Análise da interação in vitro entre Fonsecaea pedrosoi e macrofagos peritoneais de camundongos C57/BL6 e BALB/c

A cromoblastomicose e uma infeccao subcutanea cronica, granulomatosa, causada pela implantacao traumatica de diversas especies de fungos demaceos, sendo Fonsecaea pedrosoi o principal agente etiologico. O Brasil possui a segunda maior prevalencia mundial da doenca, sendo o estado do Para a maior area endemica. Histologicamente, a cromomicose e caracterizada pela presenca de celulas gigantes, onde podem ser observadas celulas escleroticas fagocitadas por macrofagos. O objetivo do presente estudo foi analisar os diferentes aspectos da interacao de macrofagos peritoneais de camundongos BALB/c e C57/BL6 com conidios ou celulas escleroticas de F. pedrosoi, determinando os indices de infeccao, fagocitose e fusao celular. Os resultados mostraram indices de fagocitose e infeccao maiores em conidios do que em celulas escleroticas para BALB/c (p<0.05), ocorrendo efeito inverso no indice de fusao, com a formacao de celulas gigantes do tipo Langhans na interacao com celulas escleroticas e celulas gigantes do tipo corpo estranho na interacao com conidios. Os macrofagos de BALB/c em interacao com conidios produziram mais TNF-α que o controle nos tempos de 3 a 72h; e mais IL-10 apos 3h. Macrofagos interagindo com celulas escleroticas produziram mais TNF-α que o controle nos tempos de 1h e 3h; e a quantidade de IL- 10 foi maior apos 72h de interacao. No co-cultivo de macrofagos de C57/BL6 com conidios observou-se a presenca de vacuolos aumentados apos 24h, enquanto na interacao com celulas escleroticas, os macrofagos se desprenderam da laminula nos tempos posteriores a 24 h. A quantidade de TNF-α e maior na interacao de conidios comparado ao controle em 1 e 72 h; e a quantidade de IL-10, no tempo de 48h. Ja na interacao com celulas escleroticas, apenas a quantidade de IL-10 diferiu do controle, sendo maior nos tempos de 1 a 48h. Estes dados sugerem que a resposta e macrofagos ao fungo e diferente entre os camundongos de BALB/c e C57/BL6, diferindo tambem a resposta de um mesmo tipo de macrofago para cada forma fungica, sendo as celulas escleroticas aparentemente mais imunogenicas que os conidios.

ABSTRACT: Chromoblastomycosis (CBM) is a chronic, subcutaneous, granulomatous infection caused by traumatic implantation in the skin of several dematiaceous fungi, usually Fonsecaea pedrosoi. Brazil has second highest disease prevalence in the world and Para State is the most endemic area. Histologically, CBM is characterized by the presence of multinucleated giant cells and sclerotic cells can be found engulfed by macrophages. The objective of this study was to analyze the different aspects of interaction between peritoneal macrophages from BALB/c or C57/BL6 mice with F. pedrosoi conidia or sclerotic cells, calculating infection, phagocytosis and cellular fusion rates. The results showed phagocytosis and infection rates with conidia higher than sclerotic cells to BALB/c (p <0.05), while the rate of cellular fusion was higher for sclerotic cells interaction, with Langhans giant cells formation, in comparison to foreign-body giant cells after interaction with conidia. Macrophages from BALB/c co-cultured with conidia produced more TNF-α than control group after 3 to 72 hours, and more IL-10 after 3h. Macrophages interacting with sclerotic cells produced more TNF-α than control group after 1h and 3h, and the amount of IL-10 was higher after 72h of interaction. In the co-culture of C57/BL6 macrophages with conidia, the presence of large vacuoles after 24h was observed, while in the coculture with sclerotic cells, macrophages were detached from coverslip glasses after 24 h. Our results indicate higher levels of TNF-α after interaction of conidia compared to controls at 1 and 72 h and increase of IL-10 after 48h. However, after interaction with sclerotic cells, only IL-10 differed from control, being higher after 1 to 48 hours. All of these data suggest that macrophage response to fungus is different between BALB/c and C57/BL6 mice, differing also on the response of the same type macrophage for each fungal form, sclerotic cells apparently being more immunogenic than conidia.