Ipswich Town say anyone convicted of homophobic chanting "will not be welcomed back" at the club.
The statement comes as the British Transport Police are looking to speak to five men after an alleged homophobic incident on a train between Plymouth and Paddington on Sunday, October 31.
The previous day, Ipswich Town had lost 2-1 in a League One fixture at Plymouth Argyle's Home Park.
Club bosses said they have a "zero-tolerance" approach to homophobic behaviour and confirmed the club is working with police regarding the alleged incident.
A spokesman for Ipswich Town said: "The club has been aware and has been dealing with this incident together with British Transport Police and Suffolk police over the last six weeks.
"At Ipswich Town Football Club we believe everyone has the right to enjoy attending matches and following their club in a safe, tolerant and friendly environment.
"To safeguard the reputation the club has built up over the years, and to ensure our aims are met, we will continue to adopt a zero-tolerance approach to any such homophobic, racist or anti-social behaviour."
The club said anyone found guilty of such offences "will not be welcomed back at our club".
Police believe the men in the images may be able to help them with their investigation.
If you recognise them, or have any information, please contact the British Transport Police by texting 61016 or calling 0800 40 50 40 quoting reference 2100082301.
Alternatively, you can call Crimestoppers anonymously on 0800 555 111.
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