The Sacrifice takes place between the Blood Harvest campaign of the main Left 4 Dead game, and leads up to the events of The Sacrifice downloadable campaign for Left 4 Dead. Interspersed within the story are flashbacks showing the events that each survivor witnessed at the onset of the infection, watches friends, loved ones, and strangers turn into zombie-like creatures before their eyes and each realizing they are immune to the infection.

From the end of Blood Harvest, the four survivors, Bill, Zoey, Louis, and Francis, are rescued by U.S. military and taken to a well-fortified safe zone. They are immediately split up and put into isolation rooms when they are learned to be carries of the infection. They are tested and debriefed by the military group. They describe to Lt. Mora how the infection has mutated to create special infected with unique abilities; though Lt. Mora agrees with the survivors reports, his superior, Everly, is highly skeptical, and refuses to take steps to consider evacuating the base despite not receiving orders for the last nine days. Mora becomes concerned that the survivors are being rounded up to find a cure for the infection, and will be killed if none is found, and privately discussions plans to help the survivors to escape.

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