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Published on: November 15, 2024
Phenotypic and Functional Alterations in Peripheral Blood Mononuclear Cell-Derived Microglia in a Primate Model of
Hami Hemati1, Madison B Blanton2, Heather E True2
1Microbiology, Immunology and Molecular Genetics, College of Medicine, University of Kentucky, Lexington, KY, United States.
Abstract:
Alcohol-induced dysregulation of microglial activity is associated with neuroinflammation, cognitive decline, heightened risk for neurodegenerative diseases, alcohol dependence, and escalation of alcohol drinking. Given the challenge of longitudinally sampling primary microglia, we optimized an in vitro method to differentiate peripheral blood mononuclear cells (PBMC) from non-human primates (NHP) into microglia-like cells (induced-microglia; iMGL). The iMGLs displayed transcriptional profiles distinct from those of monocyte progenitors and closely resembling those of primary microglia. Notably, morphological features showed that differentiated iMGLs derived from NHPs with chronic alcohol consumption (CAC) possessed a more mature-like microglial morphology. Additionally, dysregulation in key inflammatory and regulatory markers alongside increased baseline phagocytic activity was observed in CAC-derived IMGLs in the resting state. Phenotypic and functional assessments following LPS stimulation indicated the presence of an immune-tolerant phenotype and enrichment of a CD86+ hyper-inflammatory subpopulation in iMGLs derived from ethanol-exposed animals. Collectively, these findings demonstrate that in vitro differentiation of PBMC offers a minimally invasive approach to studying the impact of CAC on microglial function revealing that CAC reshapes both functional and transcriptional profiles of microglia.
Insights
Chronic alcohol consumption alters microglia function and gene expression. This study developed an in vitro method using peripheral blood mononuclear cells to create microglia-like cells, revealing alcohol
Area of Science:
- Neuroscience
- Immunology
- Toxicology
Background:
- Alcohol-induced microglial dysregulation contributes to neuroinflammation and cognitive decline.
- Studying microglia longitudinally is challenging due to difficulties in accessing primary cells.
Purpose of the Study:
- To optimize an in vitro method for differentiating peripheral blood mononuclear cells (PBMC) into microglia-like cells (iMGL) from non-human primates (NHP).
- To investigate the impact of chronic alcohol consumption (CAC) on microglial function and phenotype using this novel method.
Main Methods:
- Differentiated PBMCs from NHP into iMGLs.
- Analyzed transcriptional profiles, morphology, and inflammatory markers of iMGLs.
- Assessed iMGL function following lipopolysaccharide (LPS) stimulation.
Main Results:
- iMGLs exhibited transcriptional profiles similar to primary microglia and distinct from monocyte progenitors.
- iMGLs from NHP with CAC showed mature morphology, increased baseline phagocytic activity, and dysregulated inflammatory markers.
- Ethanol-exposed iMGLs displayed an immune-tolerant phenotype and a hyper-inflammatory CD86+ subpopulation post-LPS stimulation.
Conclusions:
- In vitro differentiation of PBMC provides a minimally invasive approach to study microglial responses to chronic alcohol exposure.
- Chronic alcohol consumption reshapes both functional and transcriptional profiles of microglia, impacting neuroinflammation and immune response.
