Southampton were expelled from the Championship playoff final after admitting to spying on a training session of semifinal opponents Middlesbrough.
Middlesbrough have been reinstated and are set to face Hull at Wembley on Saturday for a place in the Premier League.
Southampton will also be docked four points next season after admitting to multiple breaches of regulations related to the "unauthorized filming of other clubs' training" sessions, according to a statement from the English Football League.
"An independent disciplinary commission has today expelled Southampton from the Sky Bet Championship play-offs," the EFL said.
Southampton, relegated from the Premier League last season, confirmed they would appeal the sanctions.
The EFL said the parties were working to ensure an appeal could be heard on Wednesday.
"Subject to the outcome, it could result in a further change to Saturday's fixture," the EFL said.
A member of the Southampton coaching staff was caught by Middlesbrough officials recording training on his phone.
The EFL confirmed further charges had been laid against Southampton, and that the club had also admitted observing training sessions ahead of matches against Oxford and Ipswich.
The first leg ended 0-0 before Southampton progressed with a 2-1 win after extra-time in the second leg.
Middlesbrough issued a statement welcoming the outcome of the disciplinary commission hearing.
"We believe this sends out a clear message for the future of our game regarding sporting integrity and conduct," it said.
