Fluorescent Antibody Studies with Herpes Simplex Virus in Unfixed Preparations of Trypsinized Tissue Cultures

Summary: The classical strain of herpes simplex virus (Burnet & Lush, 1939) and a recently isolated strain (Melbourne 1/56) were propagated in tissue culture of human amnion and infant-mouse kidney ‘epithelial’ cells. Following the inoculation of 103·25 TCCD50 (tissue culture cytopathogenic dose) of the Melbourne 1/56 herpes simplex virus into amnion cell culture, the titre of virus dropped to 100·50 TCCD50 at 12 hr. and the maximum level of virus (103·50 TCCD50) was detected at 48 hr. after inoculation. The earliest morphological changes were observed in the tube-cultures c. 18 hr. after inoculation of the virus and eosinophilic intranuclear inclusion bodies were observed in the amnion cells c. 24 hr. after virus injection. Cell degeneration was prominent at 36 hr. when most cells were highly retractile and rounded. Cell suspensions, harvested from infected and uninfected cultures of amnion and mouse- kidney epithelium, were treated with immune herpes serum followed by fluorescent rabbit anti-human γ-globulin, and specific peripheral fluorescence was observed in the unfixed preparations of the infected tissue cultures. The limited surface staining was ascribed to the impermeability of the unfixed cytoplasmic membranes to globulins.