Growth characteristics of poliomyelitis virus in HeLa cell cultures; lack of parallelism in cellular injury and virus increase

Abstract

A detailed study of the cytological changes which are induced in HeLa cells by the Saukett strain of Type III poliomyelitis virus has been made. The observations were of cultures in which a single sequence of infection was induced. The cytological changes were examined in relation to the growth curve of the virus in the same type of culture. This curve showed a latent period of 4 to 5 hours, followed by a gradual release of virus over an interval of 6 to 7 hours. Changes in the staining character of the cells occurred before the major portion of the viral yield appeared. The infected cells exhibited a striking cytopathology with increased basophilia, nuclear pyknosis, and basophilic cytoplasmic granules. Individual cells showed characteristic differences in the rate at which the cytopathology progresses. The multiplication of the virus in HeLa cells was inhibited by fluorophenylalanine. The inhibitory effectiveness of the antimetabolite was related to the age of the infection. It apparently inhibits only an early stage of viral development. The inhibition is completely reversed by phenylalanine if the amino acid is added within 6 hours, not later, after the induction of virostasis. The data are interpreted in terms of the rate at which the ability of the infected cell to support viral synthesis was lost. Flurophenylalanine also inhibited the multiplication of HeLa cells; however, the effect upon the uninfected cell was reversible after 3 days, as indicated by viability after such treatment. While the fluoro derivative completely inhibited viral multiplication, it did not prevent the cytopathogenic effect of the virus. In the presence of fluorophenylalanine, the disintegration of an infected cell proceeded at what appeared to be the ordinary rate, without any increase of the infectious agent. Experimentally the processes leading to viral increase and to cellular injury have been shown to possess a significant degree of autonomy.