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Immune-triggering effect of the foodborne parasite Kudoa septempunctata through the C-type lectin Mincle in HT29 cells

BMB Reports 2020년 53권 9호 p.478 ~ 483
신지현, 양정표, 서승환, 김상균, 김은민, 함도원, 신은희,
소속 상세정보
신지현 ( Shin Ji-Hun ) - Seoul National University College of Medicine Department of Parasitology and Tropical Medicine
양정표 ( Yang Jung-Pyo ) - Seoul National University College of Medicine Department of Parasitology and Tropical Medicine
서승환 ( Seo Seung-Hwan ) - Seoul National University College of Medicine Department of Parasitology and Tropical Medicine
김상균 ( Kim Sang-Gyun ) - Seoul National University College of Medicine Department of Parasitology and Tropical Medicine
김은민 ( Kim Eun-Min ) - Yonsei University College of Medicine Department of Environmental Medical Biology
함도원 ( Ham Do-Won ) - Seoul National University College of Medicine Department of Parasitology and Tropical Medicine
신은희 ( Shin Eun-Hee ) - Seoul National University College of Medicine Department of Parasitology and Tropical Medicine

Abstract


Kudoa septempunctata is a myxozoan parasite that causes food poisoning in individuals consuming olive flounder. The present study aimed to investigate the currently insufficiently elucidated early molecular mechanisms of inflammatory responses in the intestine owing to parasite ingestion. After Kudoa spores were isolated from olive flounder, HT29 cells were exposed to spores identified to be alive using SYTO-9 and propidium iodide staining or to antigens of Kudoa spores (KsAg). IL-1β, IL-8, TNF-α and NFKB1 expression and NF-κB activation were assessed using real-time PCR, cytokine array and western blotting. The immunofluorescence of FITC-conjugated lectins, results of ligand binding assays using Mincle-Fc and IgG-Fc, CLEC4E expressions in response to KsAg stimulation, and Mincle-dependent NF-κB activation were assessed to clarify the early immunetriggering mechanism. Inflammatory cytokines (IL-1α, GM-CSF and TNF-α), chemokines (IL-8, CCL2, CCL5 and CXCL1) and NF-κB activation (pNF-κB/NF-κB) in HT29 cells increased following stimulation by KsAg. The immunofluorescence results of spores and lectins (concanavalin A and wheat germ agglutinin) suggested the importance of Mincle in molecular recognition between Kudoa spores and intestinal cells. Practically, data for Mincle-Fc and KsAg binding affinity, CLEC4E mRNA expression, Mincle immunofluorescence staining and hMincledependent NF-κB activation demonstrated the involvement of Mincle in the early immune-triggering mechanism. The present study newly elucidated that the molecular recognition and immune-triggering mechanism of K. septempunctata are associated with Mincle on human intestinal epithelial cells.

키워드

Cell adhesion; C-type lectin Mincle; Innate immunity; Myxozoan parasite; Signal transduction

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