M1 macrophages express specific phenotype markers, including. Concomitantly, it is also necessary to understand how the in vitro culture conditions used to generate macrophages affect the expression of various surface markers. Elevated proinflammatory cytokine levels and markers of m1 macrophages are found in ppar. Monocytes and tissue macrophages in their various forms see table 1221 on the nelson textbook of pediatrics website at. Macrophages, by definition, are a type of immune cells capable of engulfing and. Understanding the mysterious m2 macrophage through. Related surface markers for m2type cells also contain cd68. Macrophage activation markers view the complex biology of macrophages poster showing the expression levels of different macrophage markers following development, macrophages encounter diverse stimuli, which can alter their transcriptional programs leading to an activated state. Moreover, fatty acid oxidation fao not only supports antiinflammatory responses as described formerly but also.
There are very few unique macrophage markers and often a number of markers will be required to identify your cell type. When a bcell in the lymph node recognizes the same unprocessed surface antigen on the bacterium with its surface bound antibody, the antigen is endocytosed and processed. Compared with marker cd68, cd163 is more selective to macrophages, so cd163 can be used as a highly specific marker for m2type macrophages 19. To provide a foundation for the complexity of in vivo macrophage phenotypes, we. Mitochondrial dysfunction prevents repolarization of. Flow cytometry and highcontent imaging to identify markers. Increase in circulating cells coexpressing m1 and m2. These markers are transmembrane glycoproteins, scavenger receptors, enzymes, growth. Intriguingly, we observed significant fpr2alx surface expression in both unstimulated human pbmcderived macrophages and m1phenotype cells, yet cell surface expression of the receptor was lost on cells stimulated toward an m2 phenotype with il4 supplemental figure 6. Here we chose cd40, a macrophage surface receptor involved in proinflammatory pathways, whose cell surface expression is increased in blood monocytederived macrophages induced by interferon. Previous human macrophage studies particularly focused on the differential gene expression profiles of m1 and m2 macrophages as well as on identification of functional surface markers 10, 20, 2729. Cd206, the mannose receptor 1, is a ctype lectin mainly present in macrophages and immature dc cells. A systematic approach to identify markers of distinctly activated.
According to this model, classical m1 macrophage activation is induced by ifngamma or lipopolysaccharide lps and promotes a proinflammatory response, while alternative m2 macrophage activation is induced by il4, il10, or il and stimulates an antiinflammatory response. Figure 3 the expression of macrophage markers in small macrophages compared to. Plasticity in the macrophage activation states mapping. Monocytes and macrophages are phagocytes, acting in both nonspecific defence or innate immunity as well as specific defense or cell mediated immunity of vertebrate. The alternatively activated or m2 macrophages are immune cells with high phenotypic heterogeneity and are governing functions at the interface of immunity, tissue homeostasis, metabolism, and endocrine signaling. These immune cells play key roles in controlling tissue homeostasis, infections, and excessive inflammation.
From these, expression of 9 surface markers were consistently altered by all conditioning regimens, while 9 were specific to individual polarizing stimuli. More recently, expanded sets of m1 and m2a markers have been. Sebum lipids influence macrophage polarization and activation. Understanding the mysterious m2 macrophage through activation markers and. These markers include transmembrane glycoproteins, scavenger receptors, enzymes, growth factors, hormones, cytokines, and cytokine receptors. Unlike other microglial markers, this marker has the advantage that is isnt expressed by macrophages or other immune or neural cell types.
Axol ipscderived macrophages were harvested at day 7 using cold 1x pbs containing 5 mm edta. Characterization of murine macrophages from bone marrow. American journal of respiratory cell and molecular biology. Novel markers to delineate murine m1 and m2 macrophages. These markers help distinguish macrophages from other similar cell populations and help delineate the macrophages cell of origin, the spatial organization in specific tissues, their unique tissue functions, and the temporal expression of particular markers during macrophages activation in homeostasis and diseases. Macrophage activation marker soluble cd163 associated with. The chemokine system in diverse forms of macrophage activation and polarization. For m1 and m2 macrophages, they have specific markers. Exemplary flow cytometry analysis of day 10 m1macrophages generated in the. Single, live cells were gated, then analysed for the expression of cell surface markers cd45, cd93, and 25f9. Gates for the cell surface markers were drawn using fluorescence minus one fmo controls. Therefore, macrophages from bone marrow, spleen and peritoneum in primary culture are more commonly used. But raw macrophages specifically express higher cd14 than bm macrophage and other cell populations.
No other cell surface markers showed differences for both cell percentages and mfi on small versus large macrophages. Nonparametric tests were used for statistical analysis. A gating strategy for analysis of mature ipscderived macrophages. Flow cytometry characterization of circulating cells belonging to the monocyte macrophage lineage was performed using specific m1 cd80, cd86, tlr2 and tlr4 and m2 surface markers cd204, cd163 and cd206. Cd14 surface molecule is predominantly expressed by cells of myeloid origin and regarded as a specific marker for macrophages m phi. An alveolar macrophage or dust cell is a type of macrophage, a professional phagocyte, found in the pulmonary alveoli, near the pneumocytes, but separated from the wall.
The response of macrophages to endogenous danger signals is only one example of the many different stimuli that trigger macrophage activation in tissues. However, a clear phenotypic characterization of human m1 and m2 subsets is important for a better understanding of their biological functions. A protocol in which thp1 cells were activated with pma then rested in culture pmar more closely resembled. But raw macrophages specifically express higher cd14 than bm macrophage and other. Macrophages remove dead cells in a process named efferocytosis, contributing to the healthy tissue maintenance. Jan, 2010 analysis of cell morphology, cell adhesion, expression of surface markers and phagocytic capacity illustrated that while both vd 3 and pma stimulation induced macrophage differentiation important differences existed in comparison to mdm. Protocol overview using promocell m1m2macrophage generation. Macrophage activation is a complex process involving coordinatesynergistic action of signals from cytokines, chemokines, and pamps. Production and characterization of human macrophages from. Cell surface marker expression with macrophage differentiation. Understanding the mysterious m2 macrophage through activation. Metabolic pathways are closely interconnected and recent literature demonstrates the need for glucose metabolism in antiinflammatory as well as inflammatory macrophages. Activity of the alveolar macrophage is relatively high, because they are located at one of the major boundaries between the body and the outside world.
Additional markers are used to identify specific tissueresident macrophages and different macrophage activation states. The interactive cell markers page shows various cell types and the cell surface markers associated with that cell. There are a large number of commonly used macrophage markers such as cd14, cd16, cd64, cd68, cd71 and ccr5. Exploring the full spectrum of macrophage activation nature. Control of macrophage activation and function by ppars. Cellular energy metabolism is now understood to influence many immune cell pathways as both an indicator and controller of immune cell function and lineage commitment. Exploring the full spectrum of macrophage activation. Exploring the full spectrum of macrophage activation ncbi nih. An example of problematic marker use is expression of arginase1 arg1 as a marker for m2 or mil4 spectrum macrophages, which has led to interpretive problems because arg1 is also induced in m1 spectrum macrophages, expressed in some resident macrophage populations, and highly induced in mycobacteriainfected macrophages, further. Frontiers induction of macrophage function in human thp1. A similar result was found again in the 2 species for the surface markers, lower expression in the pretreated m2 than in the direct m2 macrophage. Macrophage expression of dendritic cell specific icam3 grabbing.
When a b cell in the lymph node recognizes the same unprocessed surface antigen on the bacterium with its surface bound. Are there any specific cell surface markers for m1 and m2 macrophages. Many other listed markers, such as vsig4, gpnmb, cd163 or cd204 can be detected both on the cell surface, and intracellularly. Jci annexin a1 drives macrophage skewing to accelerate. The cytokine environment that is generated by t helper 1 t h 1 or t h 2 cells can have distinct effects on the physiology of macrophages. Macrophages, the sentinels of innate immunity, take residence in nearly every tissue and display marked heterogeneity in their cell surface markers, location, and function.
Additionally, arginase 1 arg1 is a common mouse m2c macrophage marker. Dendritic cell nk cell stem cell precursor macrophage monocyte granulocyte platelet erythrocyte endothelial cell epithelial cell key markers human cd11c cd123 cd56 cd34 hematopoetic stem cell only cd14 cd33 cd66b cd41 cd61 cd62 cd235a cd146 cd326 key markers mouse b cell cd19 cd20 cd45rb220 cd19 cd22 b cell activation marker t cell cd3. Macrophage subpopulation cellsurface expression of immunological markers and phagocytic ability were assessed by flow cytometry. The soluble form of the cd14 molecule scd14, a macrophage activity marker. Where can i find information about the key antibodies to macrophage markers that are available. Today the m2 macrophages are identified based on the expression pattern of a set of m2 markers. Today the m2 macrophages are identified based on the gene transcription or protein expression of a set of m2 markers. For specific macrophage activation the whole volume of the culture is supple. We first characterized the cell surface phenotype of in vitroderived m2 macrophages generated from human monocytes obtained from healthy donors stimulated with mcsf and il4 and compared them to resting macrophages generated with mcsf m0.
Differentiated macrophages are the resident tissue phagocytes and sentinel cells of the innate immune response. A summary of the cell surface marker phenotype of these human monocyte subsets is presented in table 1. We assess recent research in this field, argue for a restricted definition, and explore pathways by which the t helper 2 th2 cell cytokines interleukin4 il4 and il mediate their effects on macrophage cell biology. Flow cytometry for key cell surface markers of innate immunity. The antigen presentation on the surface of infected macrophages in the context of mhc class ii in a lymph node stimulates th1 type 1 helper t cells to proliferate mainly due to il12 secretion from the macrophage. Nonexhaustive list of markers to consider when studying macrophages. Of the 34 surface markers detected on macrophages, 18 altered expression levels on activation. When it comes to investigating the pathways involved in macrophage activation and their pharmacological control, the source of macrophages, the definition of the activators, and the choice of surface markers and transcriptional regulation used to describe the type of activation are critical factors murray et al. The phenotype of mature tissue macrophages represents the composite of environmental and differentiationdependent imprinting. Other markers cell surface, genes or enzymes are easier, but may not track properly with m1kill and m2repair responses best markers are summarized in figure 9 of attached paper, though some. Jun 24, 2017 macrophage surface markers and their capacity to uptake fluorescein isothiocyanate. It promotes macrophage activation, antigen presentation and immune response. Macrophage markers there are a large number of commonly used macrophage markers such as cd14, cd16, cd64, cd68, cd71 and ccr5. When cultured, both human monocyte subsets can differentiate into dcs in the presence of granulocyte macrophage colonystimulating factor gmcsf.
Monocytes and macrophages are phagocytes, acting in both nonspecific defence or innate immunity as well as specific defense or cell mediated immunity of vertebrate animals. Metabolism plays a key role during macrophage activation. What are the t cell, b cell and macrophage activation markers in. However, the contribution of t h 17 cell associated cytokines to macrophage biology is unclear. An evaluation of 2 cohorts of patients with evd revealed that a marker of macrophage activation scd163 but not t cell activation scd25 was associated with severe and fatal evd. Tmem119 is a cell surface protein and a specific microglial marker for both mouse and human. Macrophage activation an overview sciencedirect topics.
Characterisation of lung macrophage subpopulations in copd. Macrophages represent the primary human host response to pathogen infection and link the immediate defense to the adaptive immune system. The process of macrophage activation is described as the polarisation of the. Tissueresident macrophages are a heterogeneous group of cells that express a variety of cell surface and intracellular markers. The concept of an alternative pathway of macrophage activation has stimulated interest in its definition, mechanism, and functional significance in homeostasis and disease. It has been difficult to dissect macrophage phenotypes in vivo, where a spectrum of macrophage phenotypes exists, and also in vitro, where low or nonselective m2 marker protein expression is observed. Learn more about macrophage activation markers including related products, articles. Generally i use cd11c and f480 to id my dendritic cells and cd11b, cd14, f480 to. I think pdl2 is the best cell surface marker for m2a or alternatively activated macrophages, but only for macrophages derived from monocytes. These markers help distinguish macrophages from other similar cell. Displays useful information such as other names, structure, distribution, function, and ligand receptors.
For example cd206 is a good m2 marker in mice but cd200r seems to work better in human cells. Role of tumorassociated macrophages in tumor progression and invasion. I need to know what are the main markers for identifying macrophages and dc from raw 264. It promotes macrophage activation, antigen presentation and immune response 23. Common cell surface markers used to identify human and mouse macrophages include cd11bintegrin alpha m, cd14, cd68, fc gamma riiicd16, fc gamma ricd64, and ccr5, along with f480 in mouse. Il4 andor il produced by t h 2 cells can promote the development of woundhealing macrophages, but these macrophages are poor antigenpresenting cells and may even inhibit t cell. Transcripts encoding chemokines, cytokines, and cell surface were prominent.
High levels of classically activated macrophages with characteristic m1 surface markers localize to visceral fat. C differentially treated macrophages were stained with antibodies against the m2 surface markers cd71 tfr, transferrin receptor, cd206 mrc1, mannose receptor c type 1, cd301 mgl12, macrophage galactosebinding lectin, or isotype control, followed by. Mouse brain tissue sections stained with antitmem119 green ab209064. The agilent advantage measure immune cell activation in minutes rapid, realtime detection of macrophage and t cell activation using agilent seahorse xf technology. Antimouse antibodies against the macrophage markers f480 and cd11b are also included in the panel to positively identify the macrophage. Flow cytometry characterization of circulating cells belonging to the. The alterations in macrophage surface protein expression in response to these stimuli could potentially be used to identify biochemical markers that are unique to these altered cells. No difference was identified when only m1 markers were used. Being these markers for mouse cd71, cd206 and cd301, and cd23, cd200r and cd206 for humans. Mac1 mac3 are surface markers that can be expressed at different percentage levels in different cell population. Furthermore, substantial immunoreactivity of host tissues to a cd163specific antibody, predominantly in areas of extensive immunostaining for ebola virus antigens. Although their markers frequently differ based on their anatomical locations and functions, there is some overlap in the markers that are expressed on different macrophage subpopulations.
Whereas it is admitted that cd163 and cd206 staining labels m2 macrophages, finding m1 immunohistochemistry marker is more challenging. The agilent advantage measure immune cell activation in minutes. Cd163 and cd206 are major markers for the identification of m2 macrophages 16 17 18. Cd47 regulates immune selfrecognition through direct interaction with sirp 2 sirp. Additionally, arginase 1, which is encoded by a signature gene that is induced during alternative macrophage activation, is reduced markedly in both of the ppar. Common macrophage markers models of macrophage activation common macrophage functions. Here, we analyzed the protein kinases of the human monocytic cell line thp1 before and after induction of. Studies have also shown that fizz1, ym1 and ly6c 21 22 can also be used as surface markers associated with m1 or m2 macrophage subpopulations. Monocytes, macrophages, and dendritic cells clinical gate. These markers help distinguish macrophages from other similar cell populations and help delineate the macrophages cell of origin, the spatial organization in specific tissues, their unique tissue functions, and the temporal expression of particular markers during macrophages activation in. However, macrophage activation status within tumors and other settings are.
Biolegend develops and manufactures world class, cuttingedge immunological reagents for biomedical research, offered at an outstanding value. Phorbol12myristateacetate pma and 1,25dihydroxyvitamin d3 vd 3 are stimuli commonly used to induce macrophage differentiation in monocytic cell lines but the extent of differentiation in comparison to primary tissue macrophages is unclear. We examined an array of markers, including mannose and lectin receptors, chemokine receptors, mhcii. Classically m1 and alternatively activated m2 macrophages exhibit distinct phenotypes and functions. Common macrophage markers models of macrophage activation common macrophage functions osteoclasts markers. The alternatively activated or m2 macrophages are immune cells with high phenotypic. M2c macrophages express the cell surface markers, cd163, mmrcd206, and tlr1, and secrete il10 and tgfbeta.
The markers used were cd45 apc fluorophore, immunotools, cd14 pe fluorophore, immunotools, cd11b apc fluorophore, immunotools. Cell surface proteins t, b macrophage, etc flashcards. Rndsylu2945 different serums and in the presence or. A circulating cell population showing both m1 and m2 monocyte. Are there any specific cell surface markers for macrophages. Flow cytometry and highcontent imaging to identify. In macrophage studies, macrophage cell lines including j774a. M2c macrophages, also known as deactivated macrophages, are induced following stimulation with il10, tgfbeta, and glucocorticoids. For example both m1 and m2 will express mhcii, but m1 will express it with much greater intensity. For the results presented in this work, the surface marker panel consists of antibodies against mhc ii, cd86, cd40 and cire cd209.
Graph of median fluorescent intensity mfi of cd14 a and tlr2 b expressed on the cell surface of thp1 cells untreated ut, treated with vitamin d 3 vd 3 or pma treatment with resting pmar and monocytes mo or monocytederived macrophages mdm, n 35, ns p0. Phorbol12myristateacetate pma and 1,25dihydroxyvitamin d3 vd3 are stimuli commonly used to induce macrophage differentiation in monocytic cell. Harvesting murine alveolar macrophages and evaluating. Mature tissue macrophages convert from circulating monocyte precursor cells by terminal differentiation in a process that is not fully understood. Based on a wide flow cytometry surface marker analysis of circulating monocytes macrophages, the study demonstrated that m2 and more significantly cells expressing both m1 and m2 surface markers characterise patients with ssc compared with hss. To provide a foundation for the complexity of in vivo macrophage phenotypes, we performed. Are there any specific cell surface markers for m1 and m2. Similarities and differences in surface receptor expression. Monocyte and macrophage diversity is evidenced by the modulation of cell surface markers and differential production of soluble mediators. These cells are a system because of their location, common origin, similar morphology, shared surface markers, and common functions, particularly phagocytosis. The identification of markers of macrophage differentiation. A growing number of findings highlight the crucial role of metabolic reprogramming in macrophage activation. In obese mice, atms are prone to secrete high levels of tumor necrosis factor. Are there any specific cell surface markers for macrophages and dc both from raw 264.
1132 908 379 574 1029 1338 1372 494 999 321 1187 237 308 1223 670 523 855 737 1472 1493 1275 147 1499 1025 1430 1269 1098 1178 1386 745 465 575 844 836 798 274 1077 342 172 648