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An In vitro Model to Study Heterogeneity of Human Macrophage Differentiation and Polarization
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Atypical Activin A and IL-10 Production Impairs Human CD16+ Monocyte Differentiation into Anti-Inflammatory

Érika González-Domínguez1, Ángeles Domínguez-Soto2, Concha Nieto2

  • 1Departamento de Biomedicina Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, 07360 Mexico City, Mexico;

Journal of Immunology (Baltimore, Md. : 1950)
|January 6, 2016
PubMed

Insights

Human monocytes CD14(++)CD16(-) differentiate into anti-inflammatory macrophages, while CD16(+) monocytes become proinflammatory macrophages unless treated with IL-10. This reveals distinct macrophage differentiation pathways.

Area of Science:

  • Immunology
  • Cell Biology
  • Molecular Biology

Background:

  • Human blood monocytes consist of CD14(++)CD16(-) (85%) and CD14(+/lo)CD16(+) (15%) subsets, presumed to represent distinct differentiation stages.
  • The specific differentiation fates of these monocyte subsets into macrophages (Mϕ) and their inflammatory polarization remain incompletely understood.

Purpose of the Study:

  • To investigate the differentiation potential and inflammatory priming of CD14(++)CD16(-) and CD16(+) monocyte subsets.
  • To elucidate the mechanisms underlying the distinct Mϕ polarization induced by M-CSF or GM-CSF in these subsets.

Main Methods:

  • Isolation and culture of CD14(++)CD16(-) and CD16(+) monocyte subsets.
  • Differentiation induction using M-CSF or GM-CSF, generating M14, M16, GM14, and GM16.
  • Phenotypic and functional analysis of differentiated Mϕs, including transcriptomic profiling and cytokine measurements.
  • Intervention studies using activin A receptor blockade and IL-10 supplementation.

Main Results:

  • GM-CSF-treated monocytes (GM14, GM16) were phenotypically and functionally similar.
  • M-CSF-treated CD16(+) monocytes (M16) exhibited a more proinflammatory profile than M-CSF-treated CD14(++)CD16(-) monocytes (M14).
  • M16 cells showed underrepresentation of M-CSF-driven Mϕ differentiation genes and higher activin A secretion, with lower IL-10 production compared to M14.
  • Blocking activin A or adding IL-10 to M16 cultures shifted their phenotype towards anti-inflammatory Mϕ markers.

Conclusions:

  • M-CSF drives CD14(++)CD16(-) monocyte differentiation into anti-inflammatory Mϕs autonomously.
  • M-CSF-treated CD16(+) monocytes differentiate into Mϕs with a proinflammatory skew due to high activin A and low IL-10 levels.
  • Providing additional anti-inflammatory stimuli like IL-10 can modulate the M16 differentiation towards an anti-inflammatory phenotype.

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