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Professor Alexander Saxton (Christopher Lee) is an anthropologist returning home to Europe via the Trans-Siberian Railway bringing with him a 'fossil' in a crate. The 'fossil' is the frozen remains of a primitive humanoid creature, which Saxton is sure is the missing link in the evolutionary chain. Also on board is Dr. Wells (Peter Cushing), another Englishman and a colleague of Saxton's.

Mysterious deaths occur even before the train sets out. A known thief is found dead on the platform after having looked into the crate, bleeding from his eyes, which have turned blank and white. A slightly unbalanced monk (a Rasputin-esque figure, played by the Argentine actor Alberto de Mendoza), who acts as a spiritual advisor to the Count and Countess waiting to board the train, claims that the crate is evil.

Saxton is eager to keep his find a secret from everyone, especially Dr. Wells, but Wells pays the baggage man to look into the crate. When he does, he is also killed by the glowing red gaze of the creature, which escapes the confines of its crate. The murders continue while the humanoid fossil haunts the moving train, the victims found with blank white eyes, and an autopsy leads Wells and his assistant to hypothesize that the brains of the victims are being drained of memories and knowledge. When the fossil is gunned down by Inspector Mirov, an authority in charge of the train, the threat seems to have been vanquished. However, Saxton and Wells discover images embedded in liquid inside the eye of the dead fossil, images that reveal a prehistoric Earth, as well as a view of the Earth from outer space. They deduce that the real threat is a formless alien creature that was simply inhabiting the body of the fossil, and that it has now transferred itself to the Inspector. The monk senses the evil presence inside of the Inspector and pledges allegiance to it, considering it Satan.

When news of the murders on the train is wired to Siberian authorities, the train is stopped and an intimidating cossack named Captain Kazan (Telly Savalas) boards with a small group of his men. Mirov is discovered to be the creature and is shot down, but not before the alien presence transfers itself to the monk. The passengers all retreat to the caboose, while the monk murders Kazan and his men by draining them of their brains.

Saxton rescues the Countess from the creature, but it resurrects all of its victims as white-eyed zombies. The zombies chase Saxton and the Countess back to the caboose, where the others are waiting. As they desperately work to detach the caboose from the rest of the train, Russia sends a telegraph to the next station ahead that they are to send the train over a cliff. The operators follow this order, thinking that they may be at war. Just as the surviving passengers manage to separate themselves from the rest of the train, it goes crashing over the cliff into a fiery explosion below. The caboose rolls to a safe stop precariously near the edge, where the survivors watch the fire consume the train and the unnatural inhabitants within it.

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