TLR-Mediated Cytokine Gene Expression in Chicken Peripheral Blood Mononuclear Cells As a Measure to Characterize Immunobiotics

Overview

Journal Genes (Basel)

Publisher MDPI

Date 2021 Feb 12

PMID 33572768

Citations 9

Authors

Anna Slawinska

Aleksandra Dunislawska

Arkadiusz Plowiec

Jose Goncalves

Maria Siwek

Affiliations

Soon will be listed here.

Abstract

Immunobiotics are probiotics that promote intestinal health by modulating immune responses. Immunobiotics are recognized by Toll-like receptors (TLRs) and activate cytokine gene expression. This study aimed to characterize cytokine gene expression in the chicken peripheral blood mononuclear cells (PBMC) stimulated with purified TLR ligands and live probiotics. PBMC were isolated from the whole blood. PBMC were stimulated with: lipopolysaccharide (LPS), CpG ODN, Pam3CSK4, Zymosan, galactooligosaccharides (GOS), subsp. (), and at 42.5 °C and 5% CO for 3 h, 6 h, and 9 h. After each time-point, PBMC were harvested for RNA isolation. Relative gene expression was analyzed with RT-qPCR for cytokine genes (, , , , , , , , and ) and reference genes ( and ). Genes were clustered into pro-inflammatory genes, Th1/Th2 genes, and Th1-regulators. The gene expression differed between treatments in , , , , and ( < 0.001). The genes , , and had the highest fold change of mRNA expression at 3 h in response to TLR ligands. up-regulated the pro-inflammatory genes at the 6 h time-point. did not activate the anti-inflammatory gene, but activated at 6 h. Hereby, was proven to exert immunostimulatory properties in PBMC.

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