Arenaviruses: Cellular Response to Long-Term In Vitro Infection with Parana and Lymphocytic Choriomeningitis Viruses

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Persistent infections were established in suspension cultures of BHK21/13S cells with both Parana and lymphocytic choriomeningitis viruses. Four generations after infection with either virus, more than 90% of the cells scored as infective centers, with concomitant peaks in extracellular virus yields. In both cultures the synthesis of detectable plaque-forming units (PFU) ceased about the 50th generation postinfection, and this condition was maintained until the 350th cell generation when the cultures were discontinued. The generation time of each culture was identical to that of uninfected parent controls, and at no time were cytopathic effects evident. In spite of the absence of infectivity, over 90% of the cells sampled at various times contained viral antigen demonstrable by immunofluorescence. When either of these persistently infected cell lines was substituted for normal cells in the standard plaque assay, very low efficiencies of plating were observed for homotypic and heterotypic viruses. Plaque formation by several heterologous viruses was virtually unaffected. The mechanism of homotypic plaque exclusion in both cell lines was shown to occur beyond the virion adsorption stage. The original infecting virus genome persisted in both cell lines after standard virus was no longer detectable. This was shown with the lymphocytic choriomeningitis virus-infected cells after storage in liquid nitrogen. After thawing, such cells were found to synthesize standard virus for a brief period. Although the Parana virus-infected cells did not behave this way, the growth medium from these cells would initiate PFU synthesis in normal cells within 36 hr after infection.

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