In vitro Phenotype Induction of Circulating Monocytes: CD16 and CD163 Analysis.
classical
intermediate
monocytes
non-classical
phenotype
stimulation
Journal
Journal of inflammation research
ISSN: 1178-7031
Titre abrégé: J Inflamm Res
Pays: New Zealand
ID NLM: 101512684
Informations de publication
Date de publication:
2021
2021
Historique:
received:
24
11
2020
accepted:
07
01
2021
entrez:
3
2
2021
pubmed:
4
2
2021
medline:
4
2
2021
Statut:
epublish
Résumé
CD14 (monocyte differentiation antigen, LPS binding protein - endotoxin receptor) and CD16 (FcγRIII, Low-affinity receptor for IgG) define three subpopulations of circulating monocytes with different inflammatory and phagocytic capabilities. Contradictory reports exist regarding both in vivo monocyte phenotype-disease association and response of these circulating monocytes to in vitro stimulation. We analyzed phenotypic changes in circulating monocytes when stimulated with LPS (pro-inflammatory stimulus) and IL-4 (alternative inflammatory stimulus). Mononuclear cells from nine healthy donors were extracted and studied for surface and intracellular markers using flow cytometry. PBMC were extracted using Ficoll technic and immediately analyzed using flow cytometry. Pro-inflammatory interleukin IL-1β and IL-6 were measured by intracellular cytometry. Mononuclear cells were stimulated using LPS and IL-4 as previously described. Changes against non-stimulated populations were statistically analyzed. Compared to non-stimulated and IL-4 stimulated monocytes, LPS-stimulated cells display a singular pattern of markers, with higher levels of intracellular IL-1β and IL-6 directly correlating with CD14+CD163- cell frequency and diminishing membrane CD163 fluorescence. CD14+CD16- classical monocytes show greater percentage of CD163- cells upon LPS stimulation. CD86 levels on monocytes' surface did not change with LPS or IL-4 stimulation. We showed that CD14+CD16- classical monocytes display higher sensitivity to LPS stimulation, with more IL-1β and IL-6 levels than intermediate and non-classical monocytes. This subset also diminishes its CD163 levels on the membrane after LPS stimulation with a contemporary raise in CD163- cells, suggesting that classical monocytes preferentially acquire CD163- defined M1 characteristics upon in vitro LPS stimulation. Intermediate and non-classical monocytes respond with lower levels of interleukins and display surface proteins in an M2-type profile (CD163+).
Identifiants
pubmed: 33531825
doi: 10.2147/JIR.S292513
pii: 292513
pmc: PMC7847382
doi:
Types de publication
Journal Article
Langues
eng
Pagination
191-198Informations de copyright
© 2021 Karsulovic et al.
Déclaration de conflit d'intérêts
The authors report no conflicts of interest for this work.
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