Rhabdomyosarcoma monolayers were inoculated with enterovirus 71 (EV 71) at 73 degrees C, sampled at intervals during the replicative cycle, and examined in thin sections by electron microscopy, using routine and immunoelectronmicroscopy with polyclonal antibodies against EV 71. The location of EV 71 or its precursors was followed during the viral replicative cycle. The earliest samples (3 h postinoculation) showed a cell shape change, from elongated to rounded. At 6 h postinoculation, the presence of early virus-induced vesicles developing within the cytoplasm was pointed out, although no virus particles were observed at these stages. At 12 and 20 h postinoculation, virus particles appeared in the cytoplasm. They were found free or in clusters in the cytoplasmic matrix, between the virus-induced vesicles. EV 71 particles were also occasionally observed in the form of paracrystalline arrays situated in the vesiculated areas. The immunolabel (gold beads) was found, initially, over dense cytoplasmic areas and in more advanced process at the vesiculated area and over the virus particles. Therefore the main cellular alterations observed in this infection were the shape change of the cell and the appearance of electron-dense areas (viroplasm) and smooth walled vesicles which are probably the site of the virus replication.