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Updated: Aug 11, 2026

Prediction of Red Blood Cell Antibody Significance Using the Monocyte-Macrophage Assay
Published on: February 7, 2025
AM-3K, an anti-macrophage antibody, recognizes CD163, a molecule associated with an anti-inflammatory macrophage
Yoshihiro Komohara1, Junko Hirahara, Tomohiro Horikawa
1Department of Cell Pathology, Graduate School of Medical Sciences, Kumamoto University 1-1-1, Honjo Kumamoto, Kumamoto 860-8556, Japan.
Abstract:
CD163 is a member of the scavenger receptor cysteine-rich superfamily restricted to the monocyte/macrophage lineage and is thought to be a useful marker for anti-inflammatory or alternatively activated macrophages. In this study we used mass spectrometric analysis to determine that the antigen recognized by the antibody AM-3K, which we previously generated as a tissue macrophage-specific monoclonal antibody, was CD163. An anti-inflammatory subtype of macrophages stimulated by dexamethasone or interleukin-10 showed strong reactivity for AM-3K and increased expression of CD163 mRNA. Immunohistochemical staining of routinely processed pathological specimens revealed that AM-3K recognized a specialized subpopulation of macrophages. In granulomatous diseases such as tuberculosis, sarcoidosis, or foreign body reactions, tissue macrophages around granulomas, but not component cells of the granulomas such as epithelioid cells and multinucleated giant cells, showed positive staining for AM-3K. In atherosclerotic lesions, scattered macrophages in diffuse intimal lesions were strongly positive for AM-3K, whereas foamy macrophages in atheromatous plaques demonstrated only weak staining. We therefore suggest that, in routine pathological specimens, AM-3K is a useful marker for anti-inflammatory macrophages because these cells can be distinguished from inflammatory or classically activated macrophages. Because AM-3K cross-reacts with macrophage subpopulations in different animal species including rats, guinea pigs, rabbits, cats, dogs, goats, pigs, bovine species, horses, monkeys, and cetaceans, it will have wide application for detection of CD163 in various animals.
Insights
The antibody AM-3K specifically identifies CD163, a marker for anti-inflammatory macrophages. This finding aids in distinguishing macrophage subtypes in various tissues and species for research and diagnostics.
Area of Science:
- Immunology
- Cell Biology
- Pathology
Background:
- CD163 is a scavenger receptor on monocytes/macrophages.
- It is considered a marker for anti-inflammatory or alternatively activated macrophages.
- A previously generated monoclonal antibody, AM-3K, was investigated for its target antigen.
Purpose of the Study:
- To identify the antigen recognized by the AM-3K antibody.
- To evaluate AM-3K as a marker for anti-inflammatory macrophages in pathological specimens.
- To assess the cross-reactivity of AM-3K in different animal species.
Main Methods:
- Mass spectrometric analysis to identify the AM-3K antigen.
- Stimulation of macrophages with dexamethasone or interleukin-10.
- Immunohistochemical staining of pathological specimens and animal tissues.
Main Results:
- Mass spectrometry confirmed AM-3K recognizes CD163.
- AM-3K showed strong reactivity with anti-inflammatory macrophages stimulated by IL-10 or dexamethasone.
- AM-3K identified specific macrophage subpopulations in granulomatous diseases and atherosclerotic lesions, and cross-reacted with CD163 in multiple animal species.
Conclusions:
- AM-3K is a reliable marker for CD163 and anti-inflammatory macrophages in routine pathological specimens.
- AM-3K can distinguish anti-inflammatory macrophages from classical inflammatory macrophages.
- The broad species cross-reactivity of AM-3K offers wide applications in veterinary and comparative immunology research.
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