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Melanin has been proposed as a virulence factor inCryptococcus neoformans , but its presence has not been shown unambiguously in vivo. Validated methods used previously to show production of cryptococcal eumelanin pigment in vitro (P. R. Williamson, K. Wakamatsu, and S. Ito, J. Bacteriol. 180:1570–1572, 1998) were used to assess for production of laccase-derived products in mouse brain of the Lacc+ strains, 2E-TUC, H99 (serotype A), and ATCC 34873 (serotype D), and the Lacc− strain, 2E-TU. Pyrrole-2,3,5-tricarboxylic and pyrrole-2,3-dicarboxylic acid, specific degradation products of catecholamine derivatives such as melanin, were found in all Lacc+ strains, but not in the Lacc− strain, 2E-TU. However, the presence of melanin pigment itself could not be demonstrated in the same cells. Lack of the specific degradation products aminohydroxyphenylalanine and aminohydroxyphenylethylamine in Lacc+ strains upon hydriodic acid hydrolysis showed that pheomelanin was also not produced by the fungus in vivo. These are the first data to support the generation of catecholamine oxidation products byC. neoformans in vivo, but they do not support postenzymatic polymerization of these products to form typical eumelanin, as previously proposed.

Catecholamine Oxidative Products, but Not Melanin, Are Produced byCryptococcus neoformansduring Neuropathogenesis in Mice