Eric Dier will miss England's Nations League match against Denmark on Wednesday through injury.
The Tottenham defender sustained a slight hamstring complaint during the 2-1 victory against Belgium on Sunday.
Dier, 26, has not been included in Gareth Southgate's 23-man squad for the visit of Denmark at Wembley Stadium as England aim to extend their unbeaten campaign in Group A2.
Harry Winks, Michael Keane, Harvey Barnes and Tammy Abraham have also been left off the squad list published by UEFA.
Abraham was a late arrival in linking up with the England squad after footage emerged of him at a party flouting the government's 'rule of six' regulations along with fellow England players Ben Chilwell and Jadon Sancho.
Injury led to Chilwell's withdrawal from the squad on Tuesday, when Kieran Trippier also left the camp to deal with a Football Association hearing for alleged betting breaches.
Southgate said on Tuesday that England "do not risk players" after Sky Sports News reported about a potential club versus country row brewing between Tottenham and the national side over the availability of Harry Kane, who is nursing muscle fatigue.
Spurs boss Jose Mourinho expressed concerns over Kane's workload before this month's international break but Southgate insisted the striker has not suffered from a specific injury and is line to feature against Denmark.
"He was feeling delayed muscle soreness, which can often happen to athletes but there was no injury at any point," Southgate said.
"Our issue with Harry before Sunday's game was to be able to perform at the highest level. Now, with the addition of a couple of days' training, it's not even an issue.
"He's been fully fit since the end of last week, just before the last game. He would have started the last game but hadn't had enough training time.
"He was obviously fine during the game so he's fit and available."
Southgate's England in tournament mode
Southgate's back three is not bringing many thrills and spills but it signals a return to the sort of risk-free football that wins international tournaments, writes Sky Sports' Adam Bate.
Watch England vs Denmark live on Sky Sports Main Event from 7pm on Wednesday; kick-off 7.45pm.