In Vitro Studies on Rickettsia-Host Cell Interactions: Intracellular Growth Cycle of Virulent and Attenuated Rickettsia prowazeki in Chicken Embryo Cells in Slide Chamber Cultures

Abstract

Upon entry into the cytoplasm of irradiated chicken embryo cells in slide chamber cultures infected over a 2-h period, yolk sac-grown virulent (Breinl strain) and attenuated (E strain) Rickettsia prowazeki underwent indistinguishable reproducible intracellular growth cycles. They promptly entered an exponential growth phase, without detectable lag and without microscopic evidence for any unusual early replicative phase. The generation time for both strains was 8.8 to 8.9 h at 34 C. During most of this period, the state of the organisms and growth were very similar from one cell to another. The exponential-growth phase continued for at least 36 to 48 h, when the rickettsiae became too numerous to count by microscopic examination. Between about 36 and 48 h, cells packed with rickettsiae began irregularly to break down and release organisms. These began to initiate new infection cycles in previously uninfected cells over many hours, as demonstrated by the rise in percentage of cells infected, yielding a highly disordered infected culture with different cells containing rickettsiae in diverse stages of growth. The organisms underwent regular minor changes in morphology, similar to those seen in bacterial cultures, in the first infection cycle. As the cells became packed with rickettsiae, the microorganisms regularly diminished in size to become minute coccobacillary to coccoid forms. However, the rickettsiae in the second and subsequent infection cycles in aging cultures often assumed filamentous or swollen bizarre forms. Only the first infection cycle conformed closely to the concept of a one-step growth cycle. A set of terms is proposed and defined for the infection cycle.