Abstract
Crude extracts of the lipophilic yeast Malassezia furfur were obtained from 2, 6, 10 and 28 day old cultures. The in vitro cultivation periods corresponded, respectively, to the lag phase, middle of the log phase, end of log phase and the decline phase of the growth curve, which was based on viable cell counts obtained with a fluorescent viability test. Biochemical analyses showed that the protein and carbohydrate contents were greater in day 10 extracts. Seventy patients with different allergic manifestations and 30 healthy volunteers were skin prick tested using the extracts. Of these, thirteen (18.57%) patients gave positive responses. SDS PAGE gradient electrophoretic profiles of the preparations indicated that the 28 day extracts contained the greatest number of protein bands with molecular weights ranging mostly between 30 and 94 kDa. Immunoblots incubated with individual patient sera showed that four IgE binding M. furfur allergens of approximately 88, 61, 52 and 39 kDa were present in the 28 day extracts. The components identified could be used for detecting IgE mediated responses to M. furfur among individuals affected with different allergic conditions.
Similar content being viewed by others
References
Gordon MA. Liphophilic yeast-like organisms associated with tinea versicolor. J Invest Dermatol 1951; 17: 267–272.
Faergemann J, Arly R, Maibach HI. Growth and filament production of Pityrosporum orbiculare and P. ovale on human stratum corneum in vitro. Acta Dermato-Venereologica 1983; 63: 388–392.
Doekes G. Biochemical characteristics of Pityrosporum allergens. Allergy 1994; 49: 198–199.
Yunginger JW. Allergens: recent advances. Pediatric Clin N Am 1988; 35: 981–993.
Auklust L, Borch SM, Aas K, Stering I. Allergens in Cladosporium herbarum: advances in characterization of Cladosporium herbarum allergens. Int Arch Allergy Appl Immunol 1993; 69: 122–127.
Harvey C, Longbottom JL. Characterization of a major antigenic component of Aspergillus fumigatus. Clin Exp Immunol 1986; 65: 206–214.
Alonso A, Scavini LM, Mouchian K, Rodriguez SM, Iraneta SG. Antigenicity of Penicillium notatum in animals and in atopic patients. Allergol Immunopathol 1990; 816: 301–307.
Wessels M, Doekes G, Van Ieperen-Van Kijk A, Koers W, Young E. IgG antibodies to Pityrosporum ovale in atopic dermatitis. Br J Dermatol 1991; 125: 227–232.
Cunningham A, Inghan E, Gowland G. Humoral responses to Malassezia furfur serovars A, B and C in normal individuals of various ages. Br J Dermatol 1992; 127: 476–481.
Nordvall Sl, Lindgren L, Johansson SGO, Johansson S, Petrini B. IgE antibodies to Pityrosporum orbiculare and Staphylococcus aureus in patients with very high serum total IgE. Clinical Exp Allergy 1992; 22: 756–761.
Broberg A, Faergemann J, Johansson S, Johansson SGO, Strannegard IL, Svejgaard, E. Pityrosporum ovale and atopic dermatitis in children and young adults. Acta Derm Venereol 1992; 72: 187–192.
Zargari A, Harfast B, Johansson S, Johansson SGO, Scheynius A. Identification of allergen components of the opportunistic yeast Pityrosporum orbiculare by monoclonal antibodies. Allergy 1994; 49: 50–56.
Yasueda H, Okado TH, Saito A, Uchida K, Kuroda M, OnishI Y, Takahashi K, Yamaguchi H, Takesako K, Akiyama K. Identification and cloning of two novel allergens from the lipophilic yeast, Malassezia furfur. Biochemical and Biophisical Research Communications 1998; 248: 240-244.
Schimdt M, Zargari A, Holt P, Lindbon L, Hellman ULF, Whitley P, Van Der Ploeg I, Harfast B, Scheynius A. The complete cDNA sequence and expression of the first major allergenic protein of Malassezia furfur, Mal f 1. Eur J Biochem 1999; 246: 180–185.
Onishi Y, Kuroda M, Yasueda H, Saito A, Koyama ES, Tunasawa S, Okado TH, Yagihara T, Uchida K, Yamaguchi H, Akiyama K, Kato I, Takesako K. Two-dimensional eletrophoresis of Malassezia allergens for atopic dermatitis and isolation of Mal f 4 homologs with mitochondrial malate deydrogenase. Eur J Biochem 1999, 261: 148–154.
Lindbong M, Magnusson CG, Zargari A, Schimdt M, Scheynius A, Crameri R, Whitley P. Selective cloning of allergens from the skin colonizing yeast Malassezia furfur by phage surface display technology. J Invest Dermatol 1999, 113: 156–161.
Guillot J, Guého E, Lesourd M, Midgley G, Ch,vrier G, Dupond B. Identification of Malassezia species. A practical approach. J Mycol Med 1996; 6: 103–110.
Mayser P, Haze P, Papavassilis C, Pickel M, Gruender K, GuÉho E. Differentiation of Malassezia species: Selectivity of Cremophor El, castor oil and ricinoleic acid for M. furfur 1997; 137: 208–213.
Calich VLG, Purchio A, Paula CR. A new fluorescent viability test for fungi cells. Mycopathologia 1978; 66: 175–177.
Dacie JV, Lewis SM. Pratical haematology. 3a ed., London, J&A Churchill, 1963; 18 p.
Mendes E. Alergia no Brasil. Sao Paulo, Manole, 1989: 221 p.
Lowry OH, Rosebrough NJ, Farr AL, Randall, RJ. 1951. Protein measurement with The Folin-phenol reagent. J Biol Chem 1951; 193: 265.
Scott TA, Melvin EH. Determination of dextran with anthrone. Anal Chem 1953, 25: 1656–1661.
Nichols AV, Klaus RM, Nushiner TA. Nondenaturing polyacrilamide gradiente gel electrophoresis. Methods Enzymol 1986; 128: 417-431.
Camargo ZP, Unterkircher C, Campoy SP, Travassos LR. Production of Paracoccidioides brasiliensis exoantigens for immunodiffusion tests. J Clin Microbiol 1988; 26: 2147–2151.
Ran Y, Yoshiike T, Ogawa H. Lipase of Malassezia furfur: some properties and their relationship to cell growth. J Med and Veter Mycology 1993; 31: 77–85.
Morris EO. Yeast growth. In: Cook AH ed. The Chemistry and Biology of Yeast. New York 1958: 251–322.
San-Blas F, Cova LJ. Growth curves of the yeast like form of Paracoccidioides brasiliensis. Sabouraudia 1975; 13: 22–29.
Morris EO. Composition and structure of yeast cell walls. In: Rose A, Harrison JS, eds. The Yeast. 2A. ed. London, Academic Press, V. 3, 1985; 317–366.
Rokugo M, Tagami H, Usuda Y, Tomita Y. Contact sensitivity to Pityrosporum ovale in patients with atopic dermatitis. Arch of Dermat 1990; 126: 627–632.
Kieffer M, Bergbrant I.M, Faergemann J. Immune reactions to Pityrosporum ovale in patients with atopic and seborreheic dermatitis. J Am Acad Dermatol 1990; 22: 739–742.
Broberg A. Pityrosporum ovale in healthy children, infantile seborrhoeic dermatitis and atopic dermatitis. Acta Derm Venereol Suppl 1995; 191: 1–47.
Huang X, Johansson SGO, Zargari A, Nordvall SL. Allergen cross-reactivity between Pityrosporum orbiculare and Candida albicans. Allergy 1995, 50: 648–656.
Carrera MD, Soto C, Gusmán G. Cuantificatión de Extractos Alêrgenicos en Unidades deMasa. Ed. Alergia e Immunologia Abelló, S.A., Madrid 1993.
Portnoy J, Olson I, Pacheco F, Banes C. Affinity purification of major Alternaria allergen using a monoclonal antibody. Ann Allergy 1990; 65: 109–114.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Gandra, R., Melo, T., Matsumoto, F. et al. Allergenic evaluation of Malassezia furfur crude extracts. Mycopathologia 155, 183–189 (2003). https://doi.org/10.1023/A:1021181711225
Issue Date:
DOI: https://doi.org/10.1023/A:1021181711225