Alex Coles, Fin Smith and Tommy Freeman started for the Red Rose, while Trevor Davison and Henry Pollock came off the bench.
But the Saints quintet were unable to stop Italy prevailing as the hosts picked up an impressive 23-18 success at Stadio Olimpico.
Freeman scored for England during the first half, but Italy were eventually able to get the job done, claiming their first ever victory against England in the Guinness Six Nations.
It meant Steve Borthwick’s side suffered a third successive defeat in this year’s tournament, after starting their campaign with a comfortable win against Wales.
England must now rebuild quickly as they face an imposing trip to Paris to take on France on Saturday night (kick-off 8.10pm).
Following the defeat in Italy, Rugby Football Union chief executive Bill Sweeney gave Borthwick a vote of confidence.
"After a 12-match winning run, these past three results have been hugely disappointing, and we feel that just as much as everyone else," Sweeney said.
"Steve and his coaching team are working tirelessly to make improvements, and we remain fully committed to supporting them and the players as they face France this weekend and then look ahead to the Nations Championship.
"Part of that support is being open about what hasn't gone right during this Six Nations and making sure everyone has a clear sense of how we move through those challenges together.
"That's something we'll be talking through and working on in the days and weeks ahead.
"We will work together to understand and rectify why we have been unable to meet the expectations and anticipation going into these games.
"England fans rightly expect a team that learns and grows through adversity, and we're confident this group will do everything they can to deliver that."
England back row forward Ben Earl also backed Borthwick.
"Just have a think about what this team's achieved since Steve took over,” Earl said. “We've gone third in the World Cup.
"We've won 12 out of 15 games, but had won 12 on the bounce. We've beaten the All Blacks for the first time in Twickenham since 2012.
"The noise is actually bonkers, I think it's crazy.
"If anyone is letting Steve down, it's us, the group.
"We need to play better at certain moments because we're playing well for 50 minutes, playing well for 40 minutes, playing well for 30 minutes.
"We've just got to put it together – and we will."
The England Under-20s had a much happier time in Italy last weekend as they won 37-17 in Treviso on Friday night.
Saints players Oliver Scola, Aiden Ainsworth-Cave and James Pater started the game, with Sonny Tonga’uiha and Jonny Weimann coming off the bench.
England can’t win the Under-20s Six Nations this year though as France are the runaway leaders in the competition.
1. Italy 23 England 18
Fin Smith showed his disappointment as England were beaten in Rome (photo by David Rogers/Getty Images) Photo: David Rogers
