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Updated: Feb 20, 2026
Isolation, Characterization, and Purification of Macrophages from Tissues Affected by Obesity-related Inflammation
Published on: April 3, 2017
Activated macrophages control human adipocyte mitochondrial bioenergetics via secreted factors
Michaela Keuper1, Stephan Sachs2, Ellen Walheim3
1Institute of Diabetes and Regeneration Research, Helmholtz Zentrum Muenchen, German Research Center for Environmental Health (GmbH), Neuherberg, Germany; German Center for Diabetes Research (DZD), Neuherberg, Germany; Institute of Experimental Genetics, Helmholtz Zentrum Muenchen, German Research Center for Environmental Health (GmbH), Neuherberg, Germany.
Objective:
Obesity-associated WAT inflammation is characterized by the accumulation and local activation of macrophages (MΦs), and recent data from mouse studies suggest that macrophages are modifiers of adipocyte energy metabolism and mitochondrial function. As mitochondrial dysfunction has been associated with obesity and the metabolic syndrome in humans, herein we aimed to delineate how human macrophages may affect energy metabolism of white adipocytes.
Methods:
Human adipose tissue gene expression analysis for markers of macrophage activation and tissue inflammation (CD11c, CD40, CD163, CD206, CD80, MCP1, TNFα) in relationship to mitochondrial complex I (NDUFB8) and complex III (UQCRC2) was performed on subcutaneous WAT of 24 women (BMI 20-61 kg/m2). Guided by these results, the impact of secreted factors of LPS/IFNγ- and IL10/TGFβ-activated human macrophages (THP1, primary blood-derived) on mitochondrial function in human subcutaneous white adipocytes (SGBS, primary) was determined by extracellular flux analysis (Seahorse technology) and gene/protein expression.
Results:
Stepwise regression analysis of human WAT gene expression data revealed that a linear combination of CD40 and CD163 was the strongest predictor for mitochondrial complex I (NDUFB8) and complex III (UQCRC2) levels, independent of BMI. IL10/TGFβ-activated MΦs displayed high CD163 and low CD40 expression and secreted factors that decreased UQCRC2 gene/protein expression and ATP-linked respiration in human white adipocytes. In contrast, LPS/IFNγ-activated MΦs showed high CD40 and low CD163 expression and secreted factors that enhanced adipocyte mitochondrial activity resulting in a total difference of 37% in ATP-linked respiration of white adipocytes (p = 0.0024) when comparing the effect of LPS/IFNγ- vs IL10/TGFβ-activated MΦs.
Conclusion:
Our data demonstrate that macrophages modulate human adipocyte energy metabolism via an activation-dependent paracrine mechanism.
Insights
Human macrophages influence white adipocyte energy metabolism. Macrophage activation state dictates whether they enhance or decrease adipocyte mitochondrial function, impacting obesity and metabolic syndrome.
Area of Science:
- Cell Biology
- Metabolic Syndrome Research
- Immunology
Background:
- Obesity-associated adipose tissue inflammation involves macrophage accumulation and activation.
- Mitochondrial dysfunction is linked to human obesity and metabolic syndrome.
- Macrophages may influence adipocyte energy metabolism and mitochondrial function.
Purpose of the Study:
- To investigate how human macrophages affect the energy metabolism of white adipocytes.
- To determine the role of macrophage activation state in modulating adipocyte mitochondrial function.
Main Methods:
- Gene expression analysis of human adipose tissue for macrophage activation and inflammation markers.
- Assessing mitochondrial complex I and III levels in relation to macrophage markers.
- Evaluating the impact of activated human macrophages on white adipocyte mitochondrial function using Seahorse technology and gene/protein expression analysis.
Main Results:
- Macrophage markers CD40 and CD163 predict mitochondrial complex levels in adipose tissue, independent of BMI.
- IL10/TGFβ-activated macrophages (high CD163, low CD40) reduced adipocyte mitochondrial respiration.
- LPS/IFNγ-activated macrophages (high CD40, low CD163) enhanced adipocyte mitochondrial activity, showing a 37% difference in ATP-linked respiration compared to IL10/TGFβ-activated macrophages.
Conclusions:
- Human macrophages modulate white adipocyte energy metabolism through activation-dependent paracrine signaling.
- The specific activation state of macrophages determines their effect on adipocyte mitochondrial function.
- These findings highlight a novel mechanism linking immune cells to metabolic regulation in obesity.
