The effect of opsonization of Pneumocystis carinii with different antibody classes, complement, mannan-binding lectin (MBL), and lung surfactant protein D (SP-D) on respiratory burst activation was studied. Antibodies were obtained by affinity chromatography, complement from a hypogammaglobulinaemic patient, and phagocytic cells from blood donors. Respiratory burst activation was measured by chemiluminescence (CL). With freshly isolated neutrophils the combination of antibodies and complement but not antibody alone, had opsonizing properties. With neutrophils cultured for 20 h, however, IgG increased the CL response. In macrophages P. carinii opsonized with IgG alone induced a CL response proportional to the antibody titre used. With IgA an effect, albeit lower, was also seen, whereas IgM alone was inefficient. The combined effect of antibodies and complement increased the response significantly for all three antibody classes, IgG and complement giving the largest response. Binding of MBL to P. carinii and Candida albicans was demonstrated; however, only the former stimulated activation of the respiratory burst. SP-D did not bind to either microorganism and had no effect on the respiratory burst. It is concluded that IgG, IgA and complement are important opsonizing factors in infections involving P. carinii. The relative importance varies with the type of phagocytic cell studied.