Summary
The ultrastructure of hepatic granulomas induced byCryptococcus (C.) neoformans was studied by a quick-freezing and deep-etching (QF-DE) method. Viable yeast cells were inoculated intravenously into rats and the livers were prepared for QF-DE replicas. Two cytoskeletal components were identified in the cytoplasm of macrophages composing the cryptococcal granulomas. These were: intermediate filaments, mainly located in the perinuclear region, and actin filaments, which were extensively decorated with myosin subfragment 1 (S1) and formed networks in the peripheral portion of the cytoplasm. In addition, two types of macrophage pseudopodia were observed in the granulomas. These were cobble stone-like pseudopodia at the yeast-macrophages contact areas, and thin, long and occasionally interdigitating pseudopodia in which actin filaments were consistently observed. Dense networks of actin filaments were also seen in pseudopodia protruding into the tight structure of the capsule ofC. neoformans. These results suggest a role for actin filaments as one of the main factors in the force generating system of the phagocytic process.
Similar content being viewed by others
References
Adams DO (1976) The granulomatous inflammatory response. A riview. Am J Pathol 84:164–192
Aggeler J, Werb Z (1982) Initial events during phagocytosis by macrophages viewed from outside and inside the cell: Membrane-particle interactions and clathrin. J Cell Biol 94:613–623
Baba T (1988) Electron microscopic cytochemical analysis of hepatic granuloma induced byCryptococcus neoformans. Mycopathologia 104:37–46
Baba T, Shiozawa N, Hotchi M, Ohno S (1991) Three-dimensional study of the cytoskeleton and multinucleate giant cells by quick-freezing and deep-etching method. Virchows Arch [B] 61:39–47
Baba T, Sakaguchi N, Hotchi M, Ohno S (1992) Three-dimensional study of epithelioid cells in muramyl dipeptide induced granulomas by quick-freezing and deep-etching method. Virchows Arch [B] 63:63–70
Bulmer GS, Tacker JR (1975) Phagocytosis ofCryptococcus neoformans by alveolar macrophage. Infect Immunol 11:73–79
Bulmer GS (1990) Twenty-five years withCryptococcus neoformans. Mycopathologia 109:111–122
Cain H, Kraus B (1981) Cytoskeleton in cells of the mononuclear phagocyte system. Virchows Arch [B] 36:159–176
Cain H, Krauspe R, Kraus B (1982) The cytoskeleton in activated and functionally disordered cells of the macrophage system. Pathol Res Pract 175:162–179
Diamond RD, May JE, Kane MA, Flank MM, Bennett JE (1974) The role of the classical and alternate complement pathways in host defenses againstCryptococcus neoformans. J Immunol 112:2260–2270
Emmons CW (1955) Saprophytic source ofCryptococcus neoformans associated with the pigeon (Columbia livia). Am J Hyg 62:227–232
Farmer SG, Komorowski RA (1973) Histologic response to capsule-deficientCryptococcus neoformans. Arch Pathol 96:383–387
Graeber MB, Streit WJ, Kreutzberg GW (1988) The microglial cytoskeleton: vimentin is localized within activated cells in situ. J Neurocytol 17:573–580
Hartwig JH, Davis WD, Stossel TP (1977) Evidence for contractile protein translocation in macrophage spreading, phagocytosis and phagolysosome formation. J Cell Biol 75:956–967
Hartwig JH, Yin HL (1988) The organization and regulation of the macrophage actin skeleton. Cell Motil Cytoskeleton 10:117–125
Hasenclever HF, Emmons CW (1963) The prevalence and mouse virulence ofCryptococcus neoformans strains isolated from urban areas. Am J Hyg 78:227–231
Heuser H, Kirschner M (1980) Filament organization revealed in platinum replicas of freeze-dried cytoskeleton. J Cell Biol 86:212–234
Ichikawa Y (1969) Differentiation of a cell line of myeloid leukemia. J Cell Physiol 74:223–234
Ishikawa H, Bischoff R, Holtzer H (1969) Formation of arrowhead complexes with heavy meromyosin in a variety of cell types. J Cell Biol 43:312–328
Kaplan MH, Rosen PP, Armstrong D (1977) Cryptococcosis in a cancer hospital: clinical and pathological correlates in fortysix patients. Cancer 39:2265–2274
Kalina M, Kletter Y, Aronson M (1974) The interaction of phagocytes and large-sized parasiteCryptococcus neoformans: cytochemical and ultrastructural study. Cell Tissue Res 152:165–174
Kovacs JA, Kovacs AA, Polis M, Wright WC, Gill VJ, Tuazon CU, Gelman EP, Lane HC, Longfield R, Overturf G, Macher AM, Fauci AS, Parrillo JE, Bennett JE, Masur H (1985) Cryptococcosis in the acquired immunodeficiency syndrome. Ann Intern Med 103:533–538
Kozel TR (1977) Non-encapsulated variant ofCryptococcus neoformans II surface receptors for cryptococcal polysaccharide and their role in inhibition of phagocytosis by polysaccharide. Infect Immunol 16:99–106
Kozel TR, Gotschlich EC (1982) The capsule ofCryptococcus neoformans passively inhibits phagocytosis of the yeast by macrophages. J Immunol 129:1675–1680
Kwon-chung KJ (1977) Cryptococcosis. In: Emmons CW, Binford CH, Utz JP, Kwon-chung KJ (eds) Medical mycology, 3rd edn. Lea and Febiger, Philadelphia, pp 206–229
Lazarides E (1980) Intermediate filaments as mechanical integrators of cellular space. Nature 283:249–256
Lehto V-P, Virtanen I, Kurki P (1978) Intermediate filaments anchor the nuclei in nuclear monolayers of human fibroblast. Nature 272:175–177
Marchevsky A, Rosen MJ, Chrystal G, Kleinerman J (1985) Pulmonary complications of the acquired immunodeficiency syndrome: a clinicopathologic study of 70 cases. Hum Pathol 16:659–670
Marugg RA, Gehr P, de Leeuw M (1990) Secondary lysosomes as an integral part of the cytoskeleton: a morphological study in rat Kupffer cells. J Struct Biol 105:146–153
Miyaji M, Nishimura K (1985) Defensive role in granuloma against fungal infections. In: Arai T (eds) Filamentous microorganism. Japan Scientific Societies Press, Japan, pp 263–277
Naramoto A, Ohno S, Furuta K, Itoh N, Nakazawa K, Nakano M, Shigematsu H (1991) Ultrastructural studies of hepatocyte cytoskeletons of phalloidin-treated rats by quick-freezing and deep-etching method. Hepatology 13:222–229
Ohno S (1985) Immunocytochemical study on the cytoplasmic side of cell membranes infected with vesicular stomatitis virus by quick-freezing and deep-etching replica method. Histochemistry 82:565–575
Ohno S, Fujii Y (1990) Three-dimensional and histochemical studies of peroxisomes in cultured hepatocytes by quick-freezing and deep-etching method. Histochem J 22:143–154
Osumi M, Baba M, Naito N, Taki A, Yamada N, Nagatani T (1988) High resolution, low voltage scanning electron microscopy of uncoated yeast cells fixed by the freeze-substitution method. J Electron Microsc 37:17–30
Reaven EP, Axline SG (1972) Subplasmalemmal microfilaments and microtubules in resting and phagocytizing cultivated macrophages. J Cell Biol 59:12–27
Rinnerthaler G, Herzog M, Klappacher M, Kunda H, Small JV (1991) Leading edge movement and ultrastructure in mouse macrophages. J Struct Biol 106:1–16
Rippon JW (1982) Medical mycology: The pathogenic fungi and the pathogenic actinomycetes, 2nd edn. Saunders Company, Philadelphia London Tront, pp 532–558
Sakaguchi N, Baba T, Ohno S (1993) Ultrastructural study ofCryptococcus neoformans by quick-freezing and deep-etching method. Mycopathologia (in press)
Senda T, Fujita H, Ban T, Zhong C, Ishimura K, Kanda K, Sobue K (1989) Ultrastructural and immunocytochemical studies on the cytoskeleton in the anterior pituitary of rats, with special regard to relationship between actin filaments and secretory granules. Cell Tissue Res 258:25–30
Toyohara A, Inaba K (1989) Transport of phagosomes in mouse peritoneal macrophages. J Cell Sci 94:143–153
Trotter JA (1981) The organization of actin in spreading macrophages. Exp Cell Res 132:235–248
Tsuru A, Nakamura N, Takayama E, Suzuki Y, Hirayoshi K, Nagata K (1990) Regulation of the expression of vimentin gene during the differentiation of mouse myeloid leukemia cells. J Cell Biol 110:1655–1664
Yin HL, Hartwig JH (1988) The structure of the macrophage actin skeleton. J Cell Sci Suppl 9:169–184
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Sakaguchi, N. Ultrastructural study of hepatic granulomas induced byCryptococcus neoformans by quick-freezing and deep-etching method. Virchows Archiv B Cell Pathol 64, 57–66 (1993). https://doi.org/10.1007/BF02915096
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02915096