TGF-beta plasma levels in chromoblastomycosis patients during itraconazole treatment.

BACKGROUND: Chromoblastomycosis (CBM) is a dermal mycosis. The disease evolves to a chronic state, presenting a suppurative granulomatous dermatitis, combined with variable dermal fibrosis. Pathogenesis of the inflammation and tissue repair in CBM are poorly understood. AIM: To quantify Transforming Growth Factor-beta (TGF-beta) plasma levels of CBM patients during itraconazole (ITZ) treatment. METHODS: Blood plasma of 12 CBM patients was subjected to TGF-beta titration with ELISA at 0, 3, 6 and 12months of 200mg per day of ITZ therapy, and correlated with the clinical aspects. Plasma of 12 healthy individuals were used for control. RESULTS: CBM patients present high plasma levels of TGF-beta (7.016+/-1988pg/ml), decreasing after 03months (4.625+/-645pg/ml) of ITZ treatment, which correlates with a rapid clinical improvement. However, after 6 (6.566+/-777pg/ml) and 12months (6.908+/-776) of treatment, TGF-beta levels increase to almost the same levels observed before treatment, which is related to a slow clinical improvement, fungal persistence on the lesion, and fibrotic scars. CONCLUSION: TGF-beta plasma levels are high in CBM patients. Fungal destruction by ITZ correlates with TGF-beta downregulation, but tissue remodeling and fungal persistence probably raises its levels again, interfering with cellular immune responses.

Enzymatic isolation of Lacazia loboi cells from skin lesions of lobomycosis.

Lacazia loboi is the etiologic agent of Jorge Lobo's disease, a cutaneous and subcutaneous mycosis endemic to Latin America tropical regions and characterized by chronic nodular or keloidal lesions which develop after traumatic events. A new method for the extraction of L. loboi yeast cells from biopsies of lobomycosis skin lesions is presented. The method is based on the proteolytic action of the enzyme dispase which is known for its action against fibronectin and collagen type IV. Fungal identification was based on histological examination of the biological material and molecular analysis based on 18S ribosomal sequences. Observations under optic and fluorescence microscopy proved the efficacy of enzymatic isolation of the lobomycosis etiologic agent, as well as identifying the organism's main parasitic characteristics. Molecular phylogenetic analysis corroborated the histological examination and indicated L. loboi relationship with other members of the Onygenales. Use of dispase proved to be ideal for the isolation of L. loboi from human biopsies, shows promise as an important tool for improving biological studies of this peculiar fungus.

Development of natural culture media for rapid induction of Fonsecaea pedrosoi sclerotic cells in vitro.

Fonsecaea pedrosoi is the main agent of chromoblastomycosis, a skin disease presenting verrucous lesions, in which round, thick-walled sclerotic cells are found. In vitro induction of sclerotic cells is time-consuming (20 to 45 days) and temperature dependent. We present two new natural media that reduce the sclerotic-cell induction time to only 2 days.

Phagocytosis of Fonsecaea pedrosoi conidia, but not sclerotic cells caused by Langerhans cells, inhibits CD40 and B7-2 expression.

Fonsecaea pedrosoi is the major etiological agent of chromoblastomycosis, a chronic, suppurative, granulomatous mycosis usually confined to skin and subcutaneous tissues, presenting a worldwide distribution. The host defense mechanisms in chromoblastomycosis have not been extensively investigated. Langerhans cells (LC) are bone-marrow-derived, dendritic antigen-presenting cells of the epidermis, which constitutively express major histocompatibility complex (MHC) class II, and comprise 1-3% of total epidermal cells. LC are localized in suprabasal layers of the epidermis and in mucosa, where they play important roles in skin immune responses. The purpose of the present study was to evaluate the interaction of F. pedrosoi conidia or sclerotic cells with LC purified from BALB/c mice skin. We demonstrate here that LC phagocytose F. pedrosoi conidia but not sclerotic cells in the first 3 h of interaction, inhibiting hyphae formation during 12-hour coculture from both forms, internalized or not. Also, LC maturation, analyzed using CD40 and B7-2 expression, was inhibited by conidia, but not by sclerotic cells, indicating an important innate immunity function of LC against F. pedrosoi infection in these mice.

Isolation of Fonsecaea pedrosoi from thorns of Mimosa pudica, a probable natural source of chromoblastomycosis.

We report the isolation of Fonsecaea pedrosoi from thorns of the plant Mimosa pudica L. at the place of infection identified by one of our patients. Clinical diagnosis of chromoblastomycosis was established by direct microscopic examination and cultures from the patient's lesion. The same species was isolated from the patient and from the plant. Scanning electron microscopy of the surface of the thorns showed the characteristic conidial arrangement of F. pedrosoi. These data indicate that M. pudica could be a natural source of infection for the fungus F. pedrosoi.