AM-3K, an Anti-macrophage Antibody, Recognizes CD163, a Molecule Associated with an Anti-inflammatory Macrophage Phenotype

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.

[1]  Tudung T Nguyen,et al.  Expression of CD163 (Hemoglobin Scavenger Receptor) in Normal Tissues, Lymphomas, Carcinomas, and Sarcomas Is Largely Restricted to the Monocyte/Macrophage Lineage , 2005, The American journal of surgical pathology.

[2]  W. Brown,et al.  Studies on translocation of immunoglobulins across intestinal epithelium , 1977, The American Journal of Digestive Diseases.

[3]  S Gordon,et al.  Macrophage receptors and immune recognition. , 2005, Annual review of immunology.

[4]  Silvano Sozzani,et al.  The chemokine system in diverse forms of macrophage activation and polarization. , 2004, Trends in immunology.

[5]  P. Chu,et al.  CD163: a specific marker of macrophages in paraffin-embedded tissue samples. , 2004, American journal of clinical pathology.

[6]  L. Boon,et al.  Gaucher cells demonstrate a distinct macrophage phenotype and resemble alternatively activated macrophages. , 2004, American journal of clinical pathology.

[7]  P. Guyre,et al.  Cross‐linking of FcγR triggers shedding of the hemoglobin‐haptoglobin scavenger receptor CD163 , 2004, Journal of leukocyte biology.

[8]  P. Picci,et al.  Proteases and interleukin-6 gene analysis in 92 giant cell tumors of bone. , 2004, Annals of oncology : official journal of the European Society for Medical Oncology.

[9]  P. Högger,et al.  Biochemical Characterization of a Glucocorticoid-Induced Membrane Protein (RM3/1) in Human Monocytes and Its Application as Model System for Ranking Glucocorticoid Potency , 1998, Pharmaceutical Research.

[10]  M. Takeya,et al.  A novel monoclonal antibody, RM-4, specifically recognizes rat macrophages and dendritic cells in formalin-fixed, paraffin- embedded tissues , 1997, The Histochemical Journal.

[11]  M. Takeya,et al.  Immunohistochemical detection of macrophages in the short-finned pilot whale (Globicephala macrorhynchus) and Risso's dolphin (Grampus griseus). , 2004, Journal of comparative pathology.

[12]  Y. Yoo,et al.  Immunohistochemical Characterization of Macrophage and Dendritic Cell Subpopulations of the Spleen, Thymus, Tongue and Heart in Cyclophosphamide‐induced Immunosuppressed Rat , 2003, Anatomia, histologia, embryologia.

[13]  D. Mosser,et al.  The many faces of macrophage activation , 2003, Journal of leukocyte biology.

[14]  Joel V Weinstock,et al.  Granulomas in schistosome and mycobacterial infections: a model of local immune responses. , 2003, Trends in immunology.

[15]  S. Gordon Alternative activation of macrophages , 2003, Nature Reviews Immunology.

[16]  P. Wallace,et al.  Endotoxin induces rapid metalloproteinase‐mediated shedding followed by up‐regulation of the monocyte hemoglobin scavenger receptor CD163 , 2002, Journal of leukocyte biology.

[17]  B. Ohlsson,et al.  Oxysterols induce interleukin‐1β production in human macrophages , 2002, European journal of clinical investigation.

[18]  G. Amphlett,et al.  Distribution of IFN-gamma, IL-4 and TNF-alpha protein and CD8 T cells producing IL-12p40 mRNA in human lung tuberculous granulomas. , 2002, Immunology.

[19]  D. Schaer,et al.  Molecular cloning and characterization of the mouse CD163 homologue, a highly glucocorticoid-inducible member of the scavenger receptor cysteine-rich family , 2001, Immunogenetics.

[20]  S. Moestrup,et al.  Identification of the haemoglobin scavenger receptor , 2001, Nature.

[21]  M. Takeya,et al.  Immunohistochemical analysis of macrophages and myofibroblasts appearing in hepatic and renal fibrosis of dogs. , 2001, Journal of comparative pathology.

[22]  R. Hernández-Pando,et al.  Expression of inducible nitric oxide synthase and nitrotyrosine during the evolution of experimental pulmonary tuberculosis. , 2001, Experimental and toxicologic pathology : official journal of the Gesellschaft fur Toxikologische Pathologie.

[23]  P. Wallace,et al.  Human monocytes express CD163, which is upregulated by IL-10 and identical to p155. , 2000, Cytokine.

[24]  M. Takeya,et al.  Distribution of Cells Immunopositive for AM-3K, a Novel Monoclonal Antibody Recognizing Human Macrophages, in Normal and Diseased Tissues of Dogs, Cats, Horses, Cattle, Pigs, and Rabbits , 2000, Veterinary pathology.

[25]  T. Langmann,et al.  Regulation of scavenger receptor CD163 expression in human monocytes and macrophages by pro‐ and antiinflammatory stimuli , 2000, Journal of leukocyte biology.

[26]  R. Beelen,et al.  Regulation of CD163 on human macrophages: cross‐linking of CD163 induces signaling and activation , 1999, Journal of leukocyte biology.

[27]  U. Lalloo,et al.  Nitric oxide levels in exhaled air and inducible nitric oxide synthase immunolocalization in pulmonary sarcoidosis. , 1999, The European respiratory journal.

[28]  S. Goerdt,et al.  Other functions, other genes: alternative activation of antigen-presenting cells. , 1999, Immunity.

[29]  W. Sanger,et al.  Transcriptional regulation of MMP-9 expression in stromal cells of human giant cell tumor of bone by tumor necrosis factor-alpha. , 1999, International Journal of Oncology.

[30]  W. Sanger,et al.  Interleukin-1β Upregulates MMP-9 Expression in Stromal Cells of Human Giant Cell Tumor of Bone , 1999 .

[31]  P. Högger,et al.  Identification of the integral membrane protein RM3/1 on human monocytes as a glucocorticoid-inducible member of the scavenger receptor cysteine-rich family (CD163). , 1998, Journal of immunology.

[32]  Z. Yasruel,et al.  Cytokine mRNA gene expression in active and nonactive pulmonary sarcoidosis. , 1997, The European respiratory journal.

[33]  J. Möst,et al.  Formation of multinucleated giant cells in vitro is dependent on the stage of monocyte to macrophage maturation. , 1997, Blood.

[34]  M. Takeya,et al.  Interspecies reactivities of anti-human macrophage monoclonal antibodies to various animal species. , 1996, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[35]  M. Takeya,et al.  AM‐3K, A NOVEL MONOCLONAL ANTIBODY SPECIFIC FOR TISSUE MACROPHAGES AND ITS APPLICATION TO PATHOLOGICAL INVESTIGATION , 1996, The Journal of pathology.

[36]  I. Cree,et al.  Detection of tumour necrosis factor alpha in sarcoidosis and tuberculosis granulomas using in situ hybridisation. , 1994, Journal of clinical pathology.

[37]  D. Mason,et al.  A new macrophage differentiation antigen which is a member of the scavenger receptor superfamily , 1993, European journal of immunology.

[38]  P. Tipping,et al.  Production of tumor necrosis factor and interleukin-1 by macrophages from human atheromatous plaques. , 1993, The American journal of pathology.

[39]  S. McCarthy,et al.  A monocyte/macrophage antigen recognized by the four antibodies GHI/61, Ber-MAC3, Ki-M8 and SM4. , 1992, Immunology.

[40]  H. Stein,et al.  Ber-MAC3: new monoclonal antibody that defines human monocyte/macrophage differentiation antigen. , 1991, Journal of clinical pathology.

[41]  A. Mackensen,et al.  Microculture assay for human macrophage maturation in vitro. Cell-ELISA analysis of differentiation antigen expression. , 1988, International Archives of Allergy and Applied Immunology.

[42]  H. Radzun,et al.  Ki-M8 monoclonal antibody reactive with an intracytoplasmic antigen of monocyte/macrophage lineage. , 1987, Blood.

[43]  P. Nakane,et al.  Studies on translocation of immunoglobulins across intestinal epithelium. II. Immunoelectron-microscopic localization of immunoglobulins and secretory component in human intestinal mucosa. , 1976, Gastroenterology.