Manchester United striker Marcus Rashford returns to the first-team after missing England's 1-0 win over Slovakia last month.
And the 18-year-old has been joined by United team-mate Jesse Lingard, who could make his international debut after an impressive start to the season.
In a surprise move, Southgate has also named Stoke's Glen Johnson as part of the squad, with the defender called up for the first time since the 2014 World Cup.
He replaces Liverpool's Nathaniel Clyne, who is sidelined through injury, while Arsenal's Alex Oxlade-Chamberlain replaces Adam Lallana.
Speaking to TheFA.com, Southgate revealed his thinking behind his first squad.
"We've tried to keep some continuity from last month because my view would be that there are a lot of things that have been going right," he said.
"We've had a few people unavailable through injury like Harry Kane and Jack Butland, and we've lost a couple over the weekend in Danny Drinkwater, Adam Lallana and Nathaniel Clyne while Luke Shaw isn't quite back fit yet. That's had a bearing on the squad.
"There are obviously a couple of players that I've worked with in the Under-21s with Marcus Rashford and Jesse Lingard, who we think are exciting young players who we wanted to put in this squad.
"It's obviously been a difficult week in a number of ways, however we are in a position now where it's important to get the squad named. I'm looking forward now to working with the players because it's important that we build on the performance and the result against Slovakia."
West Ham's Michail Antonio, picked last month by old boss Sam Allardyce, retains his place in the side.
Raheem Sterling and Theo Walcott have been rewarded for their bright starts to the season while captain Wayne Rooney makes the cut despite being dropped by United boss Jose Mourinho.