, Volume 165, Issue 2, pp 73–80 | Cite as

Immunohistochemical and immunocytochemical detection of SchS34 antigen in Stachybotrys chartarum spores and spore impacted mouse lungs

  • Thomas G. Rand
  • J. David Miller


The purpose of this study was to evaluate the distribution of a 34 kD antigen isolated from S. chartarum sensu lato in spores and in the mouse lung 48 h after intra-tracheal instillation of spores by immuno-histochemistry. This antigen was localized in spore walls, primarily in the outer and inner wall layers and on the external wall surfaces with modest labelling observed in cytoplasm. Immuno-histochemistry revealed that in spore impacted mouse lung, antigen was again observed in spore walls, along the outside surface of the outer wall and in the intercellular space surrounding spores. In lung granulomas the labelled antigen formed a diffusate, some 2–3× the size of the long axis of spores, with highest concentrations nearest to spores. Collectively, these observations indicated that this protein not only displayed a high degree of specificity with respect to its location in spores and wall fragments, but also that it slowly diffuses into surrounding lungs.


Stachybotrys chartarum Antigen Spores Mouse lung Immunohistochemistry Antigen processing 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.



We thank C. Leggiadro NRC Institute of Marine Biosciences for assistance and excellent technical support. Jiangping Xu prepared the antibodies. This study was supported by a Natural Sciences and Research Council discovery grant administered to TGR and an NSERC IRC to JDM.


  1. 1.
    Health Canada. Fungal contamination in public buildings: health effects and investigation methods. Health Canada, Ottawa, ON. 2004. ISBN 0-662-37432-0.Google Scholar
  2. 2.
    NAS. Damp indoor air spaces and health. Washington, DC: Institute of Medicine, National Academies Press; 2004.Google Scholar
  3. 3.
    WHO. Health and environmental briefing #42 for local health officials. World Health Organization European Centre for Environment and Health, Hermann-Ehlers-Strase, Bonn, Germany, 2004.Google Scholar
  4. 4.
    Jaakkola JJ, Hwang BF, Jaakkola N. Home dampness and molds, parental atopy, and asthma in childhood: a six-year population-based cohort study. Environ Health Perspect. 2005;113:357–61.PubMedGoogle Scholar
  5. 5.
    Douwes J, Thorne P, Pearce N, Heederick N. Bioaerosol health effects and exposure assessment: progress and prospects. Ann Occup Hyg 2003;47:187–200.PubMedCrossRefGoogle Scholar
  6. 6.
    Schram-Bijkerk D; Doekes G, Douwes J, Boeve M, Riedler J, Ueblagger E, von Mutius E, Benz MR, Pershagen G, van Hage M, Scheynius A, Braun-Fahrlander C, Waser M, Brunekreef B. Bacterial and fungal agents in house dust and wheeze in children: the PARSIFAL study. Clin Exp Allergy 2005;35:1272–8.CrossRefGoogle Scholar
  7. 7.
    NAS. Clearing the air. Washington, DC: National Academy of Science, National Academy Press; 2000.Google Scholar
  8. 8.
    Van Emon JM, Reed AW, Yike I, Vesper SJ. Measurement of Stachylysin™ in serum to quantify human exposures to the indoor mold Stachybotrys chartarum. J Occup Environ Med. 2003;45:582–91.PubMedCrossRefGoogle Scholar
  9. 9.
    Vesper SJ, Magnuson S, Dearborn DG, Yike I, Haugland RA. Initial characterization of the hemolysin stachylysin from Stachybotrys chartarum. Infect Immun. 2001;69: 912–6.PubMedCrossRefGoogle Scholar
  10. 10.
    Yike I, Distler AM, Ziady AG, Dearborn DG. Mycotoxin adducts on human serum albumin: biomarkers of exposure to Stachybotrys chartarum. Environ Health Perspect 2006;114:1221–6.PubMedCrossRefGoogle Scholar
  11. 11.
    Schemechel D, Simpson JP, Beezhold D, Lewis DM. The development of a species specific immunodiagnsostics for Stachybotrys chartarum: the role of cross reactivity. J Immunol Methods 2006;309:150–9.CrossRefGoogle Scholar
  12. 12.
    Xu J, Jensen JT, Liang Y, Miller JD. The biology and immogenicity of the 34 kDa antigen of Stachybotrys chartarum sensu lato. Int Biodeterior Biodegrad 2007 (in press).Google Scholar
  13. 13.
    Xu J, Liang Y, Miller JD. Characterization of monoclonal antibodies of SchS protein from Stachybotrys chartarum and their application in measurement of S. chartarum antigen by capture ELISA (in press).Google Scholar
  14. 14.
    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. Chem Phys Lipids 2001;110:1–10.CrossRefGoogle Scholar
  15. 15.
    Rand TG, Mahoney M, White K, Oulton M. Microanatomical changes in alveolar type II cells in juvenile mice intratracheally exposed to Stachybotrys chartarum spores and toxin. Toxicol Sci 2002;65:239–45.PubMedCrossRefGoogle Scholar
  16. 16.
    Gregory L, Pestka JJ, Dearborn DG, Rand TG. Localization of satratoxin-G in Stachybotrys chartarum spores and spore-impacted mouse lung using immunocytochemistry. Toxicol Pathol 2004;32:26–34.PubMedCrossRefGoogle Scholar
  17. 17.
    Hudson B, Flemming J, Sun G, Rand TG. Comparison of immunomodulator mRNA expression and concentration in lungs of Stachybotrys chartarum spore exposed mice. J Toxicol Environ Health A 2005;68:1321–35.PubMedCrossRefGoogle Scholar
  18. 18.
    Flemming J, Hudson B, Rand TG. Comparison of inflammatory and cytotoxic lung responses in mice after intratracheal exposure to spores of two different Stachybotrys chartarum strains. Toxicol Sci 2004;78: 267–76.PubMedCrossRefGoogle Scholar
  19. 19.
    Rand TG, Flemming J, Miller JD, Womiloju TO. Comparison of inflammatory responses in mouse lungs exposed to atranones A and C from Stachybotrys chartarum. J Toxicol Environ Health A 2006;69:1239–51.PubMedCrossRefGoogle Scholar
  20. 20.
    Rand TG, Giles S, Flemming J, Miller JD, Puniani E. Inflammatory and cytotoxic responses in mouse lungs exposed to purified toxins from building isolated Penicillium brevicompactum Dierckx and P. chrysogenum Thom. Toxicol Sci 2005;87:213–22.PubMedCrossRefGoogle Scholar
  21. 21.
    Okudaira M, Kurata H, Sakabe F. Studies on the fungal flora in the lung of human cases. A critical survey in correction of the pathogenesis of opportunistic fungus infections. Mycopathologia 1977;61:3–18.PubMedCrossRefGoogle Scholar
  22. 22.
    Hung LL, Miller JD, Dillon HK, editors. Field guide for the determination of biological contaminants in environmental samples, 2nd ed. American Industrial Hygiene Association, Fairfax, VA; 2005. p. 93–128.Google Scholar
  23. 23.
    Nolard N. Moulds and respiratory allergies. Expressions 1997;5:7–9.Google Scholar
  24. 24.
    Jakab GJ, Hmieleski RR, Zarba A, Hemenway DR, Groopman JD. Respiratory aflatoxicosis: suppression of pulmonary and systemic host defences in rats and mice.Toxicol. Appl Pharmacol 1994;125:198–205.CrossRefGoogle Scholar
  25. 25.
    Richard JL, Thurston JR. Effect of aflatoxin on phagocytosis of Aspergillus fumigatus spores by rabbit alveolar macrophages. Appl Micro.1975;30:44–7.Google Scholar
  26. 26.
    Sorenson WG. Fungal spores: hazardous to health? Environ Health Perspect 1999; 107 s.3:469–72.Google Scholar
  27. 27.
    Olenchock SA, Green FH, Mentnech MS, Mull JC, Sorenson WG. In vivo pulmonary response to Aspergillus terreus spores. Comp Immunol Microbiol Infect Dis 1983;6:67–80.PubMedCrossRefGoogle Scholar
  28. 28.
    McDowell EM, Trump BF. Histological fixatives suitable for diagnostic light and electron microscopy. Arch Pathol Lab Med 1976;100:405–14.PubMedGoogle Scholar
  29. 29.
    Bozzola JJ, Russell LD. Electron microscopy. Boston, MA: Jones and Bartlett Publishers; 1992.Google Scholar
  30. 30.
    CCAC. Guide to the care and use of experimental animals, vol 1. Canadian Council on Animal Care, Bradda Printing Services Inc., Ottawa, ON; 1993.Google Scholar
  31. 31.
    Foto M, Vrijmoed LLP, Miller JD, Ruest K, Lawton M, Dales RE. Comparison of airborne ergosterol, glucan and Air-O-Cell data in relation to physical assessments of mold damage and some other parameters. Indoor Air 2005;15:257–66.PubMedCrossRefGoogle Scholar
  32. 32.
    Toivola M, Nevalainen A, Alm S. Personal exposures to particles and microbes in relation to microenvironmental concentrations. Indoor Air 2004;14:351–9.PubMedCrossRefGoogle Scholar
  33. 33.
    Gorny RL, Reponen T, Willeke K, Schmechel D, Robine E, Boissier E, Grinshpun SA. Fungal fragments as indoor air biocontaminants. Appl Environ Microbiol 2002;68: 3522–31.PubMedCrossRefGoogle Scholar
  34. 34.
    Gregory L, Rand TG, Dearborn D Yike I, Vesper S. Immunocytochemical localization of a hemolysin-like protein in Stachybotrys chartarum spores and spore-impacted mouse and rat lung tissues. Mycopathologia 2002;56:77–85.Google Scholar
  35. 35.
    Yeates DB, Daza AV, Mussatto DJ. Bronchial and alveolar allergen-induced anaphylaxis and the stimulation of bronchial mucociliary clearance in ragweed-sensitized dogs. Proc Assoc Am Physicians 1997;109:440–52.PubMedGoogle Scholar
  36. 36.
    Sturm R, Hofman W. A multicompartment model for slow bronchial clearance of insoluble particles- extension of the ICRP human respiratory tract models. Radiot Prot Dosimetry 2006;118:384–94.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  1. 1.Department of BiologySaint Mary’s UniversityHalifaxCanada
  2. 2.Ottawa-Carleton Institute of ChemistryCarleton UniversityOttawaCanada

Personalised recommendations