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Published on: May 19, 2023
Regulation of CD163 mRNA and soluble CD163 protein in human adipose tissue in vitro
Karen Fjeldborg1, Holger J Møller2, Bjørn Richelsen2
1Departments of Endocrinology and Internal MedicineMEAClinical BiochemistryAarhus University Hospital, Tage Hansensgade 2, DK-8000 Aarhus C, Denmark Karen.fjeldborg@ki.au.dk.
Abstract:
CD163-positive macrophages are highly expressed in the human adipose tissue (AT) particularly from obese individuals. However, little is known about the regulation of CD163 mRNA and the protein level of sCD163 in human AT. We aimed to examine the regulation of CD163 and sCD163 in AT. Human s.c. AT samples (n=5) were stimulated with dexamethasone (DEX; 200 nmol/l), lipopolysaccharide (LPS; 100 ng/ml), or DEX+LPS for various time periods up to 24 h. Gene expressions of CD163, ADAM17, IL10, and TNFA (TNF) were measured by RT-PCR. Protein levels of sCD163, IL10, and TNFα (TNF) were measured by ELISA. Furthermore, AT was separated into stromal and adipocyte fraction. We found that CD163 mRNA was strongly expressed in the stromal vascular fraction but hardly detectable in the isolated adipocytes. Incubating whole AT with DEX significantly up-regulated CD163 (P<0.001), whereas incubation with LPS had no effects on CD163 (P>0.05). By contrast, the protein level of sCD163 was not affected by DEX (P>0.05), but LPS significantly increased the level of sCD163 and TNFα (P<0.05). This might be due to the concomitant LPS stimulation of ADAM17, which is known to mediate shedding of the extracellular domains of sCD163 and TNFα. Finally, DEX significantly reduced the LPS-induced TNFα release to the incubation medium but had no effects on sCD163. We conclude that the expression of CD163 and the release of sCD163 are differentially regulated in human AT. Moreover, similar to studies on differentiated blood monocytes, TNFα and sCD163 are concomitantly released in human AT by LPS, which also up-regulate ADAM17.
Insights
Regulation of CD163 and soluble CD163 (sCD163) in human adipose tissue differs. Dexamethasone increases CD163 mRNA, while lipopolysaccharide boosts sCD163 and TNFα protein levels.
Area of Science:
- Immunology
- Adipose Tissue Biology
- Macrophage Biology
Background:
- CD163-positive macrophages are prevalent in human adipose tissue, especially in obesity.
- Understanding the regulation of CD163 mRNA and soluble CD163 (sCD163) protein in adipose tissue is limited.
Purpose of the Study:
- To investigate the differential regulation of CD163 and sCD163 in human subcutaneous adipose tissue.
- To explore the effects of dexamethasone (DEX) and lipopolysaccharide (LPS) on CD163 expression and sCD163 shedding.
Main Methods:
- Human subcutaneous adipose tissue samples were stimulated with DEX, LPS, or both.
- Gene expression of CD163, ADAM17, IL10, and TNFα was analyzed using RT-PCR.
- Protein levels of sCD163, IL10, and TNFα were quantified via ELISA.
- Adipose tissue was fractionated into stromal and adipocyte components.
Main Results:
- CD163 mRNA was primarily found in the stromal vascular fraction, not isolated adipocytes.
- DEX significantly upregulated CD163 mRNA, while LPS had no effect.
- LPS, but not DEX, significantly increased sCD163 and TNFα protein levels.
- LPS induced ADAM17 expression, potentially mediating the shedding of sCD163 and TNFα.
- DEX reduced LPS-induced TNFα release but did not affect sCD163 levels.
Conclusions:
- CD163 expression and sCD163 release are distinctly regulated in human adipose tissue.
- LPS stimulation leads to the release of TNFα and sCD163, alongside ADAM17 upregulation, mirroring findings in monocytes.
- These findings highlight the complex interplay of immune regulation within adipose tissue, particularly in response to inflammatory stimuli.
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