Giardiavirus Enters Giardia lamblia WB Trophozoite via Endocytosis

Abstract

As part of our continuing effort in the study of Giardiavirus life cycle, the early events of Giardia lamblia virus (GLV) infection in the trophozoites of G. lamblia WB strain were examined. Electron microscopy showed that GLV particles were initially localized on plasma membrane. As time progresses, GLV was translocated to the peripheral vacuole and then spread to the cytoplasm. Inhibitors of endocytosis such as sodium azide, chloroquine, or ammonium chloride disrupted viral infection when the drug was added simultaneously with the infecting GLV. The inhibitory effect was reduced when sodium azide or chloroquine was added at various intervals after infection. When cells infected for 1 hr were examined by immunofluorescence staining, sodium azide greatly reduced GLV staining signal in general while chloroquine restricted the staining signal to a few bright spots. Significantly more GLV was found in the peripheral vacuoles of chloroquine-treated cells than untreated controls by semiquantitative electron microscopy. In contrast, only a reduced amount of GLV was found in the peripheral vacuoles of sodium azide-treated cells. These results suggest that sodium azide reduces the internalization of infecting GLV, while chloroquine confines the virus in the peripheral vacuoles and, consequently, leads to nonproductive infection. We conclude from these observations that the entry of GLV into the susceptible WB cells in the event of infection is most likely mediated by endocytosis.