Sep 24, 2018 this is because we wanted to use as much surface markers as possible, including those expressed from mature and polarized macrophages, to better restrict the investigation to monocytes and macrophages, and to study the mixed m1m2 phenotype, independently from the cell maturation state. Frontiers expression of tmem119sall1 and ccr2cd69 in. A protocol in which thp1 cells were activated with pma then rested in culture pmar more closely resembled. Flow cytometry characterization of circulating cells belonging to the.
Enzymatic treatments affect the cell surface expression of cd14 and m2macrophage markers upon polarization, m1 and m2 macrophages express different cell surface molecules which correspond to their ability to perform distinct functions. Cell surface marker an overview sciencedirect topics. American journal of respiratory cell and molecular biology. Are there any phagocytosisassociated surface markers of macrophages. 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. But, surface markers, mostly, are not directly equated with functions. Cd11 also regulates the uptake of complementcoated particles within cells. The exact markers expressed by the monocyte will depend upon the subtype of monocyte and the external stimuli, however monocytes often lack lineage markers for t cells, b cells, nk cells and dc cells, such as. They act not only as part of the first line of defense against invading pathogens, but they are also involved in maintaining tissue integrity and homeostasis. Louis, mo 63110, usa 2department of pathology and immunology, washington university school of medicine, st. Macrophages are located throughout the body and function as professional phagocytic cells that ingest and process foreign materials and cell debris. 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. Although there is a lot of overlap between these cells types, regarding both cell surface markers and function, there are still some distinct features that can. In practice, distinguishing between macrophages and dendritic cells has relied on the use of cell surface markers thought.
In mice, m1 macrophages can be characterized by expression of inos and m2 macrophages by arginase1 production. The role of decidual macrophages during normal and. Single, live cells were gated, then analysed for the expression of cell surface markers cd45, cd93, and 25f9. In addition to the different spectrum of factors released between m1 and m2 states, cell surface markers are altered according to macrophage polarization state. Differential expression of surface markers classifies macrophages to either classically m1 or alternatively activated m2. Phenotype, function, and differentiation potential of. Macrophages are also involved in specific immune responses when recruited by t cells. Humaninduced pluripotent stem cellderived macrophages. M2 macrophages, including m2a, m2b, m2c, m2d subsets, mainly play roles in anti. Modulation of macrophage phenotype by cell shape pnas. Phenotypic and functional characterization of macrophages with. Human alveolar macrophages predominately express combined. 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.
No other cell surface markers showed differences for both cell percentages and mfi on small versus large macrophages. 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 accurate identification and enumeration of these subsets are essential for understanding their role in lung pathology. We investigated whether cd101 may also have a role in the suppressor function of regulatory t cells in humans given that the latter population may affect the autoimmune response in patients with rheumatoid arthritis ra. The precise function of macrophages and their secretory products in ovarian disorders is still. Are there any specific cell surface markers for m1 and m2. Due to local stimuli in their environments or specialized functions associated with. 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.
Cell surface proteins t, b macrophage, etc quizlet. Recently i stained macrophages with f480, cd206 and inos. Production and characterization of human macrophages from. To define macrophage polarization, the field is in urgent need of good surface markers. Recently, the marker cd47 was found to have antiphagocytic signals to. The interactive cell markers page shows various cell types and the cell surface markers associated with that cell. A gating strategy for analysis of mature ipscderived macrophages. Macrophages represent the primary human host response to pathogen infection and link the immediate defense to the adaptive immune system. Each of these macrophage populations can be identified based on their expression or lack of expression of specific cell surface receptors and intracellular markers. Impact of detachment methods on m2 macrophage phenotype.
Characterisation of lung macrophage subpopulations in copd. 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. These macrophages differ in their cell surface markers, secreted cytokines and biological functions. P2y1 function in macrophages is not entirely understood. This macrophage function requires that the t cells release compounds known as lymphokines in response to tumor cells or infected somatic cells. These ipsderived macrophages exhibited the typical morphology of macrophages and were tested for the expression of cell surface markers e. Because normal cells of the body must not be mistakenly removed or compromised, macrophages are selective of. There is gradual up and downregulation of cell surface markers during am ontogeny. Macrophages are specialised cells involved in the detection, phagocytosis and destruction of bacteria and other harmful organisms.
Modulation of macrophage phenotype by cell shape frances y. Humaninduced pluripotent stem cellderived macrophages and. This page covers surface and intracellular cell markers for a variety of cell types including immune cells, stem cells, central nervous system cells, and more. Are there any phagocytosisassociated surface markers of. Some examples include cd14, cd11b, emr1, mac1mac3, lysozyme m, and cd68. In the past, macrophages were classified by the organ in which they were found.
Cd11b, a marker of macrophages and microglia antibody. In practice, distinguishing between macrophages and dendritic cells has relied on the use of cell surface markers. Although there is a lot of overlap between these cells types, regarding both cell surface markers and function, there are still some distinct features that can uniquely identify them. Lymphoid tissue and plasmacytoid dendritic cells and macrophages do not share a common macrophage dendritic cell restricted progenitor. Its an amoebalike organism, and its job is to clean. Structure, receptors, and functions of monocytes and macrophages. Need to define the cell surface markers for macrophage phenotype. The development and function of lungresident macrophages. 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. Related surface markers for m2type cells also contain cd68. The combination of a cd40 agonist with gemcitabine in unresectable pancreatic cancer resulted in regression of tumors by promoting antitumor macrophages.
Cell markers can be expressed both extracellularly on the cells surface or as an intracellular molecule. At the mrna level, we found cd300a, a type i transmembrane protein with a long cytoplasmatic tail, known to regulate immune cells and macrophages 65, 66. Kuppfer cells in the liver, langerhans cells in the skin, microglia in. The role of monocytesmacrophages in hbv and hcv infection. Thus, the cell surface markerbased separation strategies are only an enrichment for mononuclear phagocytes that have functional properties relative to dcs or macrophages for example, antigen. Is it possible for macrophages to express both m1 and m2 markers on the same macrophage. However, distinguishing microglia and monocytederived macrophages histologically can be difficult. Flow cytometric analysis of macrophages and dendritic cell. Macrophages macrophages are located throughout the body and function as professional phagocytic cells that ingest and process foreign materials and cell debris.
Alveolar macrophages am are critical to the homeostasis of the inflammatory environment in the lung. Phenotype, function, and differentiation potential of human. Enzymatic treatments affect the cell surface expression of cd14 and m2macrophage markers. Macrophages function as sentinel cells in the tissues. Abnormalities in macrophage functions have been associated with a wide range of chronic inflammatory and autoimmune diseases. A protocol in which thp1 cells were activated with pma. Macrophages are a diverse group of white blood cells known for eliminating pathogens through phagocytosis. An understanding of the ontogeny, kinetics, and function of phagocytic cells in animals led to the concept of the mononuclear phagocyte system. Surface marker cd163 is upregulated in m2 type macrophage populations, but its upregulation is depending on how you polarized macrophages il4il10. It has recently emerged that most adult tissue macrophages originate during embryonic development and not from circulating monocytes. To examine further how cell elongation influences macrophage phenotype and function, we evaluated arginase1, as well as cd206 macrophage mannose receptor and ym1 by flow cytometry. When cultured, both human monocyte subsets can differentiate into dcs in the presence of granulocyte macrophage colonystimulating factor gmcsf. The mannose receptor, cd206, a prototypic mouse m2 marker, is highly expressed in mcsftreated macrophages in mouse, whereas in human monocytes cd206 is substantially induced by gmcsf 9, 10.
Mature tissue macrophages convert from circulating monocyte precursor cells by terminal differentiation in a process that is not fully understood. A summary of the cell surface marker phenotype of these human monocyte subsets is presented in table 1. M2 macrophages are divided into m2a, m2b, m2c, and m2d subcategories. This notoriously heterogeneous system is composed of macrophages themselves in addition to dendritic cells, monocytes and their lineage committed progenitors. Cell surface proteins t, b macrophage, etc flashcards. Liua,b,c,2 departments of abiomedical engineering and cchemical engineering and materials science, and bthe edwards lifesciences center for advanced cardiovascular technology, university of california, irvine, ca 92697. Gates for the cell surface markers were drawn using fluorescence minus one fmo controls. The methods of cell detachment influence phenotype and function of human macrophages cultured in vitro.
B representative histograms of cell surface markers on monocyte subsets from one of 25 healthy subjects phenotyped c classical, i intermediate, nc nonclassical. 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. We investigated the phenotype of human alveolar macrophages am in adults living in two different geographical locations. Activation and polarization of microglia and macrophages are critical events in neuroinflammation and hematoma resolution after intracerebral hemorrhage ich. What markers can be used to distinguish m1 and m2 macrophages. 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. Documentation of normal function and variation in this complement of subtypes, particularly their differentiation potential to dendritic cells dc or macrophages, remains incomplete. Macrophages are present in all vertebrate tissues, from midgestation throughout life, constituting a widely dispersed organ system. 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. These markers are transmembrane glycoproteins, scavenger receptors, enzymes, growth. Although they share most cell surface markers, evidence indicates that the gene regulation and function of these two cell types might be. Cd11 is involved in numerous adhesionrelated associations between cells such as monocytes, macrophages, natural killer nk cells, and granulocytes. Impact of detachment methods on m2 macrophage phenotype and.
It has also gained usage as a microglial marker for tissues derived from the nervous. I found a large proportion of f480 macrophages are double positive with the expression of both cd206 and inos. The development and function of lungresident macrophages and. Synthesized nanoparticles with il12 payload can reverse macrophages to antitumor function.
Induced pluripotent stem cells ips represent an innovative source for the standardized in vitro generation of macrophages m. Another good example of their tissue specific function would be the removal of apoptotic cells by retinal macrophages, which is key to. In addition, they can also present antigens to t cells and initiate inflammation by releasing molecules known as cytokines that activate other cells. These markers help distinguish macrophages from other similar cell.
Here, we analyzed the protein kinases of the human monocytic cell line thp1 before and after induction of. The cumulative data demonstrating autoimmune aetiologies and abnormal expression of leukocyte cell surface markers may assist in the development of tests which could result in early diagnosis, prior to complete ovarian failure yan et al. Understanding the mysterious m2 macrophage through activation. Kuppfer cells in the liver, langerhans cells in the skin, microglia in the brain and spinal cord, osteoclasts in the bone. Dendritic cells dcs are immune cells and form part of the mammalian immune system.
Displays useful information such as other names, structure, distribution, function, and ligand receptors. Biolegend develops and manufactures world class, cuttingedge immunological reagents for biomedical research, offered at an outstanding value. Protective and pathogenic functions of macrophage subsets. Based on differences in their phenotypes, each of these activated states can be identified based on the expression or lack of expression of certain cell surface and intracellular markers. Macrophages can be polarized into classically activated m1 and alternatively activated m2 macrophages. In practice, distinguishing between macrophages and dendritic cells has relied on the use of cell surface markers thought to be specific to either cell. Mitochondrial dysfunction prevents repolarization of. A circulating cell population showing both m1 and m2 monocyte. They promote homeostasis by responding to internal and external changes within the body, not only as phagocytes in defence against microbes and in clearance of dead and senescent cells, but also through trophic, regulatory and repair functions.
Cd163 and cd206 are major markers for the identification of m2 macrophages 16 17 18. These compounds bind the lymphokine receptors on the macrophages surface, and activate the macrophage to attack nearby cells. But relatively little is known about the consequences of chronic viral infections on monocytes. However, comparative studies defining the influence of cell detachment techniques on secondary characterization of m1 or m2 polarized macrophages are largely absent from the literature. The blood cells that function in allergic reactions and inflammation, contain peroxidase and lysozyme and particularly target parasitic worms and fungi are. The function of macrophages can be divided broadly into two main roles.
Use the left hand navigation to find markers for your cells of interest. Macrophage markers there are a large number of commonly used macrophage markers such as cd14, cd16, cd64, cd68, cd71 and ccr5. Cd40 is a surface marker of macrophages that can be used to inhibit cytotoxic functions. Exploring the full spectrum of macrophage activation. 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 heart is composed of several cell types including cardiomyocytes, cardiac fibroblasts, endothelial cells, smooth muscle cells, and immune cells including cardiac macrophages. The progressive refinement and increasing number of available markers have served to complicate rather than simplify our understanding of the renal mononuclear phagocyte system, and this is.
M1 macrophages express high levels of cd80 and ccr7, whereas m2 macrophages express cd163, cd200r, and cd206 ambarus et al. Receptors located primarily on macrophages and b cells d. Table 2 markers and functions of mc subsets in human and mouse. Therefore, human macrophages from both categories can be characterized by the profile of cytokines produced and the expression of cell surface markers. Just like the related macrophages and dendritic cells, monocyte subtype characterization is complex. In the present study, macrophages derived from hips were.
Cd101 expression and function in normal and rheumatoid. Monocytesmacrophages seem to have a key role in sscrelated ild. The glutamine synthetase glul is a gene associated with glutamine metabolism, an essential pathway for the differentiation and function of macrophages. Macrophages are identified in tissues by their expression of specific protein markers table i, which are predominantly cell surface receptors. Monocytes macrophages constitute the first line of defence for external intrusion or infection. Nov 11, 2019 each of the macrophages has specific protein markers on the cell surface. Induction of macrophage function in human thp1 cells is. In the pages below, you can find guidelines for working with surface molecules to study monocytes. Macrophage contributions to ovarian function human. Cell surface expression of leucocyte markers in freshly isolated and day 7 matured cells was carried out by incubating cells with specific antibodies.
Baines and visser 30 first used the fluorescent dna probe hoescht33342 to separate quiescent cells from the bone marrow. The expression of the surface markers that define mouse m1 and m2 macrophages is different in humans. Circulatory monocytes represent about 10% of leukocytes in human blood and resident macrophages are distributed in a variety of tissues and organs to maintain body homeostasis. Dec 18, 2014 the development and function of lungresident macrophages and dendritic cells. Classical monocytes red, intermediate monocytes yellow, and nonclassical monocytes blue are shown overlaid on negative controls dashed. Recently, the marker cd47 was found to have antiphagocytic signals to macrophages and inhibits natural killer nk cells. After cell detachment, mdm subsets were stained for expression of the canonical macrophage marker cd14, m2macrophage markers cd163, cd200r, cd206, antigen presentation cell markers cd80, cd86, hladr, and fc. The proteins considered most exclusively restricted to macrophages are f480 the human homologue is emr. They act not only as part of the first line of defense against invading pathogens, but they are also involved in. Randolph 2 1center for cardiovascular research, division of cardiology, department of medicine, washington university school of medicine, st. Tissue macrophages are not terminally differentiated and can alter between m1 and m2 phenotypes dependent on the local microenvironment of the cell. Their main function is to process antigen material and present it on their surface to other cells of the immune system. Cell surface markers involved in immune reactions a.
M2 macrophages can be additionally identified by their expression of surface markers, such as mannitol receptor, cd206, cd163, cd209, fizz1, and ym12. However, a clear phenotypic characterization of human m1 and m2 subsets is important for a better understanding of their biological functions. Compared with marker cd68, cd163 is more selective to macrophages, so cd163 can be used as a highly specific marker for m2type macrophages 19. The cd11 protein is actually a heterodimer complex that consists of cd11b and cd18. However, there is no information about human ipsderived hips macrophages hipsm.
Upregulation of cytokines and chemokines, such as il10, tgf. The identification of markers of macrophage differentiation. Mcwhorter a,b,1, tingting wang, phoebe nguyena,b, thanh chunga,b, and wendy f. Macrophage polarization in physiological and pathological. Figure 3 the expression of macrophage markers in small macrophages compared to. A macrophage is a large white blood cell that is an important part of our immune system. The combination of multiple cell surface markers with supravital fluorescent dyes that indicate cell function has become a method of choice for isolating hscs. For m1 and m2 macrophages, they have specific markers.
179 813 613 576 791 429 494 573 27 1154 558 1459 1477 18 372 1471 1002 1538 1597 1310 1000 1388 704 15 1472 919 55 1465 1096 1005 59 328 246 1027 74