Effect of mixed infections of Sindbis and La Crosse viruses on replication of each virus in vitro

Mixed viral infections are ubiquitous in natural populations and may have significant but unpredictable biological and epidemiological consequences. These infections may be acquired simultaneously (coinfection) or result from two single infections at different time intervals (superinfection). This study investigated the effect of mixed infections of two mosquito-borne viruses, Sindbis (SINV) and La Crosse (LACV), on the replication of each virus in vertebrate baby hamster kidney (BHK) and invertebrate Aedes albopictus (C6/36) cell lines. SINV and LACV were inoculated onto confluent monolayers of each cell type either singly, simultaneously, or with a 2-h delay between single inoculations and titers for each virus were quantified by qRT-PCR. The titers for both viruses were significantly higher in BHK compared to C6/36 cells. Mixed infections suppressed replication of both viruses in BHK cells except for one superinfection treatment where inoculation of SINV ahead of LACV did not result in a reduction in SINV titer relative to single infection treatment. In C6/36 cells mixed infections had no effect on LACV replication, while coinfection enhanced SINV replication. These results demonstrate the potential for mixed viral infections to modify arbovirus transmission and pathogenesis.