Advances in Anatomy Embryology and Cell Biology Vol. 114 Editors F. Beck, Leicester W Hild, Galveston W Kriz, Heidelberg R. Ortmann, Kaln J.E. Pauly, Little Rock T.H. Schiebler, Wiirzburg Ann M. Dvorak Human Mast Cells With 89 Figures Springer-Verlag Berlin Heidelberg New York London Paris Tokyo Dr. Ann M. Dvorak Department of Pathology, Beth Israel Hospital 330 Brookline Avenue, Boston, MA 02215, USA ISBN-13: 978-3-540-50374-3 e-ISBN-13: 978-3-642-74145-6 DOl: 10.1007/978-3-642-74145-6 Library of Congress Cataloging-in-Publication Data Dvorak. Ann M. Human mast cells/Ann M. Dvorak. p. cm.-(Advances in anatomy, embryology, and cell biology; vol. 114) Bibliography: p. ISBN-13: 978-3-540-50374-3 (U.S.) 1. Mast cells-Immunology. 2. Anaphylaxis-Pathogenesis. I. Title. II. Series: Advances in anatomy, embryology, and cell biology; v. 114. QL80l.E67 vol. 114 [QRI85.8.M35] 574.4 s [616.07'9] 88-31849 This work is subject to copyright. 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Typesetting: Macmillan India Ltd, Bangalore-25, India 2121/3140-54321O-Printed on acid-free paper For my family H aroid, John, Laurie, and Jane Contents 1 Introduction 2 Ultrastructure of Human Mast Cells . 3 3 Ultrastructure of Human Basophils . 13 4 In Vitro Studies of Human Mast Cells 15 4.1 Human Mast Cell Granules and Lipid Bodies 15 4.1.1 Differences in Structure . . . . . . 15 4.1.2 Differences in Content. . . . . . . . . 15 4.1.3 Differences in Mechanism of Formation 21 4.1.4 Differences in Behavior During Degranulation 22 4.1.5 Differences in Behavior During Recovery from Degranulation. . . . . . . . . . . . 25 4.2 Anaphylactic Degranulation of Human Lung Mast Cells In Vitro. . . . . . . . . . . . .. 27 4.3 Recovery from Anaphylactic Degranulation of Human Lung Mast Cells In Vitro. 48 4.3.1 Early Recovery Events. . 48 4.3.2 Late Recovery Events. . 65 4.3.3 Mixed Recovery Patterns. 72 5 In Vivo Studies of Human Mast Cells 74 5.1 Anaphylactic Degranulation of Human Mast Cells In Vivo. . . . . . . . . . . . . " 74 5.2 Piecemeal Degranulation of Human Mast Cells In Vivo. . . . . . . . . . . . . .. 75 5.3 Activated Human Mast Cells Have Increased Lipid. Bodies . . . . . 81 5.4 Mast Cell Shedding. . . . 82 6 Mast Cell Morphologic Cycles 83 6.1 In Vitro . 83 6.2 In Vivo. . . . . . . . . 88 VII 7 Concluding Remarks 89 8 Summary 92 References . 95 Subject Index 101 VIII 1 Introduction Mast cells and basophils were first described by Ehrlich (1877, 1878, 1879). Although these cells share many functional properties, they can readily be distinguished using morphological criteria (Dvorak 1986a; Dvorak et al. 1983a, 1983c; Galli et al. 1984). The identification of immunoglobulin E (IgE) and high affinity IgE receptors on mast cells and basophils was instrumental to our understanding of the mechanisms underlying the role of these cells in immediate hypersensitivity reactions (Ishizaka and Ishizaka 1979; Ishizaka et al. 1966, 1972, 1973; Tomioka and Ishizaka 1971). We now know that these IgE-mediated mechanisms as well as a number of other stimuli can cause the rapid release of many preformed mediators of inflammation from both mast cells and basophils (Galli et al. 1984). The most well-known of these is histamine. Potent mediators that are not preformed are also stimulated and released from these cells. Recently, products of arachidonic acid metabolism, such as the prostaglandins and leukotrienes, have been found to be generated either by the cyclooxygenase pathway or the lipoxy genase pathway in mast cells and basophils (Lewis and Austen 1981, 1984; Peters et al. 1984, 1987). Detailed studies and reviews of the biochemistry of these mediators and their immunologically mediated reactions have been published (Lewis and Austen 1981, 1984; Lichtenstein et al. 1979; MacGlashan et al. 1982b; Paterson et al. 1976; Peters et al. 1984, 1987). Mast cells and basophils contain other important biochemicals. Both cells are characterized by their prominent complement of metachromatic cytoplasmic granules wherein reside various sulfated glycosaminoglycans (GAGS) (Galli et al. 1984). The most well-known of these is heparin (Metcalf et al. 1979). The various other GAGS and their distribution in basophils and mast cells of a variety of species form the subject of current intense investigation (reviewed in Galli et al. 1984). Early histochemical studies identified the presence of certain human mast cell enzymes. These included p-hexosaminidase, p-glucuronidase, and a trypsin-like esterase (Chiu and Lagunoff 1972; Glenner and Cohen 1960; Montagna 1957). More recently, biochemical analyses of isolated human lung mast cells have documented the presence of mast cell-associated tryptase, p-hexosam~nidase, p-glucuronidase, and aryl sulfatase (Schwartz et al. 1981 a, b, 1983). Human mast cells, as recognized in their fully differentiated forms, are wide spread in the connective tissues of virtually all organs in the body. They are concentrated in organs which are potential portals of entry for foreign agents, such as skin, lung, and gut. Mast cells also tend to be localized to perivascular structures in these organs. They can, however, be found throughout tissues, including within epithelia. Mast cells have the capacity to migrate and to proliferate and have been found in increased numbers in the tissues of patients with a wide variety of diseases. These disorders include chronic inflammatory disorders, cell-mediated immu nologic disorders, such as contact allergy, and neoplastic events of non-mast cell origin. Mast cell-specific disorders such as urticaria pigmentosa and systemic mastocytosis are characterized by extraordinary increases in mast cells in human tissues. For example, normal human skin mast cell counts vary between 5120 and 9472 per cubic millimeter (Mikhail and Miller-Milinska 1964). These values may be as high as 260000 to 380000 per cubic millimeter of skin in mastocytosis (Mikhail and Miller-Milinska 1964). Despite this, little to nothing is currently known regarding the functional role(s) of mast cells in these diseases. Moreover, little to nothing is known regarding the physiological role(s) of mast cells in health. Perhaps because so little is really known about the function of mast cells in health and disease, as well as the esthetic appeal of these cells, investigators continue to pursue their elusive functional capacities. It hardly seems likely that mast cells and basophils exist to plague people with sneezing, scratching, and wheezing episodes, or to kill us by anaphylaxis. Because knowledge of structure frequently precedes knowledge of function, and is essential for elucidating structure-function relationships, we herein review the structural information that is available for human mast cells. We first will itemize the differential diagnostic criteria for human basophils and human mast cells to assure the correct identification of these very similar cells. The in vivo ultrastruc tural anatomy of normal human mast cells from all available sites will be considered. Recent technological developments have made the isolation, purifi cation, and short-term culture of human mast cells from a variety of tissue sites possible (MacGlashan et al. 1982a). These sites include lung (Caulfield et al. 1980; Ishizaka et al. 1983; Paterson et al. 1976; Schulman et al. 1982, 1983), gut (Fox et al. 1985), and adenoids (Behrendt et al. 1978). Such preparations of human mast cells have made it possible to do structure-function studies on aliquots of the same mast cell preparations in vitro (Behrendt et al. 1978; Caulfield et al. 1980; Dvorak et al. 1983d, 1984, 1985d, 1986, 1987, 1988; Hammel et al. 1985). We will review the current state of these studies which includes analysis of mast cell organelles by a variety of ultrastructural techniques (Dvorak et al. 1983d, 1984; Hammel et al. 1985), of anaphylactic degranulation (Behrendt et al. 1978; Caulfield et al. 1980; Dvorak et al. 1984, 1985d), and recovery from degranulation (Dvorak et al. 1986, 1987, 1988). Other studies have utilized isolated skin slices for observations regarding mast cell release reactions (Greaves et al. 1972; Pearce et al. 1974; Tharp et al. 1983). Finally, we will review the available ultrastructural data in vivo implicating functioning mast cells in a variety of disorders (Galli et al. 1984). These studies include anaphylactic degranulation of mast cells in skin (Dvorak et al. 1976c; Kobayasi and Asboe-Hansen 1969a; Lepow et al. 1970; Yancey e,t al. 1985), heart (Dvorak 1986b), synovium (Wynne-Roberts and Anderson 1978), and piecemeal degranulation of mast cells (Galli and Dvorak 1984; Galli et al. 1984) in many human diseases involving a variety of organs (Colvin et al. 1974; Dvorak 1979; Dvorak and Dickersin 1979; Dvorak and Monahan 1985a; Dvorak et al. 1974, 1976c, 1978, 1980c,d, 1982; Galli and Dvorak 1984). 2 2 Ultrastructure of Human Mast Cells The ultrastructural anatomy of human mast cells has been described in numerous tissues. These sites include the following: skin (Asboe-Hansen 1971; Barnett 1975; Bowyer 1968; Burns and Hoak 1975; Dvorak et al. 1976c, 1980c, 1982; Fedorko and Hirsch 1965; Fujita et al. 1972; Hashimoto et al. 1966; Hibbs et al. 1960; Kobayasi and Asboe-Hansen 1969a, 1969b; Moriyasu and Yamura 1973; Naveh et al. 1975; Orr 1977; Pasyk et al. 1983; Trotter and Orr 1974; Yancey et al. 1985); lung (Brinkman 1968; Caulfield et al. 1980; Dvorak et al. 1983d, 1984, 1985d, 1986, 1987, 1988; Fox et al. 1981; Hammel et al. 1985; Kobayasi et al. 1968; Orr 1977; Parmley et al. 1975; Paterson et al. 1976; Ts'ao et al. 1977); bronchiole mucosa (Brinkman 1968; Trotter and Orr 1973; Ts'ao et al. 1977); nose (Taraska and Deno 1973; Trotter and Orr 1974); adenoids (Behrendt et al. 1978); lymph node (Parmley et al. 1975); bone marrow (Ts'ao et al. 1977); gingiva (Barnett 1973, 1975; Kobayasi et al. 1968; Weinstock and Albright 1967); stomach (Dobbins et al. 1969; Hibbs et al. 1960; Steer 1976); small bowel (Dobbins et al. 1969; Dvorak 1979; Dvorak and Dickersin 1979; Dvorak et al. 1978, 1980d; Rao 1973); colon (Dobbins et al. 1969; Fox et al. 1985); liver (Naveh et al. 1975); synovium (Wynne-Roberts and Anderson 1978); peripheral nerve (Pineda 1965); iris (Nii et al. 1974); heart (Dvorak 1986b); kidney (Colvin et al. 1974; Dvorak and Monahan 1985a). Human mast cells are readily recognized by electron microscopy. The only cell with which they may be confused is the basophil (see Sect. 3). Criteria for identification of mast cells in human tissues (Fig. 1) include a monolobed nucleus, surface architecture composed of narrow, elongated folds, the presence of typical cytoplasmic granules, and the absence of cytoplasmic glycogen aggregates. The cytoplasm also contains mito chondria, free ribosomes, intermediate filaments, and lipid bodies. Golgi areas are small in mature cells and membrane-bound ribosomes are rare in mature and imma ture cells. 1 I Human mast cells from various in vivo sources and in vitro experiments are presented following a variety of ultrastructural methods which illustrate different aspects of mast cell biology. The images, therefore, demonstrate variability, depending on the technique used. These methods and abbreviations for them are listed here. Abbreviations are used in individual legends to identify the specific method used to produce that image. All methods are described in primary publications in the references (Dvorak et al. 1972c, 1980b, 1981a, 1983b, 1983d, 1985b). 1. Osmium-collidine uranyl en bloc (OCUB) 2. Osmium potassium ferrocyanide (OPF) 3. Cationized ferritin (CF) 4. 3H-Arachidonic acid autoradiography eH-AA) 5. 3sS-Radiolabeled sulfur autoradiography eSS) 6. Endogenous peroxidase cytochemistry demonstrated with the substrate diaminobenzidine (DAB). 3 Fig. l. Human mast cell in vivo from a breast fibroadenoma shows a monolobed nucleus, narrow surface folds, and numerous cytoplasmic granules. OeUB, X 14500 Mature mast cell granules are smaller, vary more in shape, are more numerous, and generally contain more complex substructural patterns than the granules of basophils. Most observers have described several of the prevalent mast cell granule patterns in their individual works. Examination of all published micrographs as well as human mast cells from nearly all tissue sites (A.M. Dvorak, unpublished data) provides a working description of granule patterns. Different terms have been used by various authors to identify identical granule patterns. General agreement exists, however, among the published micrographs. We (Dvorak et al. 1984) and others refer to the four basic granule patterns of human mast cells as scrolls (Fig. 2A) (Asboe-Hansen 1971; Barnett 1973; Bowyer 1968; Brinkman 1968; Burns and Hoak 1975; Caulfield et al. 1980; Colvin et al. 1974; Dobbins et al. 1969; Dvorak 1979, 1986b; Dvorak and Dickersin 1979; Dvorak and Monahan 1985a; Dvorak et al. 1978, 1980c, d, 1982, 1983d, 1984, 1985d, 1986, 1988; Fox et al. 1981 ; Fox et al. 1985; Fujita et al. 1969; Hammel et al. 1985; Hashimoto et al. 1966; Kawanami et al. 1979; Kobayasi and Asboe-Hansen 1969b; Kobayasi et al. 1968; Moriyasu and Yamura 1973; Naveh et al. 1975; Nii et al. 1974; Orr 1977; Parmley et al. 1975; Pasyk et al. 1983; Paterson et al. 1976; Pineda 1965; Rao 1973; Steer 1976; Trotter and Orr 1974; Ts'ao et al. 1977; Weinstock and Albright 1967), cryst~ls (Asboe Hansen 1971; Burns and Hoak 1975; Caulfield et al. 1980; Dobbins et al. 1969; Dvorak and Dickersin 1979; Dvorak et al. 1976c, 1980c, d, 1982, 1983d, 1984, 1985d, 1986, 1988; Fedorko and Hirsch 1965; Fox et al. 1985; Hashimoto et al. 1966; Kawanami et al. 1979; Kobayasi and Asboe-Hansen 1969b; Kobayasi et al. 1968; Moriyasu and Yamura 1973; Orr 1973, 1977; Parmley et al. 1975; Pasyk et al. 1983; Trotter and Orr 1974; Ts'ao et al. 1977; Weinstock and Albright 1967), particles (Fig. 2B) (Asboe-Hansen 1971; Brinkman 1968; Colvin et al. 1974; Dobbins et al. 4