5-hydroxytryptamine content of mast cells, mast cell numbers and body growth.
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Fasting gastrinemia, fundic argyrophil cell density, mast cell number, basal fundic histamine content and histidine decarboxylase activity were determined in 20 antrectomized patients and 20 control subjects. Fasting gastrinemia and fundic argyrophil cell density were significantly lower in antrectomized patients than in controls, whereas fundic mast cell number, basal histamine content, and histidine decarboxylase activity did not differ significantly between the two groups. In antrectomized patients the basal fundic histamine content appears related to the fundic mast cell number, as a consequence of the reduced effect of gastrin on argyrophil cells.
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The proliferation of mucosal mast cells (MMC) depends on the presence of interleukin 3 (IL 3) and can be further enhanced by interleukin 4 (IL 4). The supernatant of a TH2 cell clone (ST2/K.9) stimulated by concanavalin A was found to contain a factor, provisionally termed mast cell costimulatory activity (MCA), that substantially enhances the proliferation of MMC promoted by a combination of IL 3 and IL 4. In comparison to other lymphokines MCA is rather resistant to tryptic digestion but is very sensitive to pH values lower than 6.0 and to organic solvents. Chromatographic fractionation of MCA revealed that activity is associated with protein(s) or glycoprotein(s) of 35 to 40 kDa. Partially purified MCA that was functionally free of other T-cell-derived lymphokines did not stimulate mast cell proliferation in the absence of a combination of IL 3 and IL 4. In addition, MCA did not affect the proliferation of mast cells when employed together with either IL 3 or IL 4 alone. Control experiments demonstrated that MCA is identical to neither the T-cell-derived lymphokines IL 2 to IL 6, IL 9, interferon gamma, tumor necrosis factor alpha or beta, or granulocyte-macrophage colony-stimulating factor (CSF), nor to IL 7, granulocyte CSF, macrophage CSF, erythropoietin, leukemia inhibitory factor, or epidermal growth factor (EGF). Finally, experiments using a panel of PPD-reactive TH1- and TH2-like cell lines revealed that MCA is preferentially produced by TH2 cells. These data, especially the relative resistance of MCA to trypsin and the high sensitivity to low pH values and organic solvents, indicate that MCA is distinct from known T-cell-derived lymphokines.(ABSTRACT TRUNCATED AT 250 WORDS)
Mast cells originate from pluripotential cells in the bone marrow. Specifically, human mast cells originate from CD 34-positive progenitor cells. Mast cell proliferation requires IL-3. In the mouse, additional mast cell growth is achieved by the addition of IL-4, and GM-CSF prevents mast cell proliferation. Early bone-marrow-derived mast cells can be identified by their IgE receptors, although they may not yet have the characteristic morphology of mature mast cells. Whether these early cells may by themselves have a physiologic role, remains to be determined. Mast cells persist in culture on fibroblast monolayers, in part due to the production of soluble factor(s) from the fibroblasts themselves. Final mast cell phenotype appears dependent upon the local tissue environment.
A qualitative microspectrophotometric detection method for heparin in situ has been developed, using data obtained previously with a model system of polyacrylamide films containing pure glycosaminoglycans (Tas & Roozemond 1973, and Tas 1975). This technique, based on the unique metachromatic properties of heparin with Toluidine Blue O in glycerol, has been worked out with rat peritoneal and mesenteric mast cells. After equilibration of the stained, air-dried cells in glycerol (for some days), the specific peak of the heparin-Toluidine blue O complex (as found in the model experiments at about 515 nm) could be recorded. It was found that the method can be used to detect unequivocally the presence of heparin in cells, even if they also contain up to 75 mole percent of other, lower sulphated-glycosaminoglycan. Otherwise the method might yield some information about the degree of sulphation of the heparin concerned. With this technique the presence of heparin has been proved in young and adult rat mast cells and for the first time now directly in normal human mast cells and normal human basophilic granulocytes.
Mast cells in adenoid liver tumours of 32 rats induced with nitrosomorpholine were observed ultrastructurally, and among them, some were studied immunocytochemically via immunogold techniques. Data indicated that mast cells which located in tumour tissues presented positive expression of rat mast cell protein (RMCP) II, indicating origination from the mucosal mast cells, while those in the connective tissues around tumours were largely stained negatively with either RMCP II or RMCP I antisera, with the exception of only a few cells showing positive RMCP II staining. Ultrastructural observation showed that mast cells in tumors contacted closely with the tumor cells. Membranes of the intracellular granules in these mast cells were fusing together. The content inside the granules were discharged and spread along the intercellular space between the tumor cells. There was not any lesion obtained ultrastructurally at the contacting point between the tumor cells and the mast cells. The significance of mucosal mast cells in adenoid liver tumors is briefly discussed.
Mast cells are involved in allergic reactions, but may also participate in neurogenic inflammation. The morphology of mast cells in rat dura mater and tongue was evaluated by histochemistry, as well as by scanning and transmission electron microscopy following unilateral trigeminal ganglion stimulation (5 min, 5 Hz, 5 ms, and 0.02, 0.1 or 1.0 mA). Mast cells in dura and tongue of normal animals were numerous, perivascular and often in close proximity to nerve fibers. After 5 min of electrical stimulation, mast cells contralateral to the stimulation showed histochemical characteristics of normal peripheral tissue mast cells (Safranin-positive), and by electron microscopy appeared homogeneous with numerous intact electron-dense granules. On the stimulated side, however, the staining characteristics of mast cells showed changes indicating progressive intracellular loss of their granular content. In addition, the total number of stainable mast cells decreased at all three stimulus intensities, but reached significance only at 0.1 and 0.02 mA. Ultrastructural evidence of granule changes consistent with secretion were observed although degranulation was not observed until 20 min after stimulation. There were no mast cell changes after electrical trigeminal stimulation in adult rats treated as neonates with capsaicin to destroy small caliber sensory afferent axons. These results suggest that mast cells may secrete in response to electrical stimulation of trigeminal axons, possibly mediated by antidromic release of neuropeptides, and may participate in the development of neurogenic inflammation.
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Mast cells purified from the peritoneal cell population and mast cells derived in culture from bone marrow cells were examined for their sensitivity to murine cytomegalovirus (MCMV) infection in vitro. While up to 70% of mast cells expressed viral antigens, less than 12% of the cells produced infectious virus. Transmission electron microscopy demonstrated nucleocapsids in the nuclei and in association with the cisternal elements of the Golgi apparatus. Some complete virions were found within small cytoplasmic vacuoles. In contrast with previous studies of macrophages and fibroblasts, the susceptibility of mast cells to MCMV infection in vitro was not influenced by the H-2 or non-H-2 genotype of the donor.
Mast cells isolated from feline splenic mastocytomas were cultured to study their structural and functional properties. Isolated cells from various cats were grown as monolayer cultures for a mean of 56 days (range, 30 to 76 days). Cat mast cells released allergic mediators in response to compound 48/80, anti-cat serum antibodies, and concanavalin A. On the basis of the finding that secretion from cat mast cells was stimulated by anti-cat serum antibodies and concanavalin A, these cells contain surface-bound immunoglobulins. The presence of mast cell-sensitizing antibodies has been suspected in cats, but never before directly demonstrated. Cultured cat mast cells have cytochemical and functional characteristics common to connective tissue-type mast cells and provide one of the few non-rodent models of cultured cells for the study of this type of mast cell.
Mast cells and basophils, although sharing many constitutive properties, are quite distinct in their development, functions and biological properties. Mast cell granules are composed of a macromolecular matrix of proteoglycan and neutral protease of which heparin and tryptase, respectively, are predominant. The distribution of the other major neutral protease, chymase, allows human mast cell subpopulations to be subdivided immunocytochemically. All human mast cells respond to IgE-dependent stimulation with the secretion of the preformed mediator, histamine, and the newly generated lipid-derived eicosanoids PGD2 and LTC4. Although amounts of these products vary between mast cells dispersed from different tissues, it is uncertain whether this reflects true heterogeneity. Mast cells of the human skin, but not those of other tissues, are sensitive to stimulation by substance P, compound 48/80 and other basic non-immunological stimuli. The mechanism of mediator secretion induced by these agents is distinct from that induced by IgE-dependent stimulation. However, the morphological characteristics of degranulation are similar, suggesting that the distinct biochemical pathways merge into a common pathway before effecting degranulation.
Mast cell counts were made, in tissue samples of normal and clubbed fingers obtained at autopsy, to investigate the possibility that mast cells act as an intermediary in the development of finger clubbing. Quantitative measurements of the composition of the tissues of the normal and clubbed nail bed were also made. The only significant difference in mast cell numbers in clubbed fingers was a decrease in mast cells in the tissues just superficial to the nail. The appearance of the mast cells was normal. The porportion of blood vessels and other tissues was not significantly different from normal.
Mast cells have characteristic granulae containing various glucoseaminoglycans, proteases and amines (predominantly histamine). The conventional histological methods for studying mast cells are based upon acidic ortho- and metachromatic routine stains of the glucoseaminoglycans. However, the success of these procedures is dependent upon both the fixatives and the tissues used. In this study, we wanted to find out whether an immunohistochemical procedure could overcome some of these difficulties. Normal human skin was fixed in five different types of fixative and processed for indirect immunofluorescence, using an antiserum to histamine. Only one, 4% carbodiimide in 0.1 M phosphate buffer (pH 7.4), resulted in immunostaining. The quality of the staining was good, with a high signal-to-noise ratio, and was located on the mast cells. The method made it possible to visualize small structures such as a single secreted granula, the thin cytoplasmatic extension of some cells, and a previously undescribed dendritic morphology of some of the mast cells. We therefore recommend this procedure for cellular studies of mast cells when accuracy is needed.
Mast cells are present in the arterial intima, the site of atherogenesis. To gain insight into the possible role of mast cells in the formation of the cholesterol-loaded macrophage foam cells typical of both early and late atherosclerotic lesions, a model system was developed in which isolated rat serosal mast cells were incubated with mouse peritoneal macrophages in medium to which low-density lipoproteins (LDL) had been added. Stimulation of the mast cells was found to induce a 50-fold enhancement of LDL uptake by the macrophages, which concomitantly accumulated LDL-derived cholesterol. This process, called the "granule-mediated uptake of LDL", involves the following steps: (i) exocytosis of the cytoplasmic granules of the mast cells, (ii) escape of soluble granule components, such as histamine and a fraction of the granule heparin proteoglycans into the medium, leaving granule remnants consisting of neutral proteases embedded in a heparin proteoglycan matrix, (ii) binding of LDL to binding sites on the glycosaminoglycan side chains of the heparin proteoglycan component of the granule remnants, (iv) proteolytic degradation of the bound LDL by the neutral proteases of the granule remnants, (v) fusion of degraded LDL particles on the surfaces of the granule remnants, and (vi) phagocytosis of the LDL-laden granule remnants by the macrophages. Simultaneously, the soluble heparin proteoglycans, to which no proteolytic enzymes are bound, interact with LDL with formation of insoluble complexes which are also phagocytosed by the macrophages.(ABSTRACT TRUNCATED AT 250 WORDS)
Mast cells isolated from rat peritoneal and pleural cavities were incubated in vitro with radioactively-labelled histamine (Hi), 5-hydroxytryptamine (5-HT), dopamine (DA), noradrenaline (NA), tyramine (TA), phenylethylamine (PhEA), trptamine (TrpA), ephedrine (Eph) or amphetamine (Amph). All these amines were taken up by the mast cells. The dose-response curves for the compound 48/80-induced release of endogenous Hi and for the various amines taken up by the cells were compared. The release curves for Hi, 5-HT, DA, NA and TA were found to be similar to that for endogenous Hi, while those for PhEA, TrpA, Eph and Amph were different from that for endogenous Hi. The uptake of Hi, 5-HT, DA, PhEA, TrpA and Eph into granules in mast cells was studied. Membrane-bound granules were obtained by sonication of mast cells incubated with the respective amine, followed by differential contrifugation. The amine content of these granules was then measured. Hi, 5-HT and DA were found to be mainly localized to the granules, while a smaller proportion of the PhEA, TrpA and Eph was found there, the rest being located extragranularly. The present results suggest that, when taken up by rat mast cells, even amines which are not endogenous to the cells are stored in the same way as the endogenous amines Hi and 5-HT.
The cDNA and gene for mouse mast cell protease-6 (MMCP-6) have been sequenced and show MMCP-6 to be translated as a prepro-enzyme with a 21-amino acid hydrophobic leader peptide, a 10-amino acid activation peptide, and a 245-amino acid mature enzyme. The mature form of the enzyme has 73% amino acid sequence identity with human and dog mast cell tryptases. The MMCP-6 gene includes 6 exons, with a total span of 1.8 kilobases. A 208-base pair intron was defined which separates the 5'-untranslated sequence of MMCP-6 from the translation initiation codon, thereby presenting a gene organization which distinguishes tryptic serine proteases from chymotryptic serine proteases of the mast cell secretory granule. By RNA blot analysis with a gene-specific probe, MMCP-6 has a unique subclass distribution in being transcribed in mouse connective tissue mast cells but undetectable in mucosal mast cells. MMCP-6 is the first serine protease of any class to be shown to be significantly transcribed in progenitor, bone marrow-derived mast cells, which can reconstitute both mucosal mast cell and connective tissue mast cell populations in mast cell-deficient mice.