The 31-year-old Swede challenged referee Craig Pawson's decision to dismiss him for a 43rd-minute challenge on Ander Herrera during Sunday's 3-0 defeat at the Stadium of Light, but Football Association disciplinary chiefs have backed the official.
An FA statement said: "An Independent Regulatory Commission has rejected a wrongful dismissal claim from Sunderland's Sebastian Larsson.
"The midfielder was dismissed for serious foul play during the Black Cats' Premier League fixture against Manchester United on 9 April, 2017. Larsson's three-match suspension remains in place."
The news will come as a blow to manager David Moyes, who insisted after the game that Larsson's tackle did not even warrant a yellow card.
Moyes said: "I don't want to make you think it's the referee's fault, it's not that. But he gets the ball, he touches the ball. There's no contact at all, so it's not even a booking."
Larsson will miss the home games against West Ham and Bournemouth either side of a derby trip to Middlesbrough before returning for the final four fixtures of the campaign.