THE CULTIVATION OF DENGUE-1 (HAWAIIAN) VIRUS IN TISSUE CULTURE

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Abstract
After continuous propagation of a dengue-1 virus carrier culture in human skin cells the virus acquired the property of cytopathogenicity (CPE) for human skin (strain HuS 2806) and primary monkey kidney cell cultures. This property persisted in serial passages of the virus in either type of cell culture but cell destruction was more rapid and complete in human skin cell cultures. Peak titers were obtained when CPE appeared 104 mouse LD50 about 78 hours after infection of human skin cell cultures and 105 mouse LD50 about 125 hours after infection of monkey kidney cell cultures. Tissue culture dose (TC CPD50) and mouse LD50 were equivalent when corrected for unit volume. A minimum of 10 hours contact between cells and virus inoculum was necessary in order to obtain satisfactory CPE endpoints. Specific serum neutralization of 100 TC CPD50 was obtained in both cell culture systems with rabbit antisera having the same neutralization index in mice. Staining of infected cultures with fluorescent anti- body (indirect method) resulted in specific fluorescent foci that were indistinguishable from those seen in the cytoplasm of carrier culture cells.