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Clin Vaccine Immunol. 2016 Dec 5;23(12):926-933. Print 2016 Dec.

In Vitro Experimental Model of Trained Innate Immunity in Human Primary Monocytes.

Author information

1
Department of Internal Medicine, Radboud University Medical Center, Nijmegen, The Netherlands.
2
Research Center for Vitamins and Vaccines, Bandim Health Project, Statens Serum Institut, Copenhagen, Denmark.
3
Odense Patient Data Explorative Network, University of Southern Denmark/Odense University Hospital, Copenhagen, Denmark.
4
Department of Internal Medicine, Radboud University Medical Center, Nijmegen, The Netherlands mihai.netea@radboudumc.nl.

Abstract

Innate immune memory, or trained immunity, has recently been described to be an important property of cells of the innate immune system. Due to the increased interest in this important new field of immunological investigation, we sought to determine the optimal conditions for an in vitro experimental protocol of monocyte training using three of the most commonly used training stimuli from the literature: β-glucan, the bacillus Calmette-Guérin (BCG) vaccine, and oxidized low-density lipoprotein (oxLDL). We investigated and optimized a protocol of monocyte trained immunity induced by an initial training period with β-glucan, BCG, or oxLDL, followed by washing and resting of the cells and, thereafter, restimulation with secondary bacterial stimuli. The training and resting time intervals were varied to identify the optimal setting for the long-term induction of trained immunity. Trained immunity was assessed in terms of the secondary cytokine response, the production of reactive oxygen species, cell morphology, and induction of glycolysis. Monocytes primed with β-glucan, BCG, and oxLDL showed increased pro- and anti-inflammatory cytokine responses upon restimulation with nonrelated stimuli. Also, all three stimuli induced a switch to glycolysis (the Warburg effect). These effects were most pronounced when the training interval was 24 h and the resting time interval was 6 days. Training with BCG and oxLDL also led to the increased production of reactive oxygen species, whereas training with β-glucan led to the decreased production of reactive oxygen species. We describe the optimal conditions for an in vitro experimental model with human primary monocytes for study of the induction of trained innate immunity by microbial and metabolic stimuli.

PMID:
27733422
PMCID:
PMC5139603
DOI:
10.1128/CVI.00349-16
[Indexed for MEDLINE]
Free PMC Article

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