Distribution of glucose and mannose moieties of Acanthamoeba sp. (W4) Philippine isolate was detected using fluorescem isothiocyanate (FITC)- labeled Concannavalin A (Con A) lectin. Green fluorescent patches around the plasma membrane of agglutinated trophozoites (Fig.1) were observed. Isolated Acanthamoeba cyst exhibited strong fluorescence on the cyst wall Brighter fluorescence was detected on the site of adherence between the Acanthamoeba (W4) cysts and trophozoites (Fig. 1,3). These lectin receptors were concentrated at the uroidal region of the trophozoite. The fluorescence, however, was absent in the newly forming hyaline cap (Fig.4). Upon addition of α -methyl-mannoside (0.5 M), Con A binding to sugar moieties in cyst and trophozoites was blocked and no fluorescence was observed.
The binding specificity of Con A-FITC and Acanthamoeba cell surface mannose moieties demonstrate topographical distribution of lectin receptor sites. Ultrastructurally, ferritin-labeled Con A at cell adhesion sites showed clustering of lectin receptors. Occurrence of fluorescence in Naegleria sp. using Con A-FITC has been shown to concentrate at the uroidal region but no fluorescence was seen at the anterior of newly formed pseudopodia.