RnDSy-lu-2945 The Complex Biology of Macrophages: Origins, Functions, & Activation States

Common Macrophage Markers Models of Macrophage Activation B7-1/CD80 EMR1 (Human) LAMP2/CD107b Following development, macrophages encounter diverse stimuli, which can alter their B7-2/CD86 F4/80 (Mouse) LILRB4/CD85k/ILT3 The Origins of Mouse Tissue-Resident CCR5 Fcg RI/CD64 M-CSF R/CD115 transcriptional programs leading to an activated state. The classic view of macrophage CD11b/Integrin aM Fcg RII/CD32 MHC Class II Bone marrow activation is described in a dichotomous model. According to this model, classical (M1) CD11c Fcg RIII/CD16 Siglec-3/CD33 Yolk sac Macrophages Redefined Yolk sac-derived “Early” or “Late” g CD14 Galectin-3/Mac-2 TLR2 macrophage activation is induced by IFN- or lipopolysaccharide (LPS) and promotes a pro- E7.5 and Placenta Erythro-myeloid Progenitor Cell CD15 (SSEA-1)/Lewis X GITR Ligand TLR4 inflammatory response, while alternative (M2) macrophage activation is induced by IL-4, CD68/SR-D1 HLA-DR During development and throughout life, macrophages reside in many tissues of the body, contributing to both the E8.25 CD163 Integrin aL/CD11a IL-10, or IL-13 and stimulates an anti-inflammatory response. Slight phenotypic variations maintenance of tissue homeostasis and the immune response following injury or pathogenic insult. In the late 1960s, van noted in the M2 phenotype, subsequently led to the description of different M2 subsets Furth and Cohn proposed that tissue-resident macrophages develop primarily from circulating, bone marrow-derived blood known as M2a, M2b, M2c, and M2d, which were defined based on the stimuli used for monocytes. This model was widely accepted until recent fate-mapping studies demonstrated that several tissue-resident Common Macrophage Functions activation (top below). It has been widely recognized however, that even this expanded model macrophage populations in mice arise from HSC-independent embryonic precursors and are maintained by self-renewal. of macrophage activation is too simplistic to account for the range of phenotypes observed Yolk sac-derived The earliest macrophages or primitive progenitors arise from early and late erythro-myeloid progenitors (EMPs) generated in macrophages under different homeostatic and pathological conditions. For this reason, a Immune Surveillance Primitive Macrophage in the extra-embryonic yolk sac during primitive hematopoiesis at embryonic age 7.5 and 8.25 (E7.5 and E8.25). These Around E8.5 new multidimensional model of macrophage activation has recently been proposed based on EMPs can give rise to yolk sac-derived macrophages without passing through a monocytic intermediate and are the first to extensive gene expression analysis (bottom below). This model suggests that a spectrum of TLR seed the fetal tissues following initiation of the blood circulation. With the exception of microglial cells in the brain, the activation states spanning the M1/M2 states can occur in response to diverse signals primitive macrophages in most fetal tissues are subsequently replaced either partially or completely by fetal liver-derived including ontogeny-related signals, tissue-specific signals, and stress signals, which are E17.5-Adult monocytes. Fetal liver monocytes are generated from EMPs derived from either the yolk sac or hemogenic endothelium of integrated to determine the macrophage response. Yolk sac-derived “Late” the placenta and umbilical cord, or from hematopoietic stem cells (HSCs) generated in the para-aortic splanchnopleura Erythro-myeloid (P-Sp) and aorta-gonad-mesonephros (AGM) regions of the embryo. These progenitors migrate to the fetal liver in two Progenitor Cell Fetal liver HSC Granulocyte-Macrophage Macrophage-Dendritic Classical/Alternative Model of Macrophage Activation TLR TLR E9.5 successive waves around E9.5 (EMPs) and E10.5/E11 (immature and mature HSCs) and expand, giving rise to fetal liver Myeloid from P-Sp or AGM Progenitor Progenitor Pro-Inflammatory Pro-Inflammatory monocytes, which enter the circulation and differentiate into macrophages in peripheral tissues. In some tissues, including M1 IFN-γ, LPS, OxLDL, Progenitor Cholesterol crystals CXCL M4 Macrophages the liver, lung, skin, spleen, and peritoneum, fetal liver monocyte-derived macrophages maintain the ability to self-renew Cell Common Monocyte Hemoglobin/ Haptoglobin Complex Progenitor (cMoP) Heme into adulthood and establish the tissue-resident population. In other tissues, such as the dermis and gut, fetal liver HSC monocyte-derived macrophages are gradually replaced by the recruitment of bone marrow-derived monocytes generated P-Sp or AGM cMoP IL-, IL-13 Common Dendritic HSC Progenitor HSC OxPL Phagocytosis M2a Phagosome from adult hematopoiesis beginning around E17.5. Immune Complex Mhem s Progenitor (CDP) e + IL-1 R/TLR Ligands p y Removal of: Pathogen E10.5 t E10.5 b IL-10, TGF-β, u • Dead/Dying cells/Debris Glucocorticoid Hormones S TLR Agonists + Tissue-resident macrophages are a versatile, heterogeneous group of cells that support multiple tissue functions. Most of Adenosine M(Hb) ic • Pathogens if c e • Toxins Lysosome Phagolysosome Fetal Liver- p our knowledge about these cells has come from studies in mice, which suggest that the phenotypes and functional programs -s M2b is derived Monocyte os er of tissue macrophages are determined by signals that they receive in their tissue microenvironments. Aside from providing E11.5–E12.5 scl Blood Monocytes Mox ero M2d Ath Pathogen Destruction the first line of defense against invading pathogens, tissue-resident macrophages have a fundamental role in maintaining M2c Wound Healing & Anti-Inflammatory Anti-Inflammatory Angiogenesis Anti-oxidant Atheroprotective tissue integrity and homeostasis. In addition, they may have specialized functions based on their locations and distinct E13.5–E14.5 E13.5–E14.5 gene expression profiles. For example, osteoclasts are bone-resident macrophages that specialize in bone resorption, while M1 Markers red pulp macrophages in the spleen specialize in heme degradation and iron recycling. Abnormalities in macrophage E14.5–E18.5 Fetal Liver-derived Monocyte Cell Surface Markers Intracellular Markers Secreted Proteins Antigen Presentation Antigen functions have been associated with a wide range of chronic inflammatory and autoimmune diseases including obesity and B7-1/CD80+ COX2+ Cytokines Chemokines Lysosome T Cell B7-2/CD86+ iNOS+ IL-1b+ CCL2−CCL5+ Antigen type II diabetes, asthma, atherosclerosis, fibrosis, cancer, inflammatory bowel disease, multiple sclerosis, and rheumatoid CD36/SR-B3+ IRF5+ IL-6+ CCL8+ Class II MHC arthritis, suggesting that macrophages may serve as therapeutic targets. This possibility, however, requires a greater CD68/SR-D1+ STAT1+ IL-12high CCL11+ Tissue-resident Recruited Inflammatory Macrophage CD163− IL-15+ CCL15+ (Human) + + + MHC TCR-CD3 understanding of the differences in the development, phenotypes, and functions of tissue-resident macrophages. Macrophage Fcg RII/CD32 M1 Macrophage Functions: IL-17 CCL19 Fcg RIII/CD16+ Defense against pathogens IL-18+ CCL20+ + CD28 IFN-g R+ IL-23+ CXCL1−3 M1-Associated Pathologies: + Tissue MHC class IIhigh IFN-g+ CXCL5 B7-1/CD80 Chronic inflammation, tissue damage, + TNF-a+ CXCL8−11 autoimmune disease CXCL13+ CXCL16+ Phagolysosome Markers, Origins, & Specialized Functions of Select Mouse Tissue-resident Macrophages CX3CL1+ Cytokine Production Ligand M2a Markers M2b Markers M2c Markers TLR Adipose Tissue Blood Bone Central Nervous System Gastrointestinal Tract Cell Surface Markers Cell Surface Markers Cell Surface Markers Ligand CD163+ B7-2/CD86+ CCR2+ + + + Perivascular Macrophage Meningeal Macrophage Microglia Dietary Antigens Commensal Bacteria Intestinal lamina propria macrophages CD200 R1 IL-4 Ra CD150/SLAM low Markers: Markers: + + + NLR Ly-6C Monocytes CLEC10A/CD301 MHC class II CD163 + a + + a+ Osteoclasts AIF-1/Iba1 CD11b/Integrin M CXCR1 Intracellular Markers IL-4 R Osteoclast a + low/int + + Bone Marrow CD11b/Integrin M CD11c CXCR2 COX2+ MMR/CD206 Markers: low high + + Macrophage + + CD45 CX3CR1 DC-SIGN/CD209 IRF4 SR-AI/MSR Calcitonin Receptor + + + + low/− F4/80 F4/80 Dectin-1 SOCS3+ SR-B1 F4/80 + + + + Fcg RI/CD64 Fce RIa + TLR1 low RANK Perivascular macrophages Sphingosine Kinase 1/2 Adipose tissue-associated Ly-6C monocytes MHC Class IIhigh IL-1 RII+ NFATc1+ Intracellular Markers Cytokines Markers: low Secreted Proteins macrophages Markers: MHC class II + Tartrate-resistant acid phosphatase (TRAP)+ + Arginase 1 (Mouse) Markers: high AIF-1/Iba1 + Cytokines/Growth Factors CD11b/Integrin aM MMR/CD206 + Perivascular Space + + IRF4 CD45+ + CD11b/Integrin aM G-CSF CD43 Bone marrow macrophages + high Intracellular Markers + SOCS3 + + CD45 Intestinal lamina propria macrophages GM-CSF F4/80 CX3CR1 Markers: + + + + Arginase 1 (Mouse) + TLR8 g + + CD163 IL-1b PPAR F4/80 CD169 Origin: Bone marrow-derived monocytes + + IRF4 + low + Microglia F4/80 IL-6 Secreted Proteins Ly-6C ER-HR3 Specialized Functions: + + Maintenance of PPARg + + + MMR/CD206 IL-10 Cytokines/Growth Factors Wound Healing and Tissue Remodeling M-CSF R/CD115 F4/80 intestinal homeostasis; Recognition and + + STAT6 + + + − SR-AI/MSR TNF-a IL-10 Inflammatory: NGFI-Ba/Nur77/NR4A1 Tartrate-resistant acid phosphatase (TRAP) removal of intestinal pathogens; + + Secreted Proteins Chemokines TGF-b • Phagocytosis Tissue Injury VCAM-1/CD106 Brain Parenchyma Meningeal macrophages Maintenance of gut epithelial integrity Intestinal Lamina Cytokines/Growth Factors CCL1+ Chemokines • Antigen Presentation Markers: Osteoblasts Propria Macrophages Associated Deficiencies or Pathologies: FIZZ1/RELMa+ (Mouse) CCL20+ CCL16+ • Pro-inflammatory Cytokine Production + AIF-1/Iba1 Inflammatory bowel disease high + + a + IL-1ra CXCL1−CXCL3 CCL18 (Human) CD11b/Integrin M high + Proliferative: Adipose Tissue Macrophage high IL-10 CXCL13 CD45 low • Fibroblast Recruitment/Activation + IL-12 low Osteoclasts Bone marrow macrophages F4/80 + • ECM Formation Matrix Production Adipose tissue-associated macrophages Ly6C monocytes TGF-b • Reepithelialization Origin: Presumed to be bone marrow- Origin: Yolk sac-derived or fetal liver- Chemokines MMPs Origin: Not clear Origin: Bone marrow-derived monocytes • Neovascularization/Angiogenesis derived monocytes derived monocytes Microglia Perivascular macrophages Spleen CCL1+ M2 Macrophage Functions: Fibroblasts Specialized Functions: Adipogenesis; Adaptive thermogenesis; Regulation of insulin Specialized Functions: Immune surveillance; Maintenance of vascular integrity Red pulp macrophages + Specialized Functions: Specialized Functions: CCL2 Wound healing & Tissue remodeling Remodeling: Bone resorption and Support erythropoeisis; Origin: Yolk sac-derived Origin: Bone marrow-derived monocytes Markers: Growth Factors sensitivity and glucose tolerance Red Pulp Marginal Zone CCL14+ (Human) • ECM Composition Modifications remodeling; Maintenance of the HSC niche Maintenance of the HSC niche Macrophage a low M2-Associated Pathologies: Associated Deficiencies or Pathologies:Obesity, insulin resistance, diabetes, loss of Specialized Functions: Brain development; Provision Specialized Functions: Immune surveillance Macrophage CD11b/Integrin M CCL17+ • ECM Protein Synthesis Associated Deficiencies or Pathologies: Associated Deficiencies or Pathologies:Disruption + Cancer, fibrosis, epithelial hyperplasia Enzymes adaptive thermogenesis of neurotrophic factors; Removal of dead neurons CD36/SR-B3 CCL18+ Lung Osteopetrosis, osteoporosis, arthritis of hematopoiesis Meningeal macrophages + Cytokines Macrophages Alveolar macrophages and synaptic pruning CD68/SR-D1 CCL22+ Markers: Origin: Bone marrow-derived monocytes + Associated Deficiencies or Pathologies: CD163 CCL23+ (Human) CD11b/Integrin aMlow/– + Neurodegeneration Specialized Functions: Immune surveillance Dectin-2 CCL24+ CD11chigh + Liver Red F4/80 CCL26+ CD68/SR-D1+ Serosal Tissues + Pleural macrophages Pulp MMR/CD206 YM1/Chitinase 3-like 3+ (Mouse) Kupffer cells CD200 R1+ SIRPa/CD172a+ Tumor Infiltration Hepatocytes Sinusoidal Cell Pleural Macrophage Markers: Markers: Alveolar DEC–205/CD205int Marginal Spi-C+ CD11b/Integrin aMhigh Skin Langerhans cells Zone B7-1/CD80low/– Macrophages Dectin-1+ + F4/80high Markers: VCAM-1/CD106 M2d Markers low low CD11b/Integrin aM F4/80 + Tim-4+ CD11b/Integrin aM + + Marginal zone macrophages CD68/SR-D1 Galectin-3/Mac-2 Langerhans Cell CD11c+ Intracellular Markers Secreted Proteins high + Markers: + F4/80 MARCO Large peritoneal macrophages + Chemokines F4/80 + iNOS Cytokines/Growth Factors + low CD68/SR-D1 + + Galectin-3/Mac-2 MHC class II Markers: + CCL5 Id2 + IL-10 + high high White Dectin-2 low + Kupffer Cell PPARd MMR/CD206 CD11b/Integrin aM + IL-12 CXCL10 Langerin Pulp F4/80low Siglec-1/CD169+ g+ CD11clow low + M1 M2 PPAR + + TNF-a CXCL16 Tumor Rejection Tumor Escape – RUNX3/CBFA3 LXRa/NR1H3 Siglec-Fhigh DC-SIGN/CD209 + Pleural + VEGF Cavity F4/80high Dermal macrophages Marginal MARCO Hepatic Sinusoid Motile liver macrophages Epidermis + Large Peritoneal Small Peritoneal GATA-6+ Markers: White Pulp Metallophilic SIGNR1/CD209b Markers: + Interstitial macrophages Peritoneal Macrophage Macrophage low/– a + Macrophage Macrophage TIM-4 high MHC class II CD11b/Integrin M B7-1/CD80 Markers: Cavity – low + L-Selectin/CD62L CD11c Marginal metallophilic macrophages New Multidimensional Model of Macrophage Activation CD11b/Integrin aM CD11b/Integrin aMint Interstitial TIM-4+ CLEC10A/CD301+ + – Markers: F4/80 Macrophages CD11c + Red pulp macrophages Dectin-1 + + CD68/SR-D1 Inflammatory Monocytes CD68/SR-D1 Small peritoneal macrophages + Origin: Fetal liver-derived monocytes Ontogeny-related Dermis Dectin-2 F4/80low CD200 R1+/– Markers: Signals Tissue-specific Signals F4/80+ Specialized Functions: Clearance of aged or damaged + + low LXRa/NR1H3 F4/80 CD11b/Integrin aM high Fcg RI/CD64 erythrocytes; Capture of blood-borne debris; Siglec-1/CD169+ Motile Liver MHC class II+/– CD11c– MerTK+ Heme degradation; Iron recycling Tissue-resident Stellate Cell Macrophage Siglec-F– DC-SIGN/CD209+ Cytotoxicity MHC class IIlow White pulp (tingible body) macrophages Macrophage • Direct • Antibody-dependent Cell-mediated Cytotoxicity low Associated Deficiencies or Pathologies:Impaired iron Alveolar macrophages F4/80 Dermal Macrophage + Markers: high MMR/CD206 homeostasis and erythrocyte clearance MHC class II + Stress Signals: Cytokines, Growth Factors, Damage-associated Origin: Fetal liver-derived monocytes Siglec-1/CD169high CD68/SR-D1 TNF Target Cell Kupffer cells Motile liver macrophages Pleural macrophages L-Selectin/CD62L+ + Molecular Patterns, Pathogen-associated Molecular Patterns, TNF-α Specialized Functions: Recycling of surfactant molecules; Immune surveillance of inhaled pathogens; Marginal zone macrophages MFG-E8 Granzymes Origin: Yolk sac-derived or fetal liver- Origin: Bone marrow-derived monocytes Origin: TIM-4– + Eicosanoids, Leukotrienes, Fatty Acids, Cholesterol, Other Signals NO Clearance of allergens, dust, and microorganisms Bone marrow-derived monocytes Origin: Bone marrow-derived monocytes MerTK + Activation States derived monocytes Specialized Functions: Immune surveillance Specialized Functions: Langerhans cells Dermal macrophages TIM-4 Oxygen Associated Deficiencies or Pathologies:Pulmonary alveolar proteinosis Immune surveillance Specialized Functions: Clearance of pathogens present in the A spectrum of Specialized Functions: Clearance of aged Radicals Associated Deficiencies or Pathologies:Hepatic Origin: Yolk sac-derived or fetal liver- Origin: Bone marrow-derived monocytes circulation; Retention of marginal zone B cells activation states with erythrocytes, blood-borne particles, toxins, and Interstitial macrophages Large peritoneal macrophages fibrosis, fatty liver disease, non-alcoholic derived monocytes Specialized Functions: Immune surveillance different cell surface microorganisms; Tissue homeostasis and Origin: Yolk sac-derived or fetal liver-derived monocytes Marginal metallophilic macrophages Antigen steatohepatitis Origin: Fetal liver- and bone marrow-derived monocytes Specialized Functions: Immune surveillance and intracellular M1 M2 remodeling in the liver; Metabolic function Specialized Functions: Regulation of IgA production in the gut by peritoneal B1 cells Origin: Bone marrow-derived monocytes markers depending Specialized Functions: Regulation of dendritic cell function Associated Deficiencies or Pathologies:Defects Fc Receptors in the liver on the stimuli Small peritoneal macrophages in wound healing, fibrosis Specialized Functions: Clearance of pathogens present Target Associated Deficiencies or Pathologies:Hepatic in the circulation Origin: Bone marrow-derived monocytes Pro-Inflammatory Anti-Inflammatory Cell Lysis fibrosis, fatty liver disease, non-alcoholic White pulp macrophages Macrophage steatohepatitis, impaired erythrocyte clearance Specialized Functions: Immune surveillance Specialized Functions: Clearance of apoptotic B cells

NOTE: This poster conveys a general overview and should be considered neither comprehensive nor definitive. The details of this information are understood to be subject to interpretation. © R&D Systems 2016

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