{"title":"Role of Raf Kinase Inhibitor Protein in Modulating Echinococcal Cyst Fluid-induced Degranulation in Bone Marrow-Derived Mast Cells.","authors":"Shan-Shan Li, Xue-Li Pu, Jing-Ru Zhou, Chun-Sheng Wang, Jia-Ling Wang, Xilizati Kulaixi, Jian-Rong Ye","doi":"10.1159/000545176","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>This study aimed to investigate the role of Raf kinase inhibitor protein (RKIP) in degranulation induced by echinococcal cyst fluid (EgCF) in bone marrow-derived mast cells (BMMCs).</p><p><strong>Methods: </strong>Primary BMMCs were isolated and cultured from the femurs and tibias of RKIP gene knockout (KO) and wild-type (WT) C57BL/6 mice. EgCF-induced degranulation models were established for both groups. Samples of cells and supernatant were collected for analysis. Surface expression levels of CD117 and Fc-epsilon Receptor Ⅰ α (FcεRIα) were assessed. Supernatant concentrations of β-hexosaminidase, IL-4, IL-6, and tumor necrosis factor (TNF-α) were measured. Cellular mRNA levels of IL-4, IL-6, and TNF-α were quantified, and changes in RKIP protein expression during degranulation in WT BMMCs were monitored.</p><p><strong>Results: </strong>After 4 weeks of induction culture, the double-positive rates for CD117 and FcεRIα exceeded 98% in both KO and WT BMMCs. Following sensitization, BMMCs from the KO group demonstrated significantly higher degranulation rates compared to the WT group (p < 0.05). In WT BMMCs, surface RKIP protein expression progressively decreased at 1 hour, 2 hours, and 3 hours post-sensitization, corresponding with degranulation progression (p < 0.05). The KO group exhibited elevated release of BMMC-related cytokines, including IL-4, IL-6, and TNF-α, compared to the WT group after sensitization (p < 0.05). Similarly, transcription levels of cytokines IL-4, IL-6, and TNF-α were higher in the KO group than in the WT group following sensitization (p < 0.05).</p><p><strong>Conclusion: </strong>RKIP gene knockout resulted in an increased EgCF-induced release of cytokines and bioactive substances by BMMCs, indicating that RKIP may suppress EgCF-induced BMMC degranulation. These findings suggest that RKIP could serve as a potential therapeutic target for managing allergic reactions associated with cystic echinococcosis.</p>","PeriodicalId":13652,"journal":{"name":"International Archives of Allergy and Immunology","volume":" ","pages":"1-12"},"PeriodicalIF":2.5000,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"International Archives of Allergy and Immunology","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1159/000545176","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"ALLERGY","Score":null,"Total":0}
引用次数: 0
Abstract
Objective: This study aimed to investigate the role of Raf kinase inhibitor protein (RKIP) in degranulation induced by echinococcal cyst fluid (EgCF) in bone marrow-derived mast cells (BMMCs).
Methods: Primary BMMCs were isolated and cultured from the femurs and tibias of RKIP gene knockout (KO) and wild-type (WT) C57BL/6 mice. EgCF-induced degranulation models were established for both groups. Samples of cells and supernatant were collected for analysis. Surface expression levels of CD117 and Fc-epsilon Receptor Ⅰ α (FcεRIα) were assessed. Supernatant concentrations of β-hexosaminidase, IL-4, IL-6, and tumor necrosis factor (TNF-α) were measured. Cellular mRNA levels of IL-4, IL-6, and TNF-α were quantified, and changes in RKIP protein expression during degranulation in WT BMMCs were monitored.
Results: After 4 weeks of induction culture, the double-positive rates for CD117 and FcεRIα exceeded 98% in both KO and WT BMMCs. Following sensitization, BMMCs from the KO group demonstrated significantly higher degranulation rates compared to the WT group (p < 0.05). In WT BMMCs, surface RKIP protein expression progressively decreased at 1 hour, 2 hours, and 3 hours post-sensitization, corresponding with degranulation progression (p < 0.05). The KO group exhibited elevated release of BMMC-related cytokines, including IL-4, IL-6, and TNF-α, compared to the WT group after sensitization (p < 0.05). Similarly, transcription levels of cytokines IL-4, IL-6, and TNF-α were higher in the KO group than in the WT group following sensitization (p < 0.05).
Conclusion: RKIP gene knockout resulted in an increased EgCF-induced release of cytokines and bioactive substances by BMMCs, indicating that RKIP may suppress EgCF-induced BMMC degranulation. These findings suggest that RKIP could serve as a potential therapeutic target for managing allergic reactions associated with cystic echinococcosis.
期刊介绍:
''International Archives of Allergy and Immunology'' provides a forum for basic and clinical research in modern molecular and cellular allergology and immunology. Appearing monthly, the journal publishes original work in the fields of allergy, immunopathology, immunogenetics, immunopharmacology, immunoendocrinology, tumor immunology, mucosal immunity, transplantation and immunology of infectious and connective tissue diseases.