The expression of herpes simplex virus type 2 (HSV-2) glycoproteins on the surface of human neuroblastoma cells has been investigated using Millipore Millicell culture plate inserts. Utilizing a modified radioimmunoassay, we learned that glycoproteins B, C, D, E, and I were expressed predominantly on the apical membrane domain of the infected neuroblastoma cells. The unidirectional transport of HSV-2 glycoproteins was substantiated by the analysis of extracellular glycoproteins released from neuroblastoma cells. The results suggest that the evaluated HSV-2 glycoproteins were transported primarily to the apical plasma membrane domain of human neuroblastoma cells. © 1991.