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Laccase/caffeic acid-catalyzed crosslinking coupled with galactomannan alters the conformational structure of ovalbumin and alleviates Th2-mediated allergic asthma

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Food Science and Human Wellness 2024年第4期13卷 1962-1973页

作者： Ishfaq Ahmed Suidong Ouyang Shengquan Wu Haochang Song Miaoyuan Zhang Renxing Luo Peishan Lu Jiaqi Deng Tingting Zheng Yanyan Wang Xinguang Liu Gonghua Huang Guangdong Provincial Key Laboratory of Medical Molecular Diagnostics The First Dongguan Affiliated HospitalGuangdong Medical UniversityDongguan 523808China Haide College Ocean University of ChinaLaoshan CampusQingdao 266100China School of Medical Technology Guangdong Medical UniversityDongguan 523808China

ovalbumin(OVA)is the major allergenic protein that can induce T helper 2(Th2)-allergic reactions,for which current treatment options are inadequate.In this study,we developed a polymerized hypoallergenic OVA product via laccase/caffeic acid(Lac/CA)-catalyzed crosslinking in conjunction with galactomannan(Man).The formation of high molecular weight crosslinked polymers and the Ig G-binding were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)and Western blotting.The study indicated that Lac/CA-catalyzed crosslinking plus Man conjugation substantially altered secondary and tertiary structures of OVA along with the variation in surface hydrophobicity.Gastrointestinal digestion stability assay indicated that crosslinked OVA exhibited less resistance in simulated gastric fluid(SGF)and simulated intestinal fluid(SIF).Mouse model study indicated that Lac-Man/OVA ameliorated eosinophilic airway inflammatory response and efficiently downregulated the expression of Th2-related cytokines(interleukin(IL)-4,IL-5,and IL-13),and upregulated IFN-γand IL-10 expression.Stimulation of bone marrow-derived dendritic cells with Lac-Man/OVA suppressed the expression of phenotypic maturation markers(CD80 and CD86)and MHC class II molecules,and suppressed the expression levels of proinflammatory cytokines.The knowledge obtained in the present study offers an effective way to acquire a hypoallergenic OVA product that can have a therapeutic effect in alleviating OVA-induced allergic asthma.

关键词： ovalbumin Laccase Galactomannan Conformational structure Asthma

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Rosmarinic acid improves tracheal smooth muscle responsiveness and lung pathological changes in ovalbumin-sensitized rats

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Asian Pacific Journal of Tropical Biomedicine 2024年第3期14卷 100-105页

作者： Vahideh Abbasnia Mohsen Foadoddini Delaram Eslimi Esfahani Mohammad Reza Khazdair Shahrbanoo Oryan Department of Biology Payame Noor UniversityTehranIran Cardiovascular Diseases Research Center Birjand University of Medical SciencesBirjandIran Department of Animal Sciences Faculty of Biological SciencesKharazmi UniversityTehranIran

Objective:To evaluate the effect of rosmarinic acid on tracheal smooth muscle responsiveness and lung pathological changes in ovalbumin-sensitized rats.Methods:Rats were randomly divided into six groups:the control group,the asthmatic group,and the asthmatic groups treated with dexamethasone(1 mg/kg;oral gavage)or three doses of rosmarinic acid(0.5,1,and 2 mg/kg;oral gavage).For induction of asthma,rats received intraperitoneal injections and inhalation of ovalbumin.After 21 days,bronchoalveolar lavage fluid and lung samples were collected for histopathological analyses.Moreover,total and differential white blood cell counts were determined.Results:The rosmarinic acid-treated group had significantly lower tracheal smooth muscle responses to methacholine than the asthmatic group.In addition,rosmarinic acid reduced white blood cell count and the percentages of eosinophils,monocytes,and neutrophils while increasing the percentage of lymphocytes.ovalbumin-induced lung pathological changes were significantly improved by treatment with rosmarinic acid.Conclusions:Rosmarinic acid improves tracheal smooth muscle responsiveness and lung pathological changes in ovalbumin-sensitized rats.

关键词： Rosmarinic acid Asthma ovalbumin Tracheal responsiveness Inflammation White blood cell

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Boeravinone B ameliorates allergic nasal inflammation by modulating the GATA-3/T-bet signaling pathway in a mouse model of allergic rhinitis

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Asian Pacific Journal of Tropical Biomedicine 2024年第6期14卷 245-252页

作者： Ben Ye Ji-Zhe Wang Department of Otorhinolaryngology Second Hospital of Dalian Medical UniversityDalian116000China Department of Otorhinolaryngology Shandong Provincial Hospital Affiliated to Shandong First Medical UniversityJinan250000China

Objective:To evaluate the anti-allergic effect of boeravinone B against ovalbumin-induced allergic rhinitis in mice and explore its possible mechanism.Methods:For the induction of allergic rhinitis,mice were intraperitoneally sensitized and intranasally challenged with ovalbumin,as well as orally received various concentrations of boeravinone B.Nasal mucosal inflammation,and the levels of nitric oxide,β-hexosaminidase,IFN-γ,LTC-4,myeloperoxidase,Nrf2,HO-1,GATA-3,ROR-γ,T-bet,antioxidant parameters,and allergen-specific cytokines were assessed.Results:Boeravinone B markedly reduced ovalbumin-induced increase in the number of episodes of nasal sneezing,rubbing,and discharge,as well as the levels of IgE,IgG1,andβ-hexosaminidase(P<0.05).It also significantly reduced differential cell count,myeloperoxidase,oxide-nitrosative stress,and the levels of IL-1β,IL-4,IL-5,IL-6,IL-13,IL-17,tumor necrosis factor-α,GATA-3,and ROR-γwhile enhancing the level of T-bet.Conclusions:Boeravinone B is a potential therapeutic agent for allergic rhinitis by modulating various inflammatory mediators and immune responses.

关键词： Allergic reaction Boeravinone B Oxidative stress Inflammation ovalbumin

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ovalbumin-loaded paramagnetic nano-triangles for enhanced dendritic cell stimulation,T_(1)-MR imaging,and antitumor immunity

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Journal of Materials Science & Technology 2023年第17期148卷 123-137页

作者： Yike Hou Zhe Tang Jabeen Farheen Madiha Saeed Lijia Luo Wenzi Ren Dandan Luo Asim Mushtaq Ruibo Zhao Jian Ge Zhangsen Yu Yao Li M.Zubair Iqbal Xiangdong Kong Institute of Smart Biomedical Materials School of Materials Science and EngineeringZhejiang Sci-Tech UniversityHangzhou 310018China State Key Laboratory of Drug Research&Center of Pharmaceutics Shanghai Institute of Materia MedicaChinese Academy of SciencesShanghai 201203China Cixi Institute of Biomedical Engineering CAS Key Laboratory of Magnetic Materials and DevicesKey Laboratory of Additive Manufacturing Materials of Zhejiang ProvinceNingbo Institute of Materials Technology and EngineeringChinese Academy of SciencesNingbo 315201China Department of Materials Imperial College LondonLondon SW72AZUK College of Life Sciences China Jiliang UniversityHangzhou 310018China Laboratory of Nanomedicine Medical Science Research CenterSchool of MedicineShaoxing UniversityShaoxing 312000China

Vaccine-based cancer immunotherapy has demonstrated a significant potential for cancer treatment in clinics.Although the efficiencies of vaccines are limited,they can be enhanced by a well-designed antigen delivery system that promotes sufficient antigen presentation of dendritic cells(DCs)for initiating high T cell immunity.Herein,antigen-loaded manganese oxide(Mn_(3)O_(4))triangular-shaped ultrasmall nanoparti-cles were prepared to stimulate DC-based immunotherapy under the guidance of T_(1)magnetic resonance imaging.The FDA-approved triblock copolymer Pluronic^(■)F-68 wasused not onlyto transferthe phase from hydrophobic to hydrophilic but also to enrich antigen loading and improve the biocompatibility of the prepared nanoparticles.ovalbumin(OVA),a model antigen,was adsorbed on the surface of polymer-coated nanoparticles through electrostatic interaction to form Mn_(3)O_(4)@PF68-OVA nanoparticle-antigen complexes to stimulate DC-based immunization and antigen-specific T cell immunity.The Mn_(3)O_(4)@PF68-OVA nanovaccine(NV)induces negligible toxicity effects against 4T1 and bone marrow-derived dendritic cells(BMDCs)by conventional methods supports the proliferation of intestine organoids,which are an innovative three-dimensional cytotoxicity evaluation system,thereby indicating their potential safety for in vivo cancer therapies.The designed paramagnetic nanovaccine possessed excellent OVA delivery to dendritic-regulated antigen-specific T cells in vitro by stimulating the maturation level of BMDCs.In ad-dition,Mn_(3)O_(4)@PF68-OVA NVs enhance immunity in vivo by increasing the T-cells and M1 macrophages,which suggests improved immunity.Excitingly,vaccination with Mn_(3)O_(4)@PF68-OVA offer complete pro-tection in the prophylactic group and significant tumor inhibition in the therapeutic group against B16-OVA tumor.In addition,the designed nanovaccine demonstrated high T_(1)-MR imaging in the tumor,fur-ther justifying enhanced tumor accumulation and capability to real-time monitor the treatment proce-dure.This

关键词： ovalbumin Nanovaccine Antigen delivery Magnetic resonance imaging Contrast agent Cancer immunotherapy

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Molecular dynamics simulations of ovalbumin adsorption at squalene/water interface

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Chinese Journal of Chemical Engineering 2022年第10期50卷 369-378页

作者： Qingxia Xiong Ying Ren Yufei Xia Guanghui Ma Reiji Noda Wei Ge State Key Laboratory of Multiphase Complex Systems Institute of Process EngineeringChinese Academy of SciencesBeijing 100190China Division of Environmental Engineering Science Graduate School of Science and TechnologyGunma University1-5-1 Tenjin-choKiryuGunma 376-8515Japan State Key Laboratory of Biochemical Engineering Institute of Process EngineeringChinese Academy of SciencesBeijing 100190China School of Chemical Engineering University of Chinese Academy of SciencesBeijing 100049China Innovation Academy for Green Manufacture Chinese Academy of SciencesBeijing 100190China

The adsorption of protein molecules to oil/water(O/W)interface is of critical importance for the product design in a wide range of technologies and industries such as biotechnology,food industry and pharmaceutical industry.In this work,with ovalbumin(OVA)as the model protein,the adsorption conformations at the O/W interface and the adsorption stability have been systematically studied via multiple simulation methods,including all-atom molecular dynamic(AAMD)simulations,coarse-grained molecular dynamic(CGMD)simulations and enhanced sampling methods.The computational results of AAMD and CGMD show that the hydrophobic tail of OVA tends to be folded under long time relaxation in aqueous phase,and multiple adsorption conformations can exist at the interface due to heterogeneous interactions raising from oil and water respectively.To further study the adsorption sites of the protein,the adsorption kinetics of OVA at the O/W interface is simulated using metadynamics method combined with CGMD simulations,and the result suggests the existence of multiple adsorption conformations of OVA at interface with the head-on conformation as the most stable one.In all,this work focuses on the adsorption behaviors of OVA at squalene/water interface,and provides a theoretical basis for further functionalization of the proteins in emulsion-based products and engineering.

关键词： Molecular dynamicssimulation Metadynamics Protein adsorption Structural stability ovalbumin

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MBD2 promotes Th2 differentiation in ovalbumin-induced CD4+T cells

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BIOCELL 2023年第11期47卷 2495-2502页

作者： QILU PAN YAN JIANG LINQIAO LI XIAOJING DU QIAN HAN FEIXIANG LING ROU LI SHUYUAN CHU LIN MAI JIANWEI HUANG LIBING MA Department of Respiratory and Critical Care Medicine Affiliated Hospital of Guilin Medical UniversityGuilin541001China Laboratory of Respiratory Disease Affiliated Hospital of Guilin Medical UniversityGuilin541001China

Introduction:Allergen-specific CD4+T cells play a central role in autoimmune disorders,allergies and asthma,with Th2-type immunity being the typical functional response of CD4+T cells.This study aimed to investigate the role of MBD2 in regulating Th2 cell differentiation.Methods:Splenic mononuclear cells were extracted from C57BL/6 mice,and CD4+T cells were isolated using magnetic beads and confirmed through flow cytometry.Lentivirus was employed to construct MBD2-silenced CD4+T cells.In vitro experiments were performed to treat splenogenic mononuclear cells and CD4+T cells with ovalbumin(OVA),and Th2 cell ratios and IL-4 levels were assessed using flow cytometry and ELISA.Results:The purity of the isolated CD4+T cells was 95.73%,confirming successful isolation of primary CD4+T cells.Compared to the control group,the Th2 cell ratio exhibited an increase in the Th2-induced group.Treatment with 5-Aza(concentrations,1-100μM)promoted Th2 cell differentiation and increased IL-4 levels.Notably,when combined with Th2 induction and 10μM 5-Aza treatment,silencing MBD2 further amplified Th2 cell ratios and elevated IL-4 levels in cell supernatants.Furthermore,OVA(concentration,200μg/mL)induced the differentiation of CD4+T cells into Th2 cells and increased IL-4 secretion.Interestingly,silencing MBD2 significantly increased the Th2 cell ratio and IL-4 levels in OVA-treated CD4+T cells.Conclusion:In summary,OVA promoted CD4+T cell differentiation into Th2 cells and enhanced IL-4 levels.MBD2 was identified as a mediator of Th2 cell differentiation in splenic-derived CD4+T cells,influenced by OVA or 5-Aza treatment.

关键词： 5-Aza MBD2 CD4+T cells Th2 cells ovalbumin

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Myricetin alleviates ovalbumin-induced allergic rhinitis in mice by regulating Th1/Th2 balance

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Asian Pacific Journal of Tropical Biomedicine 2023年第7期13卷 306-314页

作者： Ya-Nan Shi Jin-Zhu Su Juan Wang Jiang-Qiao Geng Department of Otolaryngology Sanya Women and Children's HospitalSanya572000China Department of Otolaryngology Hebei Children’s HospitalShijiazhuang050031China

Objective:To evaluate the effect of myricetin on ovalbumin(OVA)-induced allergic rhinitis in mice.Methods:Mice were sensitized and challenged using OVA(5%,500 mL)intraperitoneally and intranasally,respectively,on an alternative day for 14 days,followed by administration of myricetin(50,100,and 200 mg/kg)till day 21.Nasal symptoms,biochemical parameters,protein expressions,and histopathology were observed.Results:OVA-induced increased nasal symptoms including rubbing,sneezing,and discharge were significantly reduced by myricetin(100and 200 mg/kg)(P<0.05).Myricetin also protected against histamine challenge and attenuated elevated serum immunoglobulin E(IgE;total and OVA-specific),total IgG1,andβ-hexosaminidase levels,as well as leukotriene C4 and interleukins levels in nasal lavage fluid(P<0.05).Western blot analysis showed that myricetin significantly upregulated the protein expression of T-box expressed in T cells,while downregulating the protein expression of GATA binding protein 3,NF-κB,and IκB-α(P<0.05).Additionally,OVA-induced histopathological abberations in the nasal mucosa was markedly ameliorated by myricetin treatment(P<0.05).Conclusions:Myricetin exerts anti-allergic effects against OVAinduced allergic rhinitis via regulating Th1/Th2 balance.

关键词： Allergic rhinitis GATA3 Interleukins Myricetin Montelukast ovalbumin Th1/Th2 balance

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Effects of phosphodiesterase 4 inhibitor on cough response in guinea pigs sensitized and challenged with ovalbumin

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Chinese Medical Journal 2004年第11期117卷 1620-1624页

作者：吕寒静邱忠民魏为利余莉刘瑞麟张敏 Department of Respiratory Medicine Tongji Hospital Tongji University School of Medicine Shanghai 200065 China

BACKGROUND: There is currently considerable interest in the potential value of selective inhibitors of cyclic nucleotide phosphodiesterase 4 in the treatment of asthma. However, whether they influence eosinophilic airway inflammation-associated cough remains unclear. The objective of this study was to investigate the effects of selective phosphodiesterase 4 inhibitor SB207499 on cough response and airway inflammation in guinea pigs sensitized and challenged with ovalbumin. METHODS: Forty sensitized guinea pigs were randomly divided into four groups: control (n = 10), challenge (n = 10), SB207499 (n = 10) and aminophylline (n = 10), then challenged with aerosol of 1% ovalbumin or saline. Two hours later, animals were intraperitoneally injected with either saline, 25 mg/kg of SB207499 or aminophylline. At the 24th hour, the injection was repeated with 2.5 mg/kg and 25 mg/kg SB207499 or aminophylline, then cough response to inhaled capsaicin and airway responsiveness to methacholine inducing a 150% of the peak airway pressure to the baseline (PC150) was measured. Finally, total cell number and differentials in bronchoalveolar lavage fluid were analysed. RESULTS: The cough frequency per 3 minutes and PC150 in the challenge group were (22 +/- 4) times/3 minutes and (198 +/- 54) microg/ml, which were significantly different from (6 +/- 2) times/3 minutes and (691 +/- 81) microg/ml in the control group (P

关键词： 3',5'-Cyclic-Nucleotide Phosphodiesterase Animals Bronchial Hyperreactivity Bronchoalveolar Lavage Fluid Cough Cyclic AMP Cyclohexanecarboxylic Acids Dose-Response Relationship, Drug Guinea Pigs Male ovalbumin Phosphodiesterase Inhibitors

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Protective effect of Bifidobacterium infantis CGMCC313-2 on ovalbumin-induced airway asthma and b-lactoglobulininduced intestinal food allergy mouse models

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World Journal of Gastroenterology 2017年第12期23卷 2149-2158页

作者： Meng-Yun Liu Zhen-Yu Yang Wen-Kui Dai Jian-Qiong Huang Yin-Hu Li Juan Zhang Chuang-Zhao Qiu Chun Wei Qian Zhou Xin Sun Xin Feng Dong-Fang Li He-Ping Wang Yue-Jie Zheng Department of Respiratory Shenzhen Children's Hospital WeHealth Gene Co. Xijing Hospital The Fourth Military Medical University

AIM To determine whether oral administration of Bifidobacterium infantis CGMCC313-2(B. infantis CGMCC313-2) inhibits allergen-induced airway inflammation and food allergies in a mouse model.METHODS ovalbumin(OVA)-induced allergic asthma and b-lactoglobulin-induced food allergy mouse models were used in this study. Following oral administration of B. infantis CGMCC313-2 during or after allergen sensitization, histopathologic changes in the lung and intestine were evaluated by hematoxylin and eosin(HE) staining. In the allergic asthma mouse model, we evaluated the proportion of lung-infiltrating inflammatory cells. OVAspecific IgE and IgG1 levels in serum and cytokine levels in bronchoalveolar lavage fluid(BALF) were also assessed. In the food allergy mouse model, the levels of total Ig E and cytokines in serum were measured.RESULTS Oral administration of B. infantis CGMCC313-2 during or after allergen sensitization suppressed allergic inflammation in lung and intestinal tissues, while the proportion of infiltrating inflammatory cells was significantly decreased in the BALF of allergic asthma mice. Moreover, B. infantis CGMCC313-2 decreased the serum levels of total Ig E in food allergy mice, and reductions in IgE and IgG1 were also observed in OVA-induced allergic asthma mice. The expression of interleukin-4(IL-4) and IL-13 in both serum and BALF was suppressed following the administration of B. infantis CGMCC313-2, while an effect on serum IL-10 levels was not observed.CONCLUSION B. infantis CGMCC313-2 inhibits the secretion of allergen-induced IgE, IL-4 and IL-13, and attenuates allergic inflammation.

关键词： Bifidobacterium infantis Asthma Allergy ovalbumin b-lactoglobulin

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Cyclodextrin/chitosan nanoparticles for oral ovalbumin delivery: Preparation, characterization and intestinal mucosal immunity in mice

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Asian Journal of Pharmaceutical Sciences 2019年第2期14卷 193-203页

作者： Muye He Chen Zhong Huibing Hu Yu Jin Yanzuo Chen Kaiyan Lou Feng Gao Department of Pharmaceutics School of Pharmacy East China University of Science and Technology Shanghai Key Laboratory of New Drug Design East China University of Science and Technology Shanghai Key Laboratory of Functional Materials Chemistry East China University of Science and Technology

A novel oral protein delivery system with enhanced intestinal penetration and improved antigen stability based on chitosan(CS) nanoparticles and antigen-cyclodextrin(CD) inclusion complex was prepared by a precipitation/coacervation method. ovalbumin(OVA) as a model antigen was firstly encapsulated by cyclodextrin, either β-cyclodextrin( β-CD) or carboxymethyl-hydroxypropyl-β-cyclodextrin(CM-HP-β-CD) and formed OVA-CD inclusion complexes, which were then loaded to chitosan nanoparticles to form OVA loaded β-CD/CS or CM-HP-β-CD/CS nanoparticles with uniform particle size(836.3 and 779.2 nm, respectively) and improved OVA loading efficiency(27.6% and 20.4%, respectively). In vitro drug release studies mimicking oral delivery condition of OVA loaded CD/CS nanoparticles showed low initial releases at p H 1.2 for 2 h less than 3.0% and a delayed release which was below to 30% at p H 6.8 for further 72 h. More importantly, after oral administration of OVA loaded β-CD/CS nanoparticles to Balb/c mice, OVA-specific sIgA levels in jejunum of OVA loaded β-CD/CS nanoparticles were 3.6-fold and 1.9-fold higher than that of OVA solution and OVA loaded chitosan nanoparticles, respectively. In vivo evaluation results showed that OVA loaded CD/CS nanoparticles could enhance its efficacy for inducing intestinal mucosal immune response. In conclusion, our data suggested that CD/CS nanoparticles could serve as a promising antigen-delivery system for oral vaccination.

关键词： β-cyclodextrin Chitosan nanoparticles ovalbumin Oral protein delivery Intestinal mucosal immunity

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