The film features several LGBTQ+ characters, including the established gay couple Tweek and Craig, who are portrayed as stable and successful adults. Stan Marsh is also shown in a relationship with a transgender woman, Wendy, which is treated as a normal aspect of his life. The depictions are generally affirming, presenting these characters and relationships with dignity and without negative commentary or stereotypes.

The film features Nicole, a trans man, as a minor character married to Wendy Testaburger. His trans identity is present but not central to the narrative, avoiding both problematic stereotypes and explicit affirmation. The portrayal is incidental, neither uplifting nor denigrating.

The film portrays Eric Cartman's genuine conversion to Judaism and his loving, devout Jewish family life in a positive light, challenging audience expectations and Kyle's cynicism.