Table 1. Mutations detected in examples of viruses showing large plaques and/or loss of RSV F expression.
Virus clone | Construct | Plaque size | RSV F expression detected | RSV F Mutation* | HPIV3 Vector mutation** |
---|---|---|---|---|---|
D3-F12 | F/preN | Normal | no | Y342C | ND |
D11-H5 | F/preN | Mixed- normal and large | no | E110 → Stop | None found |
E1-B4 | F/preN | Normal | no | E110 → Stop | ND |
G9-E10 | F/preN | Normal | no | “A” insertion at nt 5 caused frame shift and E30 → Stop | ND |
C5-E3 | F-H3TMCT/preN | Large | yes | No | Y3H (HN protein); V242G, N248K, C252W, F277V, I279M (L protein) |
A3-D8 | F/N-P | Normal | no | None found | ND |
C6-F4 | F/N-P | Large | no | L321P, Y342H, F352P | None found |
G6-E8 | F-H3TMCT/N-P | Normal | no | K132 → Stop | ND |
One stock of each virus construct, at P1 stage after rescue, was subjected to terminal dilutions in LLC-MK2 cells. Terminally diluted enriched virus clones that showed large plaque phenotype and/or had lost expression of RSV F were sequenced.
* RSV F insert and flanking transcription signals were sequenced for viruses with loss of RSV F expression.
** HPIV3 backbone was sequenced for clones with large plaques.
ND, not determined.