Tyler Adams and Giovanni Reyna scored as the United States beat Mexico to win the Concacaf Nations League.
The game in Arlington, Texas, was paused three times because of homophobic chanting from fans.
Concacaf, the governing body for football in North and Central America and the Carribbean, condemned the chanting and said a “significant number” of supporters were ejected.
The US, Mexico and Canada are joint hosts for the 2026 World Cup.
Play was stopped in the 86th minute and twice in injury time because of discriminatory chanting from the crowd when USA keeper Matt Turner took his goal-kicks.
Homophobic chanting also caused the last meeting between the sides to be paused in June 2023.
“It is extremely disappointing that this matter continues to be an issue at some matches, particularly in the context of the next two years presenting such a tremendous opportunity to grow the sport in our region,” Concacaf said.
“We will continue to urge fans to support their teams positively and with respect for the opposition and all participants in the game.”
The US have won all three Concacaf Nations League titles since its inception in 2019-20.
Midfielder Adams has spent the majority of the season on the sidelines since signing for Bournemouth in the summer after two surgeries on his hamstring.
The 25-year-old opened the scoring in just his fourth appearance this campaign but was withdrawn at half-time.
Reyna, who joined Nottingham Forest on loan from Borussia Dortmund in January, wrapped up the win with his eighth goal for the national side.
“It never gets old winning titles for your country. I’ll never take any of them for granted,” midfielder Reyna said.
“The group of guys and the energy we have, I always love coming to camp. I’m at a loss for words right now.”
