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Reports on the arrest of herpes simplex virus type 1 (HSV-1) replication by interferon (IFN) are inconsistent. By the use of immunofluorescence and immunoblot assays with monoclonal and polyclonal antibodies, effective arrest of viral translation by human IFN-α in human fibroblasts was detected for the HSV-1 strains KOS and McIntyre. In HeLa cells which are less sensitive to IFN inhibition and in 444 cells, a HeLa-fibroblast hybrid cell line, the inhibition was less pronounced. These results confirm earlier observations that IFN or polyinosinic-polycytidylic acid block the replication of HSV-1 in human, monkey and mouse cells no later than the immediate early phase of infection.