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Antimicrobial immunity involves immuno-metabolic regulation induced by CpG-ODN in broiler chickens

2022 CPRF,Health & Immunity
Oligodeoxynucleotides containing Cytosine phosphodiester Guanine motifs (CpG-ODNs), synthetic short segments of DNA represent a type of pathogen-associated molecular pattern (PAMP) that can stimulate immune cells and promote antimicrobial immunity. Activated immune cells undergo profound metabolic changes to meet cellular signaling process and biosynthesis demands. We hypothesize that the stimulation of immune cells by CpG-ODNs promotes profound metabolic programming to facilitate the antimicrobial activities of immune cells. We administered CpG-ODN (two doses, at day 1 and day 3) or saline into broiler chicks via the intramuscular route and longitudinally evaluated immunometabolic responses in the blood leukocytes. Peripheral blood mononuclear cell (PBMC) layer was isolated and processed to measure cellular mitochondrial activity [oxygen consumption rate (OCR)] and glycolysis rate [extracellular acidification rate (ECAR)] in real-time using a Seahorse XFp Extracellular Flux Analyzer (Seahorse Bioscienice). To identify whether T cells or B cells are responsible for CpG-ODN mediated protection, PBMC layer was incubated with Pokeweed mitogen (PWM) and Phytohaemagglutinin (PHA-P) separately and OCR was quantified. We observed a significantly enhanced cellular metabolic capacity (mitochondrial respiration and glycolysis) in CpG-ODN injected group than saline control. CpG-ODN injected group showed higher, statistically significant (p<0.001) mitochondrial respiration than the saline group at 24 hours, 48 hours (the highest), day 7, and day 14-time points. Birds that received CpG-ODN had increased glycolytic capacity (only at 48 hours’ time-point) in CpG-ODN group compared to the control group. These data indicate a substantial elevation of immune cell metabolism, especially in mitochondrial respiration. Further, PWM (mitogenic for both T and B cells) stimulated cells showed higher (p<0.05) OCR than PHA-P (mitogenic only for T cells) stimulated cells in the CpG-ODN group. These results indicate increased activity of B cells in CpG-ODN injected birds. Our study demonstrated that CpG-ODN associated antimicrobial activity increased energy demand and metabolic activities of immune cells in broiler chickens. These information will help to develop a potent alternative to antibiotics for the poultry industry.
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PhD Student

University of Saskatchewan

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