HeLa cells infected with several group B coxsackieviruses contain a previously undetected, virus-specific ribonucleoprotein particle which we designated membrane-bound virion (MBV). MBVs of B5 virus have a pronounced polygonal appearance and are slightly smaller than virions. The particles sediment more slowly (at about 107S) and have a lower buoyant density (about 1.30). They contain 35S virion RNA; only three, and not four, capsid proteins; and at least seven additional proteins with apparent molecular weights of 21,000 to 92,000. Three of the latter proteins appear to be of host origin; the rest may be precursors of virion capsid proteins. The RNA is resistant to digestion by RNase, and EDTA treatment disrupts the particle. MBVs are infectious, although significantly less so than virions. Cells infected with MBVs produce both types of progeny, virions and MBVs. In coinfected cultures, the yield of progeny is lower than in cells infected with virions alone, suggesting interference by MBVs. Synthesis of both types can be detected within 3.5 h after infection, and synthesis continues for 24 h.