Application of an immunocolloid gold technique for the ultrastructural demonstration of IgE-receptor complexes on rat mast cells. 1994

X J Chen, and L Enerbäck
Department of Pathology II, Sahlgrenska Hospital, University of Göteborg, Sweden.

In previous works using cytofluorometry, we demonstrated a broad range of IgE and IgE-receptor levels within individual mast cell populations with a 60 to 80% occupancy of the IgE receptors on mast cells by native IgE. This study was performed in order to confirm our previous findings using an independent method and to visualize the distribution of IgE-receptor complexes on mast cells at an ultrastructural level. For this purpose an indirect immunocolloidal gold-labelling technique has been applied. By counting the number of labelled gold particles, a relative measure of IgE-receptor surface expression and IgE occupancy of the receptors could be obtained. With respect to mast cell morphology and anti-IgE binding specificity criteria, 1% glutaraldehyde + 4% paraformaldehyde (1:1, vol/vol) was found to be the best of the seven fixatives applied in this study. This technique revealed numerous gold particles on the surface of mast cells from barrier-maintained rats (26 +/- 11 per mast cell section, mean +/- SD). Increased numbers of gold particles were counted if the mast cells were incubated with rat myeloma IgE (20 micrograms/ml) (46 +/- 33 per mast cell section, mean +/- SD). There were significantly increased numbers of gold particles on the mast cells of rats infected with N. brasiliensis (126 +/- 30 per mast cell section, mean +/- SD). This indicates that some of the IgE receptors (about 50% of the total number of IgE receptors in this case) on mast cells were occupied by native IgE and that parasite infection significantly increased the number of IgE molecules on the surface of the mast cells. These results correspond with the findings we have made using the cytofluorometric technique and confirm the large individual variations in the density of IgE receptors and IgE among the mast cells of a given cell population. Macrophages, lymphocytes and eosinophils, carrying the low-affinity IgE receptors (Fc epsilon RII), contained less than 5 (normal rats after incubation in rat IgE) or 10 (nematode-infected rats) gold particles per cell section. We also observed some non-granulated lymphocyte-like cells which bound a large number of gold particles after incubation with rat myeloma IgE (20 micrograms/ml), indicating that they contained IgE receptors Fc epsilon RI). They were interpreted as mast cell precursors which have previously been shown to exist in the peritoneal cavity.

UI MeSH Term Description Entries
D007150 Immunohistochemistry Histochemical localization of immunoreactive substances using labeled antibodies as reagents. Immunocytochemistry,Immunogold Techniques,Immunogold-Silver Techniques,Immunohistocytochemistry,Immunolabeling Techniques,Immunogold Technics,Immunogold-Silver Technics,Immunolabeling Technics,Immunogold Silver Technics,Immunogold Silver Techniques,Immunogold Technic,Immunogold Technique,Immunogold-Silver Technic,Immunogold-Silver Technique,Immunolabeling Technic,Immunolabeling Technique,Technic, Immunogold,Technic, Immunogold-Silver,Technic, Immunolabeling,Technics, Immunogold,Technics, Immunogold-Silver,Technics, Immunolabeling,Technique, Immunogold,Technique, Immunogold-Silver,Technique, Immunolabeling,Techniques, Immunogold,Techniques, Immunogold-Silver,Techniques, Immunolabeling
D008297 Male Males
D008407 Mast Cells Granulated cells that are found in almost all tissues, most abundantly in the skin and the gastrointestinal tract. Like the BASOPHILS, mast cells contain large amounts of HISTAMINE and HEPARIN. Unlike basophils, mast cells normally remain in the tissues and do not circulate in the blood. Mast cells, derived from the bone marrow stem cells, are regulated by the STEM CELL FACTOR. Basophils, Tissue,Basophil, Tissue,Cell, Mast,Cells, Mast,Mast Cell,Tissue Basophil,Tissue Basophils
D008854 Microscopy, Electron Microscopy using an electron beam, instead of light, to visualize the sample, thereby allowing much greater magnification. The interactions of ELECTRONS with specimens are used to provide information about the fine structure of that specimen. In TRANSMISSION ELECTRON MICROSCOPY the reactions of the electrons that are transmitted through the specimen are imaged. In SCANNING ELECTRON MICROSCOPY an electron beam falls at a non-normal angle on the specimen and the image is derived from the reactions occurring above the plane of the specimen. Electron Microscopy
D009559 Nippostrongylus A genus of intestinal nematode parasites belonging to the superfamily HELIGMOSOMATOIDEA, which commonly occurs in rats but has been experimentally transmitted to other rodents and rabbits. Infection is usually through the skin. Rat Nematode,Nematode, Rat,Nematodes, Rat,Rat Nematodes
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D016707 Tissue Fixation The technique of using FIXATIVES in the preparation of cytologic, histologic, or pathologic specimens for the purpose of maintaining the existing form and structure of all the constituent elements. Fixation, Tissue
D017206 Strongylida Infections Infections with nematodes of the order STRONGYLIDA. Stephanurus dentatus Infections,Syngamiasis,Angiostrongylus Infections,Infections, Stephanurus dentatus,Infections, Strongylida,Angiostrongylus Infection,Infection, Angiostrongylus,Infection, Stephanurus dentatus,Infection, Strongylida,Stephanurus dentatus Infection,Strongylida Infection,Syngamiases
D017207 Rats, Sprague-Dawley A strain of albino rat used widely for experimental purposes because of its calmness and ease of handling. It was developed by the Sprague-Dawley Animal Company. Holtzman Rat,Rats, Holtzman,Sprague-Dawley Rat,Rats, Sprague Dawley,Holtzman Rats,Rat, Holtzman,Rat, Sprague-Dawley,Sprague Dawley Rat,Sprague Dawley Rats,Sprague-Dawley Rats
D017455 Receptors, IgE Specific molecular sites on the surface of B- and T-lymphocytes which combine with IgEs. Two subclasses exist: low affinity receptors (Fc epsilon RII) and high affinity receptors (Fc epsilon RI). Antigens, CD23,CD23 Antigens,Fc Receptors, epsilon,Fc epsilon Receptor,Fc epsilon Receptors,IgE Receptor,IgE Receptors,Receptors, Fc epsilon,epsilon Fc Receptor,epsilon Fc Receptors,CD 23 Antigens,CD23 Antigen,Fc epsilon RI,Fc epsilon RII,Immunoglobulin E Receptor,Antigen, CD23,Antigens, CD 23,Fc Receptor, epsilon,Receptor, Fc epsilon,Receptor, IgE,Receptor, Immunoglobulin E,Receptor, epsilon Fc,Receptors, epsilon Fc,epsilon RI, Fc,epsilon RII, Fc,epsilon Receptor, Fc,epsilon Receptors, Fc

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