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

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-1, IL-8, TNF- and NFKB1 expression and NF-B 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-B activation were assessed to clarify the early immunetriggering mechanism. Inflammatory cytokines (IL-1, GM-CSF and TNF-), chemokines (IL-8, CCL2, CCL5 and CXCL1) and NF-B activation (pNF-B/NF-B) 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-B 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. [BMB Reports 2020; 53(9): 478-483]

Kudoa septempunctata Spores Cause Acute Gastroenteric Symptoms in Mouse and Musk Shrew Models as Evidenced In Vitro in Human Colon Cells

“…The sporoplasm released from K . septempunctata spores contributes to gastrointestinal symptoms and occasionally results in concurrent inflammatory symptoms [ 6 , 16 , 24 , 25 ].…”

“…The habit of eating raw fish therefore poses a high risk to the integrity of the intestinal mucosa [5,7,23]. The sporoplasm released from K. septempunctata spores contributes to gastrointestinal symptoms and occasionally results in concurrent inflammatory symptoms [6,16,24,25].…”

“…Invasion results in the rapid decrease in transepithelial electrical resistance (TEER) [15]. Another previous study reported the immune-promoting effects of K. septempunctata in HT29 cells [16]. Conversely, another study reported that K. septempunctata spores did not cause inflammation in Caco-2 cells [13].…”

“…Another previous study reported the immune-promoting effects of K . septempunctata in HT29 cells [ 16 ]. Conversely, another study reported that K .…”

Macrophage‐inducible C‐type lectin (MINCLE): In silico characterization and its in vitro effects on head–kidney leukocytes from the fishLutjanus peru

“…The macrophage‐inducible C‐type lectin (MINCLE) receptor in mammals recognizes 6‐trehalose, glycolipids, mannose, and β‐glucans (Matsumoto et al, 1999; Ishikawa et al, 2009; Ishikawa et al, 2013, Hattori et al, 2014; Hara et al, 2007). Downstream signalling pathways, mediated by the immunoreceptor tyrosine‐based activation motif, activate pro‐inflammatory cytokine production (Wells et al, 2008; Shin et al, 2020; Zhang et al, 2018). In fish, Mincle expression followed by NF‐κβ has been observed with the subsequent production of IL‐1β, IL‐8 and TNF‐α (Brietzke et al, 2016).…”