Vainikolo, who qualifies for England on residency grounds, signed a three-year contract with Guinness Premiership leaders Gloucester last summer.
He is the Premiership's top try scorer this season, and possesses explosive finishing skills.
But Vainikolo's selection will court controversy, especially as players such as Josh Lewsey and James Simpson-Daniel have been omitted.
Vainikolo's fellow rugby league recruit Andy Farrell misses out, and there is also no place for Leicester centre Dan Hipkiss.
The uncapped players are Vainikolo, Wasps fly-half Danny Cipriani, Sale scrum-half Richard Wigglesworth and Leicester flanker Tom Croft.
England World Cup captain Phil Vickery is retained as skipper.
England 32-man squad for RBS 6 Nations Championship
Backs: I Balshaw (Gloucester), D Cipriani (Wasps), M Cueto (Sale Sharks), T Flood (Newcastle), A Gomarsall (Harlequins), C Hodgson (Sale Sharks), J Noon (Newcastle), P Richards (London Irish), P Sackey (Wasps), D Strettle (Harlequins), M Tait (Newcastle), M Tindall (Gloucester), R Wigglesworth (Sale Sharks), J Wilkinson (Newcastle), L Vainikolo (Gloucester).
Forwards: S Borthwick (Bath), T Croft (Leicester), G Chuter (Leicester), L Deacon (Leicester), N Easter (Harlequins), J Haskell (Wasps), B Kay (Leicester), L Mears (Bath), L Moody (Leicester), T Payne (Wasps), T Rees (Wasps), M Regan (Bristol), S Shaw (Wasps), A Sheridan (Sale Sharks), M Stevens (Bath), P Vickery (Wasps, capt), J Worsley (Wasps).