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Establishment of Herpes Simplex Virus Latency in vitro with Cycloheximide

Department of Microbiology, The Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033, U.S.A.

Human embryonic lung cells were infected with herpes simplex virus (HSV), treated with 10 µg/ml or more of cycloheximide for 24 h, incubated at 37 °C, and then shifted to 40.5 °C for various periods of time (0 to 40 days) without cycloheximide treatment. No infectious virus was detected after freezing and thawing of the cultures; however, infectious virus was recovered after temperature shift-down to 37 °C or superinfection with human cytomegalovirus (HCMV). The time course for formation of infectious centres after temperature shift-down was examined with and without HCMV superinfection during incubation at 40.5 °C. Two patterns of latently infected cells were identified: one pattern showed spontaneous reactivation of virus after temperature shift-down, and the second showed reactivation of HSV after superinfection with HCMV. The first pattern showed a rapid decrease in the number of infectious centres with time, whereas the second maintained a steady reactivation rate up to 40 days at 40.5 °C. The same tendency was observed for infectious centre formation at 37 °C with and without HCMV superinfection in the HSV latency system established with (E)-5-(2-bromovinyl)-2'-deoxyuridine and interferon treatment.

Keywords: HSV, latency, cycloheximide

Received 12 May 1986; accepted 7 August 1986.

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