Observations of Aedes albopictus cell cultures persistently infected with Sindbis virus.
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Biomedical subjects
Publications and source records attributed to V Stollar.
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The 5' sequence of Sindbis viral RNA is m (7)G(5') pppApUpGp...
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Cultures of Aedes albopictus cells persistently infected with wild-type Sindbis virus (SV-W) give rise to small plaque-forming mutants which are also temperature sensitive. These mutants, designated SV-C, are neutralized by antiserum produced against SV-W. Mutant ts clones were isolated from SV-C by plaque purification. After serial undiluted passage in BHK or mosquito cells, each of the clones gave rise to ts(+) revertants which, however, remained mutant with respect to plaque morphology. Nineteen of 20 clones derived from SV-C were RNA(+), and one was RNA(-) (SV-C-2). The RNA synthesizing activity, once induced in infected cells by SV-C-2, was stable at the nonpermissive temperature (39.5 C). All clones derived from SV-C were inactivated at 60 C much more quickly than was SV-W. It was not possible to demonstrate complementation between any of the SV-C clones.
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Complete homologous interference is demonstrated in cultures of Aedes albopictus cells chronically infected with Sindbis virus. The interference occurred before there was any detectable RNA synthesis by the superinfecting virus.
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A quantitative complement fixation assay which specifically measures double-stranded RNA has been used to study this RNA extracted from uninfected and arbovirus-infected cells. The double-straned RNA of the uninfected BHK-21 cells sedimented in the 12S region in sucrose gradients. The double-stranded RNA of Sinbis virus-infected cells, as measured immunochemically, included a predominant peak at 12S, a smaller 18S peak, and polydisperse material extending into the 26-30S region. All classes of this RNA detected immunochemically showed a sharp thermal denaturation curve, and increased in amount progressively during infection, with the 12S peak predominant at all times.
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