Human peripheral blood polymorphonuclear leucocytes (PMN) were examined in vitro for antifungal activity against yeast cells of Paracoccidioides brasiliensis. The yeast cell of this fungus was resistant to killing by PMN. However, PMN exhibited a fungistatic effect on the fungal isolates employed in a long-term ( approximately 72 h) assay. Lysates of PMN did not show a fungistatic or fungicidal effect, indicating that live PMN are necessary for the antifungal effect. Interferon-Gamma (IFN-Gamma) enhanced the antifungal activity of PMN. IFN-Gamma-treated PMN killed isolate Bt-4 within 2 h of coculture, and after 24 h a still greater killing effect was observed. By contrast, IFN-Gamma-treated PMN did not show a significant killing effect on isolate Tatu, but did exhibit an enhanced fungistatic effect on this isolate. In contrast, tumor necrosis factor-alpha and interleukin 8 had no effect on the antifungal activity of PMN. Scanning electron microscopy revealed that the surface structure of the fungal cell was apparently damaged by PMN within 24 h of cocultures. Based on these novel findings, we speculate that human PMN might play a role in host resistance in early infection with this fungus due to their antifungal activity.