Raheem Sterling, Leighton Baines and James Milner will not play in Tuesday's friendly in Italy with Danny Welbeck also a concern after picking up knocks in the win against Lithuania.
It was a largely jubilant evening for England with Harry Kane scoring on his debut in the 4-0 win in their European Qualifier at Wembley.
However, it wasn't all good news with Liverpool forward Sterling suffering a toe injury, Manchester City's Milner carrying a knee problem and left-back Baines set to return to his club Everton.
England boss Roy Hodgson was delighted with his team's showing but expressed particular concern over two of his star performers on the night at Wembley - Arsenal striker Welbeck, who scored just before half-time, and Sterling, stretchered off late on.
"It all went right apart from Welbeck's injury towards the end of the game which has soured it a bit," Hodgson said.
"We hope it's not serious, but it'll probably keep him out of Tuesday, which is a disappointment particularly as we'd already lost Daniel Sturridge (through injury on Monday).
"Welbeck has had a tweak to the knee, when he dived with the goalkeeper. We were planning to take him off anyway. He's being assessed by the medical staff, but we won't get a proper idea until Saturday.
"The medical staff gave me the impression they're not terribly worried about it, but that Tuesday may be too soon. But we'll see."
"It is two blows for us," Hodgson told ITV. "Raheem Sterling was waiting for an injection on his toe - he has been waiting for a while.
"He's been playing through a situation, getting himself ready each week to get on the field of play. He did it again tonight, which is terrific. But he doesn't need to continue those shows of bravery. It's important he gets it put right for us and Liverpool.
"They think this injection will help him get through the rest of the season and join us and beat Slovenia (in June).
"If we're unlucky we will lose him and Danny Welbeck (for the game against Italy). They're important players in the system we're trying to play, but it will give opportunities for other players."
Hodgson confirmed that uncapped Southampton left-back Ryan Bertrand has been invited to join the squad for Tuesday's friendly in Turin. Hodgson will have 21 players for the match in the country where he enjoyed spells with Inter Milan and Udinese.
"We'll be short of a few players for a game that means a lot to us and to me," he added. "But I'm delighted with tonight's performance and for Harry Kane."