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Exploratory experiments were performed with temperature-sensitive (ts) mutants of Sindbis virus for studies on viral pathogenesis in the embryonated egg, a host which is immunologically underdeveloped. Parent and mutants were found to be virulent at the permissive temperature (33 C), but only the mutants were attenuated at the nonpermissive temperature (38.5 C). The degree of attenuation varied with the mutant and the route of inoculation. Chicks which survived infection by ts mutants at a nonpermissive temperature weighed the same as controls and showed no obvious abnormalities on gross examination. Whenever death of the embryo at the nonpermissive temperature occurred after inoculation with a mutant, it was apparently due to the selection of a population of temperature-insensitive virulent revertants. Kinetic studies showed that, after inoculation of the chorioallantoic membrane and incubation at the permissive temperature, a number of cycles of virus multiplication and dissemination apparently occurred rapidly. At the nonpermissive temperature, multiplication was undetectable. Certain pathophysiological signs were seen in the slower, less virulent infections by the mutants at the permissive temperature that were apparently masked or obscured in the more virulent, rapid infection by the parent. From these results and those reported in a subsequent paper, it appears that ts mutants of viruses possess potential as valuable tools for analyzing pathogenesis and immunity in the intact animal host that are complementary to more conventional approaches which employ normal (temperature-insensitive) viruses.

Pathogenesis of Temperature-Sensitive Mutants of Sindbis Virus in the Embryonated Egg I. Characterization and Kinetics of Viral Multiplication