The human epidermal cells infected with herpes simplex virus in vitro

Summary

The cytopathic changes of the epidermal cells infected with herpes simplex virus were observed in vitro. Cell cultures were initiated from suspension of pure epidermal cells obtained by tryptic digestion of the adult human skin. The keratinocytes connected by intercellular bridges were to form confluent sheets. The melanocytes were scattered around the sheet and often stretched dendritic processes towards the keratinocytes.

Herpes simplex viruses were inoculated to the culture. The cytopathic effects on the keratinocytes were as follows: The ruffling membrane activity which was usually observed on the keratinocytes along the free margin of the sheet was lost at two hours after the inoculation. At six hours the intercellular connections were lost specifically at the periphery of the sheet and the polygonal keratinocytes began to take round, elliptic or spindle-shaped forms. At 24 h the degeneration of the keratinocytes was prominent. The nuclei showed vacuolated appearance and giant cells were simultaneously observed. The morphological changes of melanocytes, however, were minimal. The titer of the virus in the medium increased as high as 10^9.33 (TCID₅₀/ml) at 50 h after the inoculation.

Zusammenfassung

Die cytopathischen Veränderungen von Herpes simplexinfizierten Epidermiszellen wurden in vitro beobachtet. Kulturen reiner Epidermiszellen wurden nach tryptischer Verdauung menschlicher Erwachsenenhaut angesetzt. Die Keratinocyten waren durch Intercellularbrücken verbunden und bildeten zusammenhängende Zellrasen. Darum verstreut lagen Melanocyten, deren dendritische Fortsätze oft mit den Keratinocyten verbunden waren.

Folgende cytopathischen Veränderungen zeigten sich nach Inoculation der Kulturen mit Herpes simplex-Viren: 2 Std nach Inoculation ging in den am Rande des Zellrasens befindlichen Keratinocyten die Membranbewegungen verloren. Nach 6 Std lösten sich die Intercellularbrücken an der Peripherie. Die polygonalen Keratinocyten nahmen runde, elliptische oder spindelige Formen an. Degeneration der Keratinocyten trat nach 24 Std ein, wobei vacuolierte Kerne wie auch Riesenzellbildung beobachtet werden konnte. An den Melanocyten wurden nur minimale Veränderungen sichtbar. 50 Std nach Inoculation war der Virustiter im Medium auf 10^9.33 (TCID₅₀/ml) angestiegen.