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Propagation of Yellow Fever Virus (17D Strain) in Primary Trypsinized Cell Cultures^*

The 17D strain yellow fever (YF) virus multiplied in primary trypsinized kidney cell cultures from chicken, guinea pig, hamster, hog and Japanese monkey (Macaca fuscata), but apparently not in rabbit kidney cell cultures. The longest periods in which active virus was detected in the infected culture fluid and the highest viral titers (mouse-intracerebral LD₅₀/0.02 ml) obtained in the present study were: chicken kidney, 30 days, 10^3.5; guinea pig kidney, 14 days, 10^3.25; hamster kidney, 36 days, 10^4.25; hog kidney, 50 days, 10^3.75; and monkey kidney, 43 days, 10^3.5.

The 17D virus was transmitted through a number of hamster kidney cell ultures. Infectivity for mice was shown with the 44th passage's culture fluid representing 10^-68 dilution of the starting inoculum. The cultivated virus retained its mouse-infectivity throughout the subcultures.

The infected hamster and monkey kidney cells exhibited degeneration visible under an ordinary light microscope. The cellular degeneration was prevented by antiserum from rabbits immunized with the original 17D virus. The in vitro neutralization paralleled the conventional mouse brain tests. A tendency was shown that ID₅₀ values of 17D virus for hamster kidney cell cultures were higher than LD₅₀'s for 3-week-old mice, and that the cellular degeneration in cultures appeared earlier than the death of mice. Similar results were obtained in experiments using the mouse brain or cell culture passaged virus or the commercial chick embryo vaccine virus.

^* Preliminary accounts for part of this work were presented at meetings of the Society of Japanese Virologists, June 1960, and the Japanese Society of Tropical Medicine, September 1960.