The Larry Sanders Show included LGBTQ+ characters in a largely incidental manner. While not overtly negative, the narrative often focused on the reactions and discomfort of straight characters rather than deeply exploring or affirming queer identities. The portrayals were present but not central, resulting in a neutral overall impact.

The show features a transsexual character, Darlene, who dates Hank Kingsley. Upon discovering her identity, Hank reacts with shock and discomfort, leading to their breakup. The portrayal is negative, as the show uses Hank's prejudiced reaction as a source of comedy without offering a clear critique or counter-narrative, thereby normalizing transphobia.

The show frequently depicts characters who profess Christian faith as hypocritical, superficial, or using religion for public image rather than genuine belief. The narrative consistently exposes these performative aspects without offering significant counterbalancing portrayals of sincere or positive Christian adherence.

Larry Sanders' Jewish heritage is portrayed as an authentic and integral part of his character's identity, often explored through self-deprecating humor and cultural references. The show treats this aspect with nuance and personal connection, without satirizing the religion itself.