Skip to main content
SpringerLink
Log in

Immunocytochemical localization of stachylysin in Stachybotrys chartarum

  • Published:
Mycopathologia Aims and scope Submit manuscript

Abstract

Stachylysin is a proteinaceous hemolytic agent that is produced by Stachybotrys chartarum. Stachylysin was found, using immunohistochemical and immunocytochemical methods, to be localized in S. chartarum spores/mycelia primarily in the inner wall suggesting that it is constitutively produced. Spores instilled in mouse or rat lung tissues resulted in granuloma formation, which showed the highest stachylysin concentration in the inner wall of the spore and near the spore, with less at distance indicating that it had diffused out from the spore. The in vitro high stachylysin producing strain (58-06) was also highest in vivo, based on immunohistochemistical staining. More stachylysin was observed in the mouse lung tissue at 72 h than at 24 h indicating that production/release is a relatively slow process. The localization of stachylysin in macrophage phagolysosomes suggests that these cells may be involved with hemolysin inactivation. This would be consistent with what is known about asp-hemolysin produced by Aspergillus fumigatus.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Bhakdi S, Bayley H, Valeva I, Walev A,Walker B, Weller U, Kehoe M, Palmer M. Staphylococcal alpha-toxin, streptolysin-O, and Escherichia coli hemolysin prototypes of pore-forming bacterial cytolysins. Arch. Microbiol. 1996; 165: 73–79.

    Google Scholar 

  • Bhakdi S, Tranum-Jensen J. Alpha-toxin of Staphylococcus aureus. Microbiol. Rev. 1991; 55: 733–751.

    Google Scholar 

  • Bozzola JJ, Russell LD. Electron Microscopy. Boston: Jones and Bartlett Publishers, 1992.

    Google Scholar 

  • Cavalieri SJ, Bohach GA, Snyder IS. Escherichia coli a-hemolysin characteristics and probable role in pathogenicity. Microbiol Rev 1984; 48: 326–343.

    Google Scholar 

  • CCAC. Guide to the care and use of experimental animals. Volume 1. Ottawa: Bradda Printing Services Inc., Ottawa, 1993.

    Google Scholar 

  • Davies P. Quantitative morphology of the lung. A practical Manual. Harvard Medical School Publication, 1980.

  • Dearborn DG, Yike I, Sorenson WG, Miller MJ, Etzel RA. Overview of investigations into pulmonary hemorrhage among infants in Cleveland, Ohio. Environ Health Persp 1999; 107: 495–499.

    Google Scholar 

  • Deacon JW. Introduction to Modern Mycology. Boston: Blackwell Scientific Publications, 1984.

    Google Scholar 

  • Ebina K, Yokota K, Sakaguchi O. Studies on the toxin of Aspergillus fumigatus. XIV. Relationship between Asp-hemolysin and experimental infection of mice. Jpn J Med Mycol 1982; 23: 246–252.

    Google Scholar 

  • Ebina K, Yokota K, Sakaguchi O. Studies on the toxin of Aspergillus fumigatus. XVI. Biological properties of Asp-hemolysin as a parasitic factor. Jpn J Med Mycol 1983; 24: 245–252.

    Google Scholar 

  • Ebina K, Sakagami H, Yokota K, Kondo H. Cloning and nucleotide sequence of cDNA encoding Asp-hemolysin from Aspergillus fumigatus. Biochimia et Biophysica Acta. 1994; 1219: 148–150.

    Google Scholar 

  • Elidemir O, Colasurdo GN, Rossmann SN, Fan LL. Isolation of Stachybotrys from the lung of a child with Pulmonary Hemosiderosis. Pediatrics 1999; 102: 964–966.

    Google Scholar 

  • Etzel RA, Montana E, Sorenson WG, Kullman GJ, Allan TM, Dearborn, DG. Acute pulmonary hemorrhage in infants associated with exposure to Stachybotrys atra and other fungi. Arch Pediatr Adolesc Med 1998; 152: 757–762.

    Google Scholar 

  • Flappan SM, Portnoy J, Jones P, Barnes C. Infant pulmonary hemorrhage in a suburban home with water damage and mold (Stachybotrys atra). Envir Health Persp 1999; 107: 927-930.

    Google Scholar 

  • Forgacs J. Stachybotryotoxicosis. In Kadis S, Ciegler A, Ajl SJ, eds. Microbial Toxins, Vol. VIII. New York, NY: Academic Press, Inc., 1972; 95–128.

    Google Scholar 

  • Gibson RL, Nizet V, Rubens CE. Group B streptococcal ?-hemolysin promotes injury of lung microvascular endothelial cells. Ped Res 1999; 45: 626–634.

    Google Scholar 

  • Happerfield LC. Assessment of oestrogen and progesterone receptor antibodies in formalin-fixed routinely processed paraffin-wax embedded tissue. J Cell Path 1996; 1: 170–178.

    Google Scholar 

  • Hintikka E.-L. Human stachybotrytoxicosis. In Willie TD, Morehouse LG, eds. Mycotoxic Fungi, Mycotoxins, Mycotoxicoses, An Encyclopedic Handbook. Vol 3, New York: Marcel Dekker, Inc. 1978; 87–89.

    Google Scholar 

  • Humason GL. Animal Tissue Techniques. San Francisco: W.H. Freeman and Co., 1976.

    Google Scholar 

  • Jarvis BB, Sorenson WG, Hintikka E.-L, Nikulin M, Zhou Z, Jiang J, Wang S, Hinkley S, Etzel RA, Dearborn DG. Study of toxin production by isolates of Stachybotrys atra and Memnoniella echinata isolated during a study of pulmonary hemosiderosis in infants. Appl Environ Microbiol 1998; 64: 3620–3625.

    Google Scholar 

  • Johnson P, Lindberg M, Haraldsson I, Waldstrom T. Virulence of Staphylococcus aureus in a mouse mastitis model: studies of alpha hemolysin, coagulase and protein A as possible virulence determinants with protoplast fusion and gene cloning. Infect Immun 1985; 49: 765–769.

    Google Scholar 

  • MaCrae KC, Rand TG, Shaw RA, Mason C, Oulton MR, Hastings C, Cherlet T, Thliveris JA, Mantsch HH, MacDonald J, Scott JE. Analysis of pulmonary surfactant by Fourier-transform infrared spectroscopy following exposure to Stachybotrys chartarum (atra) spores. Chemistry and Physics of Lipids 2001; 110: 1–10.

    Google Scholar 

  • Manns JM, Mosser DM, Buckley HR. Production of a hemolytic factor by Candida albicans. Infect Immun 1994; 62: 5154–5156.

    Google Scholar 

  • Mason CD, Rand TG, Oulton M, MacDonald JM, Scott JE. Effects of Stachybotrys chartarum (atra) Conidia and Isolated Toxin on Lung Surfactant Production and Homeostasis. Natural Toxins 1998; 6: 27–33.

    Google Scholar 

  • Mason CD, Rand TG, Oulton M, MacDonald J. The effect of Stachybotrys chartarum spores and an isolated trichothecene, isosatratoxin F, on convertase activity in mice. Toxicology and Applied Pharmacology 2001; 172: 21–28.

    Google Scholar 

  • McDowell EM, Trump BF. Histological fixatives suitable for diagnostic light and electron microscopy. Arch Pathol Lab Med 1976; 100: 405–414.

    Google Scholar 

  • Nikulin M, Reijula K, Jarvis BB, Hintikka E.-L. Experimental lung mycotoxicosis in mice induced by Stachybotrys atra. Int J Exp Path 1996; 77: 213–218.

    Google Scholar 

  • Nikulin M, Reijula K, Jarvis B, Veijalainen P, Hintikka E-L. Effects of intranasal exposure to Stachybotrys atra. Fundam Appl Toxicol 1997; 35: 182–188.

    Google Scholar 

  • Nizet V, Gibson RL, Chi EY, Framson PE, Hulse M, Rubens CE. Group B streptococcal beta-hemolysin expression is associated with injury of lung epithelial cells. Infect Immun 1996; 64: 3818–3826.

    Google Scholar 

  • Nomura T, Fujii Y, Okamota K. Secretion of hemolysin of Aeromonas sobria as protoxin and contribution of the propeptide region removed from the protoxin to the proteolytic stability of the toxin. Microbiol Immunol 1999; 43: 29–38.

    Google Scholar 

  • Ou Said A M, Contrepois MG, Vartanian MD, Girardeau J. Virulence factors and markers in Escherichia coli from calves with bacteremia. Am J Vet Res 1988; 49: 1657–1660.

    Google Scholar 

  • Rand TG, Mahoney M, White K, Oulton M. Microanatomical changes associated with alveolar type II cells in juvenile mice exposed to Stachybotrys chartarum and isolated toxin. Toxicological Sciences. 2002; in press.

  • Rao CY, Burge HA, Brain JD. The time course of responses to intratracheally instilled toxic Stachybotrys chartarum spores in rats. Mycopathologia 2000; 149: 27–34.

    Google Scholar 

  • Sakaguchi O, Yokota K. Studies on the toxin of Aspergillus fumigatus III. Fraction causing dermonecrosis and hemorrhage-like changes. Japan J Bacteriol 1972; 27: 649–655.

    Google Scholar 

  • Sakaguchi O, Shimada H, Yokota K. Purification and characteristics of hemolytic toxin from Aspergillus fumigatus. Japan J Med Sci Biol 1975; 28: 328–331.

    Google Scholar 

  • Seeger W, Obernitz R, Thomas M, Walmrath D, Suttorn N, Holland IB, Grimminger F, Eberspacher B, Hugo F, Bhakdi S. Lung vascular injury after administration of viable hemolysin-forming Escherichia coli in isolated rabbit lungs. Am Rev Resp Dis 1991; 143: 797–785.

    Google Scholar 

  • Sorenson WG, Lewis DM. Organic dust toxic syndrome. In Esser K, Lempke PA, eds. The Mycota, Vol. VII. Animal and Human Relationships. Springer Verlag, Berlin. 1996; 159–172.

    Google Scholar 

  • Sumarah MW, Rand TG, Mason CD, Oulton M, MacDonald J, Anthes M. Effects of Stachybotrys chartarum spores and toxin on mouse lung surfactant phospholipid vomposition. In: Johanning E, ed. Bioaerosols, Fungi and Mycotoxins: Health Effects, Assessment, Prevention and Control. Eastern New York Occupational and Environmental Health Center, Albany, New York, 1999; 444–452.

    Google Scholar 

  • Vesper SJ, Dearborn DG, Yike I, Sorenson WG, Haugland RA. Hemolysis, toxicity and randomly amplified polymorphic DNA analysis of Stachybotrys chartarum strains. Applied and Environ. Microbiology 1999; 65(7): 3175–3181.

    Google Scholar 

  • Vesper SJ, Dearborn DG, Elidemir O, Haugland RA. Quantification of siderophore and hemolysin from Stachybotrys chartarum strains including a strain isolated from the lung of a child with pulmonary hemorrhage and haemosiderosis. Applied and Environ Microbiology. 2000; 66(6): 2678–2681.

    Google Scholar 

  • Vesper SJ, Magnuson DG, Dearborn G, Yike I, Haugland RA. Initial characterization of the hemolysin stachylysin from Stachybotrys chartarum. Infect. Immun. 2001; 69: 912–916.

    Google Scholar 

  • Yike I, Allan T, Sorenson WG, Dearborn DG. Highly sensitive protein translation assay for trichothecene toxicity in airborne particulates. Comparison with cytotoxicity assays. Appl Environ Microbiol 1999; 65: 88–94.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biology, Saint Mary's University, Halifax, Nova Scotia, B3H 3C3

    L. Gregory & T.G. Rand

  2. Department of Pediatrics, Case Western University, Cleveland, OH

    D. Dearborn & I. Yike

  3. National Exposure Research Laboratory, US Environmental Protection Agency, Cincinnati, OH

    S. Vesper

Authors
  1. L. Gregory
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. T.G. Rand
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. D. Dearborn
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. I. Yike
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. S. Vesper
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gregory, L., Rand, T., Dearborn, D. et al. Immunocytochemical localization of stachylysin in Stachybotrys chartarum . Mycopathologia 156, 109–117 (2003). https://doi.org/10.1023/A:1022968121285

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1022968121285

Search

Navigation