Bukayo Saka doesn’t make the squad after suffering a "slight knock" in training. Mason Mount is recalled after returning from isolation.
Southgate had decisions to make about four players walking a tightrope on yellow cards - Harry Maguire, Declan Rice, Kalvin Phillips and Phil Foden. The first three all start.
Those bookings would expire after this round, but those players risk missing the semi-final if they pick up another yellow at the Stadio Olimpico.
Ukraine are without striker Artem Besedin, who suffered a serious knee injury during a wild challenge from Sweden defender Marcus Danielson that earned the latter a red card in extra time at Hampden Park on Tuesday.
Here is how the two sides shape up...
England vs Ukraine starting lineups
Ukraine (3-5-2): Bushchan; Karavaev, Zabarnyi, Kryvtsov, Matviyenko, Mykolenko; Sydorchuk, Zinchenko, Shaparenko; Yarmolenko, Yaremchuk
England (4-2-3-1): Pickford; Walker, Stones, Maguire, Shaw; Phillips, Rice, Mount; Sancho, Kane, Sterling
Goalkeepers: Sam Johnstone, Jordan Pickford, Aaron Ramsdale*
Defenders:, Ben Chilwell, Conor Coady, Reece James, Harry Maguire, Tyrone Mings, Luke Shaw, John Stones, Kieran Trippier, Kyle Walker, Ben White
Midfielders: Jude Bellingham, Jordan Henderson, Mason Mount, Kalvin Phillips, Declan Rice
Forwards: Dominic Calvert-Lewin, Phil Foden, Jack Grealish, Harry Kane, Marcus Rashford, Bukayo Saka, Jadon Sancho, Raheem Sterling
* Replaces Dean Henderson, who withdrew due to injury
Goalkeepers: Georgiy Bushchan, Andriy Pyatov, Anatolii Trubin
Defenders: Eduard Sobol, Illia Zabarnyi, Serhiy Kryvtsov, Denys Popov, Oleksandr Tymchyk, Vitaliy Mykolenko, Oleksandr Karavaev, Mykola Matviyenko
Midfielders: Serhiy Sydorchuk, Ruslan Malinovskyi, Mykola Shaparenko, Marlos, Yevhen Makarenko, Oleksandr Zinchenko, Viktor Tsygankov, Taras Stepanenko, Andriy Yarmolenko, Oleksandr Zubkov, Heorhii Sudakov, Roman Bezus
Forwards: Roman Yaremchuk, Artem Dovbyk
Maguire, Foden, Rice and Phillips will be suspended for England if they pick up another yellow card in Rome, while Ukraine quartet Sergiy Sydorchuk, Mykola Shaparenko, Andriy Yarmolenko and Artem Dovbyk run the same risk.