Elsevier

Kidney International

Volume 67, Issue 6, June 2005, Pages 2488-2493
Kidney International

Technical Note
Antibodies against macrophages that overlap in specificity with fibroblasts

https://doi.org/10.1111/j.1523-1755.2005.00358.xGet rights and content
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Antibodies against macrophages that overlap in specificity with fibroblasts.

Background

Fibroblasts can be misidentified as macrophages because both cell types share antigens that are associated with popular antibodies targeting the monocyte/macrophage lineage. With the recent description of fibroblast-specific protein 1 (FSP1), we revisited the specificity of antibodies directed against macrophages to determine systematically which antibodies best distinguish both cell types in fibrotic tissues.

Methods

Tissue fibrosis was produced in mice carrying the GFP transgene encoding green fluorescent protein under the control of the FSP1 promoter. Single cell suspensions from these marked tissues were submitted for flow cytometry using antibodies against Mac-1, Mac-2, Mac-3, F4/80, CD68, major histocompatibility complex (MHC) class II, and CD45, and cDNA amplification of mRNA encoding the above target antigens was performed using specific primer sets in sorted pools of cells. Fibrotic tissues were also stained by immunohistochemistry with the same antibodies and examined under confocal microscopy.

Results

Comparison overlap between FSP1+ fibroblasts with each of the macrophage markers demonstrated that all antimacrophage antibodies (Mac-1, Mac-2, Mac-3, CD68, MHC class II, and CD45) except one (F4/80) recognize both cell types.

Conclusion

Antibodies directed against F4/80 clearly distinguish macrophages from FSP1+ fibroblasts in fibrotic tissues and is the preferred antibody in mice.

KEYWORDS

macrophage
fibroblast
FSP1
S100A4
F4/80
Mac-1
Mac-2
Mac-3
CD68
CD45

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