Kristan Ann Ware, 27, who quit the Miami Dolphins' after three years, said she felt forced to after a 2016 interview with the team director where her sexual history was discussed.
Speaking to the BBC, she explained why she submitted her complaint to the Florida Commission on Human Relations.
"I am not against them. I just want to make it a better place for the cheerleaders," she said.
She said the bullying began after an incident on a coach in London took place in 2015.
Ms Ware claimed after her team played the New York Jets at Wembley Stadium, the dancers talked about their "sex playlist".
Her complaint states she ""was pushed to give her playlist," which prompted her to admit she was a virgin.
Her lawyer Sara Blackwell said: "Kristan told her teammates that she was waiting until she was married because of her personal relationship with God."
Ms Ware alleges the team director, Dorie Grogan, confronted her about the claim during a yearly performance review and was told not to discuss her virginity at all in the future.
As part of her claim, Ms Ware also argues the team's Instagram account did not promote any of her posts that mentioned her Christian faith and a story she wrote for the Dolphins' blog had all her references to God and Christ edited out.
The Miami Dolphins said in a statement to the BBC: "We are seriously committed to providing a positive work environment for everyone associated with the organisation.
"We hold every member of our organisation to the same standards and do not discriminate as it relates to gender, race and religious beliefs."
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