Characterization of phenotype markers and neuronotoxic potential of polarised primary microglia In vitro

Chhor, V, Le Charpentier, T, Lebon, S, Ore, M, Celador, I, Josserand, J, Degos, V, Jacotot, E, Hagberg, H, Savman, K, Mallard, C, Gressens, P and Fleiss, B 2013, 'Characterization of phenotype markers and neuronotoxic potential of polarised primary microglia In vitro', Brain, Behavior, and Immunity, vol. 32, pp. 70-85.

Document type: Journal Article
Collection: Journal Articles

Title Characterization of phenotype markers and neuronotoxic potential of polarised primary microglia In vitro
Author(s) Chhor, V
Le Charpentier, T
Lebon, S
Ore, M
Celador, I
Josserand, J
Degos, V
Jacotot, E
Hagberg, H
Savman, K
Mallard, C
Gressens, P
Fleiss, B
Year 2013
Journal name Brain, Behavior, and Immunity
Volume number 32
Start page 70
End page 85
Total pages 16
Publisher Academic Press
Abstract Microglia mediate multiple facets of neuroinflammation, including cytotoxicity, repair, regeneration, and immunosuppression due to their ability to acquire diverse activation states, or phenotypes. Modulation of microglial phenotype is an appealing neurotherapeutic strategy but a comprehensive study of classical and more novel microglial phenotypic markers in vitro is lacking. The aim of this study was to outline the temporal expression of a battery of phenotype markers from polarised microglia to generate an in vitro tool for screening the immunomodulatory potential of novel compounds. We characterised expression of thirty-one macrophage/microglial phenotype markers in primary microglia over time (4, 12, 36, and 72 h), using RT-qPCR or multiplex protein assay. Firstly, we selected Interleukin-4 (IL-4) and lipopolysaccharide (LPS) as the strongest M1-M2 polarising stimuli, from six stimuli tested. At each time point, markers useful to identify that microglia were M1 included iNOS, Cox-2 and IL-6 and a loss of M2a markers. Markers useful for quantifying M2b-immunomodulatory microglia included, increased IL-1RA and SOCS3 and for M2a-repair and regeneration, included increased arginase-1, and a loss of the M1 and M2b markers were discriminatory. Additional markers were regulated at fewer time points, but are still likely important to monitor when assessing the immunomodulatory potential of novel therapies. Further, to facilitate identification of how novel immunomodulatory treatments alter the functional affects of microglia, we characterised how the soluble products from polarised microglia affected the type and rate of neuronal death; M1/2b induced increasing and M2a-induced decreasing neuronal loss. We also assessed any effects of prior activation state, to provide a way to identify how a novel compound may alter phenotype depending on the stage of injury/insult progression. We identified generally that a prior M1/2b reduced the ability of microglia to switch to M2a
Subject Innate Immunity
Neurology and Neuromuscular Diseases
Keyword(s) Drug-screening
Neuronal cell death
DOI - identifier 10.1016/j.bbi.2013.02.005
Copyright notice © 2013 Elsevier Inc.
ISSN 0889-1591
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