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Microbiology 151 (2005), 291-299; DOI  10.1099/mic.0.27433-0
© 2005 Society for General Microbiology

Melanization of Penicillium marneffei in vitro and in vivo

Sirida Youngchim1,2, Roderick J. Hay3 and Andrew J. Hamilton1

1 Dermatology Department, St Johns Institute of Dermatology, 5th Floor Thomas Guy's House, Guy's Hospital, Kings and St Thomas' Medical Schools, London SE1 9RT, UK
2 Microbiology Department, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand
3 Faculty of Medicine and Health Sciences, Queen's University, Belfast, Northern Ireland, UK

Correspondence
Sirida Youngchim
syoungch{at}mail.med.cmu.ac.th

Melanins are found universally in nature and are implicated in the pathogenesis of several important human fungal pathogens. This study investigated whether the conidia and the yeast cells of the thermally dimorphic fungal pathogen Penicillium marneffei produce melanin or melanin-like compounds in vitro and during infection. Treatment of conidia with proteolytic enzymes, denaturant and concentrated hot acid yielded dark particles that were similar in size and shape to the conidia. A melanin-binding monoclonal antibody (mAb) labelled pigmented conidia, yeast cells and the isolated particles as determined by immunofluorescence microscopy. Electron spin resonance spectroscopy revealed that particles derived from pigmented conidia contained a stable free radical compound, consistent with their identification as melanins. Skin tissue from penicilliosis marneffei patients contained yeast cells that were labelled by melanin-binding mAb. Additionally, sera from P. marneffei-infected mice developed a significant antibody response (both IgG and IgM) against melanin. Phenoloxidase activity capable of synthesizing melanin from L-DOPA was detected in cytoplasmic yeast cell extracts. These findings indicate that P. marneffei conidia and yeast cells can produce melanin or melanin-like compounds in vitro and that the yeast cells can synthesize pigment in vivo. Accordingly this pigment may play some role in the virulence of P. marneffei.


Abbreviations: DHN, dihydroxynaphthalene; DOPA, 3,4-dihydroxyphenylalanine; ESR, electron spin resonance spectroscopy; SEM, scanning electron microscopy; TEM, transmission electron microscopy




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