Indiana Jones and the Last Crusade does not feature any identifiable LGBTQ+ characters or themes. The film's narrative focuses on heterosexual relationships and the quest for the Holy Grail, with no elements related to queer identity present in its plot or character arcs.

Indiana Jones and the Last Crusade does not feature any identifiable transsexual characters or themes. The narrative is entirely focused on an adventure quest for the Holy Grail, the conflict with Nazis, and the complex relationship between Indiana Jones and his father, Henry Jones Sr.

The film features Dr. Elsa Schneider as a prominent female character, but her role involves manipulation and intellectual pursuits rather than direct physical combat. No female character is depicted defeating male opponents in close-quarters physical altercations.

All established characters maintain their original genders, and new characters introduced in the film do not represent gender swaps of pre-existing figures. There are no instances of a character canonically or historically established as one gender being portrayed as another.

All major characters in "Indiana Jones and the Last Crusade" are either new and portrayed consistently with their apparent race, or are returning characters portrayed by the same actors or actors of the same race as previously established. No characters were canonically, historically, or widely established as one race and then portrayed on screen as a different race.