Two supporters from the away end at Brighton & Hove Albion's Amex Stadium were ejected and later arrested by the Sussex Police for homophobic abuse during Sunday's 2-2 draw against Wolverhampton Wanderers, the two clubs said.
The homophobic abuse came at a time when Premier League clubs were celebrating the Rainbow Laces campaign which supports LGBT people, with items ranging from captain's armbands to substitutes boards sporting rainbow colours.
Stewards removed the two supporters after they were reported for homophobic gestures and the pair were arrested by the police outside the stadium.
"It's both sad and depressing that once again we find ourselves having to address another situation of discriminatory abuse," Brighton's chief executive and deputy chairman Paul Barber said in a statement.
"Prior to kick-off, we had a huge show of support for the Rainbow Laces campaign brilliantly backed by the vast majority of people inside the stadium.
"And yet before the game is over, we are forced to deal with these two individuals, who think it's okay to behave in this way."
Sussex Police said the two were arrested "on suspicion of using threatening or abusive words or behaviour likely to cause harassment, alarm or distress" and later released.
"The two men – aged 46 and 48, and both from Bridgnorth in Shropshire – have been released under investigation pending further enquiries," it said in a statement.
Wolves strongly condemned the behaviour of the fans arrested and said they would support the Sussex Police in their criminal investigation.
Greater Manchester Police arrested a 41-year-old man over the weekend in connection with alleged racial gestures and chants in Saturday's match between Manchester City and Manchester United at the Etihad Stadium.