Identification, Characterization, and In Vitro Culture of Highly Divergent Arenaviruses from Boa Constrictors and Annulated Tree Boas: Candidate Etiological Agents for Snake Inclusion Body Disease

Stenglein, Mark D.; Sanders, Chris; Kistler, Amy L.; Ruby, J. Graham; Franco, Jessica Y.; Reavill, Drury R.; Dunker, Freeland; DeRisi, Joseph L.

Hypothesizing that boa constrictor cells might be permissive for replication of the boa constrictor-derived virus, we harvested tissue from a 22-year-old female boa constrictor ("Juliet") who died of lymphoma and attempted to create continuous cell lines (39) . Of the multiple tissues and conditions attempted, proliferation was evident only in a culture of adherent cells derived from Juliet's kidneys (Fig. 4, inset) . This cell line (JK cells) continues to proliferate, although it is not yet clear whether it is immortal. We inoculated a culture of JK cells with liver and kidney homogenates from the CAS boa constrictors and observed an exponential increase of viral RNA in culture supernatant after an initial decrease (Fig. 4) . Viral RNA was not detectable in mock-inoculated JK cultures. Infected cells did not exhibit gross cytopathic effects over the course of 21 days.