Fc receptors on the surface of Toxoplasma gondii trophozoites: A confounding factor in testing for anti-Toxoplasma antibodies by indirect immunofluorescence

Abstract

Negative sera often produce false-positive polar patterns of fluorescence in indirect immunofluorescence tests for serum anti-Toxoplasma gondii antibodies, representing a confounding factor in the diagnosis of toxoplasmosis. In this work, we studied the T. gondii trophozoites, the antigenic material used in the immunofluorescence tests, expressed surface Fc receptors that could cause the binding of normal immunoglobulins, thus producing false-positive results. We report here that T. gondii trophozoites indeed have Fc receptors on their surface. This was shown by the direct binding of purified human Fc to the parasite, evidenced by the subsequent binding of fluorescein-labeled Fab specific for human Fc. In addition, pretreatment of the parasite with excess purified Fc to saturate the surface Fc-binding sites abrogated the formation of polar fluorescence. The trophozoites appeared to express Fc receptors with different degrees of affinity for the specific ligand, since a diffuse fluorescence pattern was observed following incubation with 1 mg of Fc per ml (10 μg per well), whereas with 0.2 mg (0.2 μg per well), a majority of parasites showed polar fluorescence. This observation suggests that the Fc receptors accumulated at the polar cap are those with higher affinity. The present findings raise intriguing questions regarding the possible biological role(s) of the Fc receptors on T. gondii but, more immediately, indicated that pretreatment of the antigenic material with Fc, a commercially available reagent, constitutes a practical, simple way to avoid false-positive immunoflourescence test results due to the binding of nonspecific immunoglobulin to the parasite.