Purification and characterization of human skin mast cells. Evidence for human mast cell heterogeneity
Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, Good Samaritan Hospital 21239. The Journal of Immunology
(Impact Factor: 4.92).
Our previous studies of human lung and intestinal mast cells failed to show the heterogeneity found among mast cells in murine species. Recently, we and others have developed techniques for the enzymatic dispersion of human neonatal skin mast cells. In addition, we are now able to make single cell suspensions of mast cells from adult skin and to purify these cells to near homogeneity. Comparative studies of mast cells from these several sources have uncovered several major differences among them. Adult and neonatal skin mast cells themselves differ in that the former are 10-fold less sensitive to goat anti-human IgE, with maximal release occurring at 3.0 and 0.3 microgram/ml, respectively. Skin mast cells also differ in optimal temperature for release: adult mast cells respond maximally at 23 to 30 degrees C and neonatal cells at 37 degrees C. Skin mast cells from both sources are dramatically different from lung and intestinal mast cells in two aspects. First, skin mast cells are quite responsive to several stimuli--morphine sulfate (10(-4) to 10(-6) M), substance P (10(-5) to 10(-7) M), compound 48/80 (10 to 0.1 microgram/ml), f-Met peptide (10(-6) M), and C5a (10(-8) M)--to which the other mast cells fail to respond. Second, although stimulated skin mast cells produce prostaglandin D2, little leikotriene C4, if any, is generated, unlike lung or intestinal mast cells. These differences in inflammatory potential among human mast cells from various sites have important implications for the management of allergic and inflammatory responses.
Available from: Domenico Ribatti
- "Some difference in the granule structure was recognized. Lung, skin and synovial mast cells were detected to contain approximately 4 pg of histamine per cell, whereas the amount of histamine in basophils was about 1 pg per cell   . Human lung mast cells contained a mixture of both heparin and chondroitin sulphate E, whereas chondroitin sulphate A was recognized as the dominant proteoglycan in basophil granules  . "
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ABSTRACT: Work on mast cells and basophils began with their identification by Paul Ehrlich at the end of the 19th century. Mast cells and basophils were immediately perceived as closely linked cells and early nomenclature formulated by Ehrlich himself, i.e., tissue "Mastzellen" and blood "Mastzellen", reflected this unifying viewpoint. With time, important functional affinities but also substantial diversities were recognized. This review article focuses on the historical development of the concept of mast cell/basophil specificity, from the initial identification of these cells to current studies.
Available from: Peter N Monk
- "Thus, it appears that subsets of lymphocytes may be responsive to C5a and this percentage increases following activation. Mast cells also show highly variable responsiveness to C5a; for instance, skin mast cells respond to C5a, whereas lung and intestinal mast cells do not (Lawrence et al., 1987). More recently, expression of a receptor for C5a has been shown to discriminate between mast cell subsets, which also show distinct differences in protease expression, suggesting that C5a responsiveness is programmed into mast cell development (Oskeritzian et al., 2005). "
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ABSTRACT: Complement fragment (C)5a is a 74 residue pro-inflammatory polypeptide produced during activation of the complement cascade of serum proteins in response to foreign surfaces such as microorganisms and tissue damaged by physical or chemical injury. C5a binds to at least two seven-transmembrane domain receptors, C5aR (C5R1, CD88) and C5L2 (gpr77), expressed ubiquitously on a wide variety of cells but particularly on the surface of immune cells like macrophages, neutrophils and T cells. C5aR is a classical G protein-coupled receptor that signals through Gαi and Gα16, whereas C5L2 does not appear to couple to G proteins and has no known signalling activity. Although C5a was first described as an anaphylatoxin and later as a leukocyte chemoattractant, the widespread expression of C5aR suggested more general functionality. Our understanding of the physiology of C5a has improved significantly in recent years through exploitation of receptor knockout and knockin mice, C5 and C5a antibodies, soluble recombinant C5a and C5a analogues and newly developed receptor antagonists. C5a is now also implicated in non-immunological functions associated with developmental biology, CNS development and neurodegeneration, tissue regeneration, and haematopoiesis. Combined receptor mutagenesis, molecular modelling, structure-activity relationship studies and species dependence for ligand potency on C5aR have been helpful for identifying ligand binding sites on the receptor and for defining mechanisms of receptor activation and inactivation. This review will highlight major developments in C5a receptor research that support C5aR as an important therapeutic target. The intriguing possibilities raised by the existence of a non-signalling C5a receptor are also discussed.
British Journal of Pharmacology (2007) 152, 429–448; doi:10.1038/sj.bjp.0707332; published online 2 July 2007
Available from: K. Frank Austen
- "hMCs differ in their profiles of eicosanoid biosynthesis in response to FcεRI-dependent activation after isolation from various dispersed tissue sources. Both total cys-LT generation (from 3.5 ng/106 hMCs from skin to 45 ng/106 hMCs from uterus) and the cys-LT/PGD2 ratio (1:12 for skin hMCs; 1:3, 1:2, and 1:1 for lung, uterine, and intestinal hMCs, respectively) are marked by tissue-related differences that are both quantitative and relative 31323334. Because MCs in all tissues derive from a single lineage of circulating committed progenitors 23536 under the influence of constitutively expressed SCF, we postulated that their heterogeneous profiles of eicosanoid generation would be determined by the absence or presence of additional local factors, particularly the cytokines derived from the Th2 lymphocytes that associate with mucosal surfaces in allergic diseases. "
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ABSTRACT: Human mast cells (hMCs) derived in vitro from cord blood mononuclear cells exhibit stem cell factor (SCF)-dependent comitogenic responses to T helper cell type 2 (Th2) cytokines. As cysteinyl leukotriene (cys-LT) biosynthesis is a characteristic of immunoglobulin (Ig)E-activated mucosal hMCs, we speculated that Th2 cytokines might regulate eicosanoid generation by hMCs. After passive sensitization for 5 d with IgE in the presence of SCF, anti-IgE-stimulated hMCs elaborated minimal cys-LT (0.1 +/- 0.1 ng/10(6) hMCs) and abundant prostaglandin (PG)D(2) (16.2 +/- 10.3 ng/10(6) hMCs). Priming of hMCs by interleukin (IL)-4 with SCF during passive sensitization enhanced their anti-IgE-dependent histamine exocytosis and increased their generation of both cys-LT (by 27-fold) and PGD(2) (by 2. 5-fold). Although priming with IL-3 or IL-5 alone for 5 d with SCF minimally enhanced anti-IgE-mediated cys-LT generation, these cytokines induced further six- and fourfold increases, respectively, in IgE-dependent cys-LT generation when provided with IL-4 and SCF; this occurred without changes in PGD(2) generation or histamine exocytosis relative to hMCs primed with IL-4 alone. None of these cytokines, either alone or in combination, substantially altered the levels of cytosolic phospholipase A(2) (cPLA(2)), 5-lipoxygenase (5-LO), or 5-LO activating protein (FLAP) protein expression by hMCs. In contrast, IL-4 priming dramatically induced the steady-state expression of leukotriene C(4) synthase (LTC(4)S) mRNA within 6 h, and increased the expression of LTC(4)S protein and functional activity in a dose- and time-dependent manner, with plateaus at 10 ng/ml and 5 d, respectively. Priming by either IL-3 or IL-5, with or without IL-4, supported the localization of 5-LO to the nucleus of hMCs. Thus, different Th2-derived cytokines target distinct steps in the 5-LO/LTC(4)S biosynthetic pathway (induction of LTC(4)S expression and nuclear import of 5-LO, respectively), each of which is necessary for a full integrated functional response to IgE-dependent activation, thus modulating the effector phenotype of mature hMCs.
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