Are there any specific cell surface markers for m1 and m2 macrophages. 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. Axol ipscderived macrophages were harvested at day 7 using cold 1x pbs containing 5 mm edta. M2c macrophages express the cell surface markers, cd163, mmrcd206, and tlr1, and secrete il10 and tgfbeta. Biolegend develops and manufactures world class, cuttingedge immunological reagents for biomedical research, offered at an outstanding value. For example cd206 is a good m2 marker in mice but cd200r seems to work better in human cells. Mac1 mac3 are surface markers that can be expressed at different percentage levels in different cell population. These markers are transmembrane glycoproteins, scavenger receptors, enzymes, growth. High levels of classically activated macrophages with characteristic m1 surface markers localize to visceral fat. Protocol overview using promocell m1m2macrophage generation. 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. Macrophage markers there are a large number of commonly used macrophage markers such as cd14, cd16, cd64, cd68, cd71 and ccr5. 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. Cell surface proteins t, b macrophage, etc flashcards.
But raw macrophages specifically express higher cd14 than bm macrophage and other. Generally i use cd11c and f480 to id my dendritic cells and cd11b, cd14, f480 to. The response of macrophages to endogenous danger signals is only one example of the many different stimuli that trigger macrophage activation in tissues. Increase in circulating cells coexpressing m1 and m2. 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. The markers used were cd45 apc fluorophore, immunotools, cd14 pe fluorophore, immunotools, cd11b apc fluorophore, immunotools. When cultured, both human monocyte subsets can differentiate into dcs in the presence of granulocyte macrophage colonystimulating factor gmcsf. Tissueresident macrophages are a heterogeneous group of cells that express a variety of cell surface and intracellular markers. 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. Today the m2 macrophages are identified based on the gene transcription or protein expression of a set of m2 markers.
Exploring the full spectrum of macrophage activation nature. Phorbol12myristateacetate pma and 1,25dihydroxyvitamin d3 vd3 are stimuli commonly used to induce macrophage differentiation in monocytic cell. When a b cell in the lymph node recognizes the same unprocessed surface antigen on the bacterium with its surface bound. Unlike other microglial markers, this marker has the advantage that is isnt expressed by macrophages or other immune or neural cell types.
Jci annexin a1 drives macrophage skewing to accelerate. What are the t cell, b cell and macrophage activation markers in. Additionally, arginase 1 arg1 is a common mouse m2c macrophage marker. Cd206, the mannose receptor 1, is a ctype lectin mainly present in macrophages and immature dc cells. Additional markers are used to identify specific tissueresident macrophages and different macrophage activation states. Understanding the mysterious m2 macrophage through. These markers help distinguish macrophages from other similar cell.
The soluble form of the cd14 molecule scd14, a macrophage activity marker. 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. Characterisation of lung macrophage subpopulations in copd. 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.
Flow cytometry for key cell surface markers of innate immunity. Moreover, fatty acid oxidation fao not only supports antiinflammatory responses as described formerly but also. Whereas it is admitted that cd163 and cd206 staining labels m2 macrophages, finding m1 immunohistochemistry marker is more challenging. 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. From these, expression of 9 surface markers were consistently altered by all conditioning regimens, while 9 were specific to individual polarizing stimuli. The chemokine system in diverse forms of macrophage activation and polarization. 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, the contribution of t h 17 cell associated cytokines to macrophage biology is unclear. The process of macrophage activation is described as the polarisation of the. Exploring the full spectrum of macrophage activation. 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. M1 macrophages express specific phenotype markers, including.
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. No difference was identified when only m1 markers were used. Flow cytometry and highcontent imaging to identify markers. Of the 34 surface markers detected on macrophages, 18 altered expression levels on activation. There are very few unique macrophage markers and often a number of markers will be required to identify your cell type. Metabolic pathways are closely interconnected and recent literature demonstrates the need for glucose metabolism in antiinflammatory as well as inflammatory macrophages. 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. Exploring the full spectrum of macrophage activation ncbi nih. Macrophages, the sentinels of innate immunity, take residence in nearly every tissue and display marked heterogeneity in their cell surface markers, location, and function. Learn more about macrophage activation markers including related products, articles. 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. Are there any specific cell surface markers for macrophages.
A growing number of findings highlight the crucial role of metabolic reprogramming in macrophage activation. The concept of an alternative pathway of macrophage activation has stimulated interest in its definition, mechanism, and functional significance in homeostasis and disease. Macrophage activation marker soluble cd163 associated with. Understanding the mysterious m2 macrophage through activation. 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. Gates for the cell surface markers were drawn using fluorescence minus one fmo controls. For the results presented in this work, the surface marker panel consists of antibodies against mhc ii, cd86, cd40 and cire cd209. Plasticity in the macrophage activation states mapping. Nonexhaustive list of markers to consider when studying macrophages. 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. 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. Being these markers for mouse cd71, cd206 and cd301, and cd23, cd200r and cd206 for humans.
It promotes macrophage activation, antigen presentation and immune response 23. A circulating cell population showing both m1 and m2 monocyte. 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. 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. No other cell surface markers showed differences for both cell percentages and mfi on small versus large macrophages.
Macrophage activation is a complex process involving coordinatesynergistic action of signals from cytokines, chemokines, and pamps. Exemplary flow cytometry analysis of day 10 m1macrophages generated in the. Macrophage expression of dendritic cell specific icam3 grabbing. Transcripts encoding chemokines, cytokines, and cell surface were prominent. Differentiated macrophages are the resident tissue phagocytes and sentinel cells of the innate immune response. It promotes macrophage activation, antigen presentation and immune response. Frontiers induction of macrophage function in human thp1. Sebum lipids influence macrophage polarization and activation. 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. The alternatively activated or m2 macrophages are immune cells with high phenotypic. I need to know what are the main markers for identifying macrophages and dc from raw 264. Monocytes, macrophages, and dendritic cells clinical gate. The phenotype of mature tissue macrophages represents the composite of environmental and differentiationdependent imprinting. 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.
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. 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. Displays useful information such as other names, structure, distribution, function, and ligand receptors. Related surface markers for m2type cells also contain cd68. Jun 24, 2017 macrophage surface markers and their capacity to uptake fluorescein isothiocyanate. These cells are a system because of their location, common origin, similar morphology, shared surface markers, and common functions, particularly phagocytosis.
The agilent advantage measure immune cell activation in minutes rapid, realtime detection of macrophage and t cell activation using agilent seahorse xf technology. 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. Macrophage activation an overview sciencedirect topics. Furthermore, substantial immunoreactivity of host tissues to a cd163specific antibody, predominantly in areas of extensive immunostaining for ebola virus antigens. These immune cells play key roles in controlling tissue homeostasis, infections, and excessive inflammation. Common macrophage markers models of macrophage activation common macrophage functions. Cell surface marker expression with macrophage differentiation.
Macrophages, by definition, are a type of immune cells capable of engulfing and. Mouse brain tissue sections stained with antitmem119 green ab209064. Flow cytometry characterization of circulating cells belonging to the. Flow cytometry and highcontent imaging to identify. Therefore, macrophages from bone marrow, spleen and peritoneum in primary culture are more commonly used. Are there any specific cell surface markers for m1 and m2. Monocyte and macrophage diversity is evidenced by the modulation of cell surface markers and differential production of soluble mediators.
Mitochondrial dysfunction prevents repolarization of. Antimouse antibodies against the macrophage markers f480 and cd11b are also included in the panel to positively identify the macrophage. 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. Concomitantly, it is also necessary to understand how the in vitro culture conditions used to generate macrophages affect the expression of various surface markers. Tmem119 is a cell surface protein and a specific microglial marker for both mouse and human. To provide a foundation for the complexity of in vivo macrophage phenotypes, we performed. In obese mice, atms are prone to secrete high levels of tumor necrosis factor. A gating strategy for analysis of mature ipscderived macrophages.
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. Cd163 and cd206 are major markers for the identification of m2 macrophages 16 17 18. However, macrophage activation status within tumors and other settings are. Characterization of murine macrophages from bone marrow. 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. Figure 3 the expression of macrophage markers in small macrophages compared to. For example both m1 and m2 will express mhcii, but m1 will express it with much greater intensity. A protocol in which thp1 cells were activated with pma then rested in culture pmar more closely resembled. More recently, expanded sets of m1 and m2a markers have been. The interactive cell markers page shows various cell types and the cell surface markers associated with that cell. Mature tissue macrophages convert from circulating monocyte precursor cells by terminal differentiation in a process that is not fully understood.
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. 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. To provide a foundation for the complexity of in vivo macrophage phenotypes, we. Here, we analyzed the protein kinases of the human monocytic cell line thp1 before and after induction of. However, a clear phenotypic characterization of human m1 and m2 subsets is important for a better understanding of their biological functions. Single, live cells were gated, then analysed for the expression of cell surface markers cd45, cd93, and 25f9. Cd14 surface molecule is predominantly expressed by cells of myeloid origin and regarded as a specific marker for macrophages m phi. Monocytes and macrophages are phagocytes, acting in both nonspecific defence or innate immunity as well as specific defense or cell mediated immunity of vertebrate. 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.
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. A systematic approach to identify markers of distinctly activated. Nonparametric tests were used for statistical analysis. Novel markers to delineate murine m1 and m2 macrophages. In macrophage studies, macrophage cell lines including j774a. The identification of markers of macrophage differentiation. Similarities and differences in surface receptor expression. Cd47 regulates immune selfrecognition through direct interaction with sirp 2 sirp. 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. Macrophages represent the primary human host response to pathogen infection and link the immediate defense to the adaptive immune system. We examined an array of markers, including mannose and lectin receptors, chemokine receptors, mhcii. Rndsylu2945 different serums and in the presence or.
There are a large number of commonly used macrophage markers such as cd14, cd16, cd64, cd68, cd71 and ccr5. But raw macrophages specifically express higher cd14 than bm macrophage and other cell populations. Understanding the mysterious m2 macrophage through activation markers and. Metabolism plays a key role during macrophage activation. Monocytes and tissue macrophages in their various forms see table 1221 on the nelson textbook of pediatrics website at. Role of tumorassociated macrophages in tumor progression and invasion. 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. Control of macrophage activation and function by ppars.
Macrophages remove dead cells in a process named efferocytosis, contributing to the healthy tissue maintenance. Are there any specific cell surface markers for macrophages and dc both from raw 264. A summary of the cell surface marker phenotype of these human monocyte subsets is presented in table 1. Elevated proinflammatory cytokine levels and markers of m1 macrophages are found in ppar. For m1 and m2 macrophages, they have specific markers. The agilent advantage measure immune cell activation in minutes. 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. Macrophage subpopulation cellsurface expression of immunological markers and phagocytic ability were assessed by flow cytometry. Compared with marker cd68, cd163 is more selective to macrophages, so cd163 can be used as a highly specific marker for m2type macrophages 19. American journal of respiratory cell and molecular biology. 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. M2c macrophages, also known as deactivated macrophages, are induced following stimulation with il10, tgfbeta, and glucocorticoids.
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 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. 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. Many other listed markers, such as vsig4, gpnmb, cd163 or cd204 can be detected both on the cell surface, and intracellularly.
Classically m1 and alternatively activated m2 macrophages exhibit distinct phenotypes and functions. These markers include transmembrane glycoproteins, scavenger receptors, enzymes, growth factors, hormones, cytokines, and cytokine receptors. Common macrophage markers models of macrophage activation common macrophage functions osteoclasts markers. Production and characterization of human macrophages from.
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