Hull City's crowd pilot on Saturday has been cancelled amid a rising number of Covid-19 cases in the city.
The League One game against Crewe was one of eight in the English Football League allowing supporters to attend.
But with infection rates in Hull more than tripling in the past week, the local council has cancelled the government-led pilot.
Up to 1,000 fans will still be allowed at the seven other games and they will be the first EFL crowds in six months.
All games in the Championship, League One and League Two have been behind closed doors since March, although 872 fans attended an EFL Trophy match at Cambridge United earlier this month.