Cited by 120Neuroinflammation and Neurodegeneration MechanismsImmune cells in cancerInflammasome and immune disorders

The DOI matches its Crossref record; 110 of the 115 references it cites were found in a registry or an index.

Loss of IL-10 Promotes Differentiation of Microglia to a M1 Phenotype

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Abstract

Microglia represent the primary resident immune cells of the central nervous system (CNS) and modulate local immune responses. Depending on their physiological functions, microglia can be classified into pro- (M1) and anti-inflammatory (M2) phenotype. Interleukin (IL)-10 is an important modulator of neuronal homeostasis, with anti-inflammatory and neuroprotective functions, and can be released by microglia. Here, we investigated how IL-10 deficiency affected the M1/2 polarization of primary microglia upon lipopolysaccharide (LPS) stimulation in vitro. Microglia phenotypes were analyzed via flow cytometry. Cytokine and chemokine secretion were examined by ELISA and bead-based multiplex LEGENDplex™. Our results showed that genetic depletion of IL-10 led to elevated M1 like phenotype (CD86+CD206-) under pro-inflammatory conditions associated with increased frequency of IL-6+, TNF-α+ cells and enhanced release of several pro-inflammatory chemokines. Absence of IL-10 led to an attenuated M2 like phenotype (CD86-CD206+) and a reduced secretion of TGF-β1 upon LPS stimulation. In conclusion, IL-10 deficiency may promote the polarisation of microglia into M1-prone phenotype under pro-inflammatory conditions.

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Crossref · 10.3389/fncel.2019.00430

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Figures

FIGURE 1 Fluorescence microscopy of Iba1+ WT and IL-10 KO microglia. (A) WT (control), (B) WT (LPS); slim cell bodies, (C) IL-10 KO (control), (D) IL-10 KO (LPS), round, amoeboid cell shape. Representative figures of four independent experiments (n = 4); overlay Iba1/green, nuclei/blue, 200-fold magnification.
FIGURE 1 Fluorescence microscopy of Iba1+ WT and IL-10 KO microglia. (A) WT (control), (B) WT (LPS); slim cell bodies, (C) IL-10 KO (control), (D) IL-10 KO (LPS), round, amoeboid cell shape. Representative figures of four independent experiments (n = 4); overlay Iba1/green, nuclei/blue, 200-fold magnification.CC BY · PubMed Central
FIGURE 2 Cytokine pattern of WT (black bars) or IL-10 KO (gray bars) microglia of control group or after LPS treatment. ELISA data in pg/ml, (A) IL-6 (N = 11), (B) TNF-α (N = 11), (C) IL-10 (N = 10), and (D) TGF-β1 (N = 8), Mean ± SEM. Flow cytometric data in % CD11b+ cells, (E) IL-6, (F) TNF-α, (G) IL-10, (H) TGF-β1 (N = 5), mean ± SEM. One-way ANOVA, ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ****p < 0.0001.
FIGURE 2 Cytokine pattern of WT (black bars) or IL-10 KO (gray bars) microglia of control group or after LPS treatment. ELISA data in pg/ml, (A) IL-6 (N = 11), (B) TNF-α (N = 11), (C) IL-10 (N = 10), and (D) TGF-β1 (N = 8), Mean ± SEM. Flow cytometric data in % CD11b+ cells, (E) IL-6, (F) TNF-α, (G) IL-10, (H) TGF-β1 (N = 5), mean ± SEM. One-way ANOVA, ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ****p < 0.0001.CC BY · PubMed Central
FIGURE 3 Chemokine release (Δ: LPS minus control) of WT (black bars) and IL-10 KO (gray bars) microglia under LPS treatment. LEGENDplexTM data in pg/ml, (A) CCL2, (B) CCL3, (C) CCL4, (D) CCL5, (E) CCL11, (F) CCL17, (G) CCL20, (H) CCL22, (I) CXCL1, (J) CXCL5, (K) CXCL9, (L) CXCL10, and (M) CXCL13 (N = 8), mean ± SEM. Unpaired t-test, ∗p < 0.05.
FIGURE 3 Chemokine release (Δ: LPS minus control) of WT (black bars) and IL-10 KO (gray bars) microglia under LPS treatment. LEGENDplexTM data in pg/ml, (A) CCL2, (B) CCL3, (C) CCL4, (D) CCL5, (E) CCL11, (F) CCL17, (G) CCL20, (H) CCL22, (I) CXCL1, (J) CXCL5, (K) CXCL9, (L) CXCL10, and (M) CXCL13 (N = 8), mean ± SEM. Unpaired t-test, ∗p < 0.05.CC BY · PubMed Central
FIGURE 4 Flow cytometry of WT (black bars) and IL-10 KO (gray bars) microglia of control group or after LPS treatment. Data in% of CD11b+ cells, (A) CD11b, (B) CCR2, (C) CX3CR1, (D) M1 CD86+ CD206–, (E) M2 CD86-CD206+, (F) M1/2 CD86+ CD206+ (N = 3), mean ± SEM. One-way ANOVA, ∗p < 0.05; ∗∗∗p < 0.001.
FIGURE 4 Flow cytometry of WT (black bars) and IL-10 KO (gray bars) microglia of control group or after LPS treatment. Data in% of CD11b+ cells, (A) CD11b, (B) CCR2, (C) CX3CR1, (D) M1 CD86+ CD206–, (E) M2 CD86-CD206+, (F) M1/2 CD86+ CD206+ (N = 3), mean ± SEM. One-way ANOVA, ∗p < 0.05; ∗∗∗p < 0.001.CC BY · PubMed Central

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Metadata

Authors
Laffer, B., Bauer, D., Wasmuth, S., Busch, M., Jalilvand, T. V., Thanos, S., Meyer zu Hörste, G., Loser, K., Langmann, T., Heiligenhaus, A., & Kasper, M.
Affiliations
  • St. Franziskus Hospital (DE)
  • University of Münster (DE)
  • University of Cologne (DE)
Corresponding author
Kasper, Maren
Venue
Frontiers in Cellular Neuroscience
Published
2019-10-09
Volume
13
Pages
430-430
ISSN
1662-5102 (ISSN-L)
OpenAlex
W2979963107
Cited by
120
References
115
Citations by year
2026: 21 · 2025: 16 · 2024: 14 · 2023: 16 · 2022: 16 · 2021: 20 · 2020: 16
FWCI
5.72
Type
journal article
Language
en
Open access
Locations
  • Frontiers in Cellular Neuroscience (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Kölner Universitäts PublikationsServer (Universität zu Köln) (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
Deutsche Forschungsgemeinschaft
Topics
  • Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
  • Life Sciences › Immunology and Microbiology › Immunology › Immune cells in cancer
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Inflammasome and immune disorders
Keywords
IL-10, microglia, M1 macrophage phenotype, M2 phenotype, microglial polarization, lipopolysaccharide, IL-10 deficiency, CD86, CD206, inflammatory cytokines, TNF-α, interleukin-6, TGF-β1, chemokines, neuroinflammation

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