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  • C-type lectin 17A and macrophage-expressed receptor genes are magnified by fungal β-glucan after Vibrio parahaemolyticus infection in Totoaba macdonaldi cells.

C-type lectin 17A and macrophage-expressed receptor genes are magnified by fungal β-glucan after Vibrio parahaemolyticus infection in Totoaba macdonaldi cells.

Immunobiology (2018-12-01)
Carlos Angulo, Veronica Sanchez, Karen Delgado, Martha Reyes-Becerril
ABSTRACT

C-type lectins are a principal carbohydrate recognition mechanism as glucans on cell surfaces. This study identified and investigated molecular characterization and immune roles of a novel c-type lectin 17A from Totoaba macdonaldi (TmCLEC17A), which were described in head-kidney leukocytes after immunostimulation with fungal β-glucan 197A and Vibrio parahaemolyticus infection. This nucleotide sequence from totoaba was acquired using NGS and bioinformatics tools. Its full-length cDNA sequence consisted of 1128 bp (including the stop codon) and an open reading frame (ORF) of 771 bp encoding a 256 amino acid protein, 5´-UTR of 48 bp and 3´-UTR of 309 bp. The TmCLEC17A protein revealed a C-terminal-C-type lectin (CTL, also named carbohydrate-recognition domain, CRD), a N-terminal trans-membrane domain and a coiled coil motif, showing the highest similarity (80%) and identity (96%) with Larimichthys crocea. Fungal β-glucan 197A plus V. parahaemolyticus enhanced transcriptions of CLEC17A and TLR2 significantly besides the macrophage receptors, such as macrophage mannose receptor 1 and macrophage colony-stimulating factor 1 receptor 2. In addition, natural resistance-associated macrophage protein 2 was significantly up-regulated in leukocytes challenged with live V. parahaemolyticus. Overall, these results indicated that CLEC17A might be implicated in T. macdonaldi innate immunity as a pattern recognition receptor; fungal β-glucan 197A could stimulate cellular immune mechanisms in head-kidney leukocytes; and it could be used as potential immunostimulant in fish aquaculture.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Histopaque®-1077, sterile-filtered, density: 1.077 g/mL
Sigma-Aldrich
Zymosan A from Saccharomyces cerevisiae, for inducing inflamatory response