Cd14 surface molecule is predominantly expressed by cells of myeloid origin and regarded as a specific marker for macrophages m phi. These markers help distinguish macrophages from other similar cell. However, a clear phenotypic characterization of human m1 and m2 subsets is important for a better understanding of their biological functions. Macrophage subpopulation cellsurface expression of immunological markers and phagocytic ability were assessed by flow cytometry. 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. 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. Learn more about macrophage activation markers including related products, articles. Control of macrophage activation and function by ppars.
A protocol in which thp1 cells were activated with pma then rested in culture pmar more closely resembled. Mac1 mac3 are surface markers that can be expressed at different percentage levels in different cell population. To provide a foundation for the complexity of in vivo macrophage phenotypes, we. Many other listed markers, such as vsig4, gpnmb, cd163 or cd204 can be detected both on the cell surface, and intracellularly. 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. These cells are a system because of their location, common origin, similar morphology, shared surface markers, and common functions, particularly phagocytosis. 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. Cd47 regulates immune selfrecognition through direct interaction with sirp 2 sirp. Today the m2 macrophages are identified based on the expression pattern of a set of m2 markers. Metabolism plays a key role during macrophage activation. Differentiated macrophages are the resident tissue phagocytes and sentinel cells of the innate immune response.
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. 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. Monocytes and tissue macrophages in their various forms see table 1221 on the nelson textbook of pediatrics website at. Generally i use cd11c and f480 to id my dendritic cells and cd11b, cd14, f480 to. Macrophage activation marker soluble cd163 associated with. Monocyte and macrophage diversity is evidenced by the modulation of cell surface markers and differential production of soluble mediators. 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. Classically m1 and alternatively activated m2 macrophages exhibit distinct phenotypes and functions.
For specific macrophage activation the whole volume of the culture is supple. When cultured, both human monocyte subsets can differentiate into dcs in the presence of granulocyte macrophage colonystimulating factor gmcsf. Nonparametric tests were used for statistical analysis. 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. Cd206, the mannose receptor 1, is a ctype lectin mainly present in macrophages and immature dc cells. For example both m1 and m2 will express mhcii, but m1 will express it with much greater intensity. It promotes macrophage activation, antigen presentation and immune response. There are a large number of commonly used macrophage markers such as cd14, cd16, cd64, cd68, cd71 and ccr5.
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. For the results presented in this work, the surface marker panel consists of antibodies against mhc ii, cd86, cd40 and cire cd209. Characterization of murine macrophages from bone marrow. 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. The chemokine system in diverse forms of macrophage activation and polarization. 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 activation an overview sciencedirect topics. 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. 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. A summary of the cell surface marker phenotype of these human monocyte subsets is presented in table 1. When a b cell in the lymph node recognizes the same unprocessed surface antigen on the bacterium with its surface bound.
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. Frontiers induction of macrophage function in human thp1. More recently, expanded sets of m1 and m2a markers have been. Where can i find information about the key antibodies to macrophage markers that are available. Unlike other microglial markers, this marker has the advantage that is isnt expressed by macrophages or other immune or neural cell types. Harvesting murine alveolar macrophages and evaluating. 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. A gating strategy for analysis of mature ipscderived macrophages. Macrophage expression of dendritic cell specific icam3 grabbing. Exploring the full spectrum of macrophage activation nature. The response of macrophages to endogenous danger signals is only one example of the many different stimuli that trigger macrophage activation in tissues. M2c macrophages, also known as deactivated macrophages, are induced following stimulation with il10, tgfbeta, and glucocorticoids.
Mitochondrial dysfunction prevents repolarization of. In obese mice, atms are prone to secrete high levels of tumor necrosis factor. Figure 3 the expression of macrophage markers in small macrophages compared to. Phorbol12myristateacetate pma and 1,25dihydroxyvitamin d3 vd3 are stimuli commonly used to induce macrophage differentiation in monocytic cell. Flow cytometry and highcontent imaging to identify markers. Are there any specific cell surface markers for macrophages. Transcripts encoding chemokines, cytokines, and cell surface were prominent. These markers include transmembrane glycoproteins, scavenger receptors, enzymes, growth factors, hormones, cytokines, and cytokine receptors. Macrophages, the sentinels of innate immunity, take residence in nearly every tissue and display marked heterogeneity in their cell surface markers, location, and function. Jci annexin a1 drives macrophage skewing to accelerate.
The agilent advantage measure immune cell activation in minutes. Similarities and differences in surface receptor expression. 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. What are the t cell, b cell and macrophage activation markers in. Increase in circulating cells coexpressing m1 and m2. Macrophage markers there are a large number of commonly used macrophage markers such as cd14, cd16, cd64, cd68, cd71 and ccr5. Understanding the mysterious m2 macrophage through activation.
Macrophages, by definition, are a type of immune cells capable of engulfing and. Monocytes, macrophages, and dendritic cells clinical gate. Concomitantly, it is also necessary to understand how the in vitro culture conditions used to generate macrophages affect the expression of various surface markers. 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.
Exploring the full spectrum of macrophage activation ncbi nih. Being these markers for mouse cd71, cd206 and cd301, and cd23, cd200r and cd206 for humans. To provide a foundation for the complexity of in vivo macrophage phenotypes, we performed. Macrophages remove dead cells in a process named efferocytosis, contributing to the healthy tissue maintenance. Related surface markers for m2type cells also contain cd68. Common macrophage markers models of macrophage activation common macrophage functions osteoclasts markers. Nonexhaustive list of markers to consider when studying macrophages. Sebum lipids influence macrophage polarization and activation. 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.
The agilent advantage measure immune cell activation in minutes rapid, realtime detection of macrophage and t cell activation using agilent seahorse xf technology. Cell surface marker expression with macrophage differentiation. In macrophage studies, macrophage cell lines including j774a. Cell surface proteins t, b macrophage, etc flashcards. M2c macrophages express the cell surface markers, cd163, mmrcd206, and tlr1, and secrete il10 and tgfbeta. 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.
Are there any specific cell surface markers for m1 and m2 macrophages. Furthermore, substantial immunoreactivity of host tissues to a cd163specific antibody, predominantly in areas of extensive immunostaining for ebola virus antigens. 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. Mature tissue macrophages convert from circulating monocyte precursor cells by terminal differentiation in a process that is not fully understood. Are there any specific cell surface markers for macrophages and dc both from raw 264. There are very few unique macrophage markers and often a number of markers will be required to identify your cell type. 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. These immune cells play key roles in controlling tissue homeostasis, infections, and excessive inflammation. No other cell surface markers showed differences for both cell percentages and mfi on small versus large macrophages. Biolegend develops and manufactures world class, cuttingedge immunological reagents for biomedical research, offered at an outstanding value. Whereas it is admitted that cd163 and cd206 staining labels m2 macrophages, finding m1 immunohistochemistry marker is more challenging.
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. Are there any specific cell surface markers for m1 and m2. Cd163 and cd206 are major markers for the identification of m2 macrophages 16 17 18. Antimouse antibodies against the macrophage markers f480 and cd11b are also included in the panel to positively identify the macrophage. But raw macrophages specifically express higher cd14 than bm macrophage and other cell populations. Axol ipscderived macrophages were harvested at day 7 using cold 1x pbs containing 5 mm edta. Single, live cells were gated, then analysed for the expression of cell surface markers cd45, cd93, and 25f9. Therefore, macrophages from bone marrow, spleen and peritoneum in primary culture are more commonly used. Characterisation of lung macrophage subpopulations in copd. 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. 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. It promotes macrophage activation, antigen presentation and immune response 23. Displays useful information such as other names, structure, distribution, function, and ligand receptors.
The soluble form of the cd14 molecule scd14, a macrophage activity marker. Flow cytometry and highcontent imaging to identify. 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. 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. The phenotype of mature tissue macrophages represents the composite of environmental and differentiationdependent imprinting. Additionally, arginase 1 arg1 is a common mouse m2c macrophage marker. Rndsylu2945 different serums and in the presence or. Of the 34 surface markers detected on macrophages, 18 altered expression levels on activation. High levels of classically activated macrophages with characteristic m1 surface markers localize to visceral fat. Additional markers are used to identify specific tissueresident macrophages and different macrophage activation states. Protocol overview using promocell m1m2macrophage generation.
Macrophages represent the primary human host response to pathogen infection and link the immediate defense to the adaptive immune system. The interactive cell markers page shows various cell types and the cell surface markers associated with that cell. Mouse brain tissue sections stained with antitmem119 green ab209064. Exemplary flow cytometry analysis of day 10 m1macrophages generated in the. A systematic approach to identify markers of distinctly activated. For example cd206 is a good m2 marker in mice but cd200r seems to work better in human cells. I think pdl2 is the best cell surface marker for m2a or alternatively activated macrophages, but only for macrophages derived from monocytes. Jun 24, 2017 macrophage surface markers and their capacity to uptake fluorescein isothiocyanate. Exploring the full spectrum of macrophage activation. From these, expression of 9 surface markers were consistently altered by all conditioning regimens, while 9 were specific to individual polarizing stimuli. Metabolic pathways are closely interconnected and recent literature demonstrates the need for glucose metabolism in antiinflammatory as well as inflammatory macrophages. 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. The alternatively activated or m2 macrophages are immune cells with high phenotypic.
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. The identification of markers of macrophage differentiation. A circulating cell population showing both m1 and m2 monocyte. I need to know what are the main markers for identifying macrophages and dc from raw 264. Production and characterization of human macrophages from. The markers used were cd45 apc fluorophore, immunotools, cd14 pe fluorophore, immunotools, cd11b apc fluorophore, immunotools.
However, the contribution of t h 17 cell associated cytokines to macrophage biology is unclear. Understanding the mysterious m2 macrophage through activation markers and. The concept of an alternative pathway of macrophage activation has stimulated interest in its definition, mechanism, and functional significance in homeostasis and disease. 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. 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. Understanding the mysterious m2 macrophage through. We examined an array of markers, including mannose and lectin receptors, chemokine receptors, mhcii. 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.
Flow cytometry for key cell surface markers of innate immunity. 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 process of macrophage activation is described as the polarisation of the. 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. No difference was identified when only m1 markers were used. 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. 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. Flow cytometry characterization of circulating cells belonging to the. Today the m2 macrophages are identified based on the gene transcription or protein expression of a set of m2 markers. M1 macrophages express specific phenotype markers, including. 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. A growing number of findings highlight the crucial role of metabolic reprogramming in macrophage activation.
Role of tumorassociated macrophages in tumor progression and invasion. 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. American journal of respiratory cell and molecular biology. Plasticity in the macrophage activation states mapping. Moreover, fatty acid oxidation fao not only supports antiinflammatory responses as described formerly but also. Gates for the cell surface markers were drawn using fluorescence minus one fmo controls. Compared with marker cd68, cd163 is more selective to macrophages, so cd163 can be used as a highly specific marker for m2type macrophages 19. Here, we analyzed the protein kinases of the human monocytic cell line thp1 before and after induction of. Tmem119 is a cell surface protein and a specific microglial marker for both mouse and human.
458 78 1441 1329 104 402 1279 118 261 311 1330 1433 1013 948 1074 1429 872 342 950 419 361 1182 1150 1060 620 1409 834 175 979 818 978 49 481 1491 181 1142