 weeks. The bronchoalveolar lavage fluid (BALF) was collected, and the levels of IL-4, IL-13, nerve factors SP, CGRP, PGE2, and serum of IgE were determined by ELISA. Pathological changes in the lungs were determined by hematoxylin-eosin (HE) staining. The expression of TRPV1 and TAS2R14 in the rat lung group was detected by immunofluorescence (IF). The expression levels of TRPV1 and TAS2R14 were measured using western blotting. The mRNA levels of TRPV1 and TAS2R14 were measured using RT-qPCR. Results. The levels of serum IgE in treated rats and the cytokines IL-4, IL-13, SP, CGRP, and PGE2 were all decreased. HE-staining showed that significantly reduced inflammatory cell infiltration in lung tissue. IF-staining showed the expression levels except those of the normal group were enhanced. Acrid Chinese herbs inhibited TRPV1, and bitter Chinese herbs activated the gene and protein expression of TAS2R in the lung. Conclusion. The acrid Chinese herbs regulate TRPV1, and bitter Chinese herbs regulate the gene and protein expression of TAS2R14, through nerve and immune-inflammatory factors, reduced airway inflammation, reduced airway reactivity, promoted airway remodeling, and the combination of acrid-bitter Chinese herbs can enhance the above effects. This will lay a foundation for further in vivo study of specific compounds of acrid-bitter Chinese herbs.
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49

Aye, Cho Cho, and Santa Ono. "Chemokine receptor signaling in mast cells (89.27)." Journal of Immunology 178, no. 1_Supplement (April 1, 2007): S154. http://dx.doi.org/10.4049/jimmunol.178.supp.89.27.

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Abstract Mast cells play an important role in IgE-associated allergic disorders and immune response to parasites. In a former study, we demonstrated that chemokine CCL3 (MIP-1α) acted as a co-stimulator for FcεRI-mediated activation on mast cells. To advance our understanding on this finding, we studied gene profiles in Rat Basophilic Leukaemia mast cells after co-activation of CCR1 (a receptor of CCL3) and FcεRI using the Affymetrix GeneChip Arrays. Emp1 (Epithelial Membrane protein 1), RT1-S3 (non-classical type 1 MHC gene), and Slc35e4 (solute carrier family 35, member E4) were found to be the top possible synergy-related genes. Validations of this data are being carried out using real time PCR, western-blotting and siRNA technologies. Further to our in vivo findings of the inhibition of the immediate hypersensitivity reactions in the conjunctiva and the attenuation of mast cell degranulation in MIP-1α deficient mice, we would like to check if MIP-1α and CCR1 play a critical role to produce these effects. We reconstituted the bone marrow-derived mice mast cells (BMMC) from naïve mice into mast cell-deficient mice and confirmed that the reconstituted BMMC home to their eye. The next step is to examine whether CCR1 and CCL3 are critical for mast cell activation and allergic conjunctivitis-like symptoms following induction of murine allergic conjunctivitis using CCR1- and CCL3- deficient mice.
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50

Liang, Xiao, Chang-Shun Liu, Xiao-Han Wei, Ting Xia, Fei-Long Chen, Qing-Fa Tang, Meng-Yue Ren, and Xiao-Mei Tan. "Mahuang Fuzi Xixin Decoction Ameliorates Allergic Rhinitis in Rats by Regulating the Gut Microbiota and Th17/Treg Balance." Journal of Immunology Research 2020 (May 25, 2020): 1–11. http://dx.doi.org/10.1155/2020/6841078.

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Анотація:
Mahuang Fuzi Xixin Decoction (MFXD), a Chinese traditional herbal formulation, has been used to treat allergic rhinitis (AR) in China for centuries. However, the mechanism underlying its effect on AR is unclear. This study investigated the mechanism underlying the therapeutic effects of MFXD on AR. Ovalbumin-induced AR rat models were established, which were then treated with MFXD for 14 days. Symptom scores of AR were calculated. The structure of the gut microbiota was analyzed by 16S rRNA gene sequencing and qPCR. Short-chain fatty acid (SCFA) content in rat stool and serum was determined by GC-MS. Inflammatory and immunological responses were assessed by histopathology, ELISA, flow cytometry, and western blotting. Our study demonstrated that MFXD reduced the symptom scores of AR and serum IgE and histamine levels. MFXD treatment restored the diversity of the gut microbiota: it increased the abundance of Firmicutes and Bacteroidetes and decreased the abundance of Proteobacteria and Cyanobacteria. MFXD treatment also increased SCFA content, including that of acetate, propionate, and butyrate. Additionally, MFXD administration downregulated the number of Th17 cells and the levels of the Th17-related cytokines IL-17 and RORγt. By contrast, there was an increase in the number of Treg cells and the levels of the Treg-related cytokines IL-10 and Foxp3. MFXD and butyrate increased the levels of ZO-1 in the colon. This study indicated MFXD exerts therapeutic effects against AR, possibly by regulating the gut microbial composition and Th17/Treg balance.
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