Membrane-associated enteroviruses undergo intercellular transmission as pools of sibling viral genomes

Some viruses are released from cells as pools of membrane-associated virions. By increasing the multiplicity of infection (MOI), this type of collective dispersal could favor viral cooperation, but also the emergence of cheater-like viruses such as defective interfering particles. To better understand this process, we examined the genetic diversity of membrane-associated coxsackievirus infectious units. We find that infected cells release membranous structures (including vesicles) that contain 8–21 infectious particles on average. However, in most cases (62%–93%), these structures do not promote the co-transmission of different viral genetic variants present in a cell. Furthermore, collective dispersal has no effect on viral population sequence diversity. Our results indicate that membrane-associated collective infectious units typically contain viral particles derived from the same parental genome. Hence, if cooperation occurs, it should probably involve sibling viral particles rather than different variants. As shown by social evolution theory, cooperation among siblings should be robust against cheater invasion.R.S. was funded by ERC Consolidator Grant 724519 Vis-a-Vis. J.-V.B. was funded by a Prometeo Grant 2016/122 from the Generalitat Valenciana. R.G. was funded by the Spanish Ramón y Cajal program (RYC-2015-17517) and Ministerio de Ciencia e Innovación (grant BFU2017-86094-R).Peer reviewe