- Retroactive Recognition: As a Long Runner, The F.B.I. features many actors who went on to become stars.
- Anne Archer in "Downfall".
- Edward Asner in three episodes: "The Tormentors", "The Dynasty" and "The Attorney".
- Joe Don Baker in "Summer Terror".
- Ed Begley Jr. in "The Deadly Gift".
- Karen Black in "The Satellite".
- Robert Blake in "A Mouthful of Dust" and "The Price of Death".
- Beau Bridges in "An Elephant Is Like a Rope".
- Jeff Bridges in "Boomerang".
- Charles Bronson in "The Animal" (1966 episode)
- Georg Stanford Brown in "The Intermediary".
- James Caan in "A Life in the Balance".
- David Cassidy in "The Fatal Impostor".
- Nicholas Colasanto in "The Camel's Nose" and "Bitter Harbor".
- Dabney Coleman in seven episodes: "Slow March Up a Steep Hill", "The Hijackers", "The Conspirators", "Flight", "Incident in the Desert", "The Game of Terror" and "Survival".
- Bruce Dern in "Pound of Flesh" and "The Nightmare".
- Richard Donner directed the first-season episode "The Spy Master".
- Pete Duel in "Slow March Up a Steep Hill" and "False Witness".
- Michael Douglas in "The Hitchhiker".
- Robert Duvall in six episodes: "The Giant Killer", "The Scourge", "The Executioners (Parts 1 and 2)", "The Harvest" and "Nightmare Road".
- Dana Elcar in five episodes: "The Messenger", "The Young Warriors", "The Fatal Connection", "Superstition Rock" and "Fatal Reunion".
- Sam Elliott as an FBI agent in "The Prey".
- Harrison Ford appears in two episodes, "Caesar's Wife" and "Scapegoat". In the former episode, Ford gets almost killed in a fight on the beach with, of all people, Russell Johnson!
- Meg Foster in "A Second Life" and "The Animal" (1974 episode, not to be confused with the 1966 episode of the same title).
- Mary Frann in "Confessions of a Madman".
- Leif Garrett in "The Deadly Species".
- Jonathan Goldsmith in "The Monster", "The Maze", "Flight" and "The Gopher" (all billed as Jonathan Lippe).
- Peter Graves in "Rope of Gold". It was one of his last guest roles prior to Mission: Impossible.
- Charles Grodin in "Sky On Fire".
- Gene Hackman in "The Courier".
- Mark Hamill in "The Corruptor".
- Brion James in "The Animal" (1974 episode).
- Peter Jason in "The Homecoming".
- Diane Keaton in "Death Watch".
- Harvey Keitel in "Deadly Ambition".
- Ted Knight in "An Elephant Is Like a Rope", "The Assassin" and "The Executioners (Part Two)".
- Larry Linville in "Flight" and "The Inheritors" (billed as Lawrence Linville).
- Hal Linden in "The Confession".
- Jack Lord in "Collision Course".
- Stuart Margolin in "The Homecoming".
- Gerald McRaney in "Deadly Ambition".
- Leslie Nielsen in "Pound of Flesh" and "Fools' Gold".
- Annette O'Toole in "The Lost Man".
- Robert Pine in "The Set-Up" and "Confessions of a Madman".
- Burt Reynolds in "All the Streets Are Silent" and "Act of Violence".
- Alex Rocco in "The Outcast".
- Wayne Rogers in seven episodes: "The Tormentors", "The Extortionist", "The Legend of John Rim", "The Ninth Man", "Deadfall", "The Traitor" and "Superstition Rock".
- Kurt Russell in "The Tormentors".
- John P. Ryan in "Breakthrough" and "The Swindler" (billed as John Ryan).
- Tom Selleck in "The Confession".
- Tom Skerritt in four episodes: "The Assassin", "The Legend of John Rim", "Unknown Victim" and "The Deadly Species".
- Martin Sheen in four episodes: "The Dynasty", "The Condemned", "A Second Life" and "The Disinherited".
- Bobby Sherman in "The Mechanized Accomplice".
- David Soul in "The Runner".
- Richard Thomas in "The Game of Terror".
- Daniel J. Travanti in four episodes: "Death of a Fixer", "The Diamond Millstone", "The Franklin Papers" and "Confessions of a Madman".
- Robert Urich in "The Runner".
- Joan Van Ark in four episodes: "The Maze", "The Condemned", "The Deadly Gift" and "The Vendetta".
- Lindsay Wagner in "Dark Journey".
- Billy Dee Williams in "The Eye of the Storm", "The Sanctuary" and "The Architect".
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