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Fresh mouse peritoneal macrophages have low scavenger receptor activity
J G Kim1, C Keshava, A A Murphy
1Department of Gynecology and Obstetrics, School of Medicine, Emory University, Atlanta, GA 30322, USA.
Abstract:
Peritoneal macrophages are easily isolated by lavage, suggesting that they are either nonadherent or weakly adherent in situ. Cultured macrophages express class A scavenger receptors (SCR), which mediate Ca2+-independent adhesion in vitro. We examined fresh peritoneal macrophages from mice and from women with endometriosis to determine whether the adherence of these cells was associated with increased expression of class A SCR. Fresh human macrophages were not immunoreactive to SCR antibodies; however, SCR immunoreactivity increased with time in culture. Fresh mouse and human macrophages took up minimal amounts of 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine (DiI)-acetyl-low density lipoproteins (Ac-LDL), a class A SCR ligand. Murine macrophages in culture for 24-72 h internalized four times more Ac-LDL than fresh cells. Cells cultured for 2 days incorporated 3.2 times more [14C] oleate than freshly isolated cells (55.7 +/- 7.9 versus 17.6 +/- 3.0 nmol/mg cell protein). In contrast to SCR activity, mouse macrophage SCR mRNA expression was similar in freshly isolated macrophages and those cultured for 3 days. These results suggest that peritoneal macrophages express only low levels of SCR activity in situ and that posttranscriptional regulation after isolation leads to an increase in SCR activity that correlates with adherence of the macrophages in vitro.
Insights
Peritoneal macrophages show low scavenger receptor class A (SCR) activity in their natural state. Isolation and culture increase SCR activity, correlating with macrophage adherence in vitro.
Area of Science:
- Immunology
- Cell Biology
Background:
- Peritoneal macrophages are typically nonadherent or weakly adherent in situ.
- Cultured macrophages exhibit scavenger receptor class A (SCR) expression, mediating adhesion.
Purpose of the Study:
- To investigate the relationship between peritoneal macrophage adherence and SCR expression in fresh and cultured cells.
- To compare SCR expression in macrophages from mice and humans with endometriosis.
Main Methods:
- Immunoreactivity assays using SCR antibodies.
- Uptake studies with DiI-acetyl-low density lipoproteins (Ac-LDL), an SCR ligand.
- Measurement of [14C] oleate incorporation.
- Analysis of SCR mRNA expression.
Main Results:
- Fresh human macrophages showed minimal SCR immunoreactivity, which increased with culture time.
- Macrophages from both mice and humans exhibited low Ac-LDL uptake in situ, significantly increasing after 24-72 hours in culture.
- Cells cultured for 2 days showed increased oleate incorporation compared to fresh cells.
- SCR mRNA levels remained consistent between fresh and cultured mouse macrophages.
Conclusions:
- Peritoneal macrophages express low levels of SCR activity in situ.
- Posttranscriptional regulation following isolation leads to increased SCR activity and enhanced adherence in vitro.