The movie begins at a blood bank. A homeless man walks into
the clinic and sits by another homeless man (Luke Goss) with a scar running down his chin. A nurse takes the latter man into a room with several people waiting where he is to be drained of blood, but kills his captors and reveals himself to be a new breed of vampire.
Two years have passed since the climax of the first film, and Blade (Wesley Snipes) has been trying to find Whistler (Kris Kristofferson), who survived his suicide attempt and turned into a vampire. He has been sweeping across Russia and eastern Europe, searching for his old friend and mentor, enlisting the aid of a young man, Scud (Norman Reedus), to design him a new line of gadgetry and weaponry. In the first scene, Blade fights his way through a large gang of vampires, leaving one of their number, Rush (Santiago Segura), alive, yet telling him he will be back for him. Blade finds Whistler locked in a tank of blood by a gang of vampires who were keeping him alive for purposes of torture. Blade rescues him and brings him to Prague.
Meanwhile, a crisis has arisen in the vampire community. What seems to be a more developed strain of vampirism (dubbed the "Reaper virus") is sweeping through their ranks, giving its carriers fearsome new characteristics. The original carrier of the strain is Jared Nomak (the homeless man from the beginning of the film). Far stronger than common vampires, the Reapers have three-way jaws, leech-like suckers and hearts encased in a thick layer of bone, making them invulnerable to any weapon barring sunlight. They also have a ravenous hunger, requiring more feeding than vampires, and even their dead victims become reapers. In order to combat the virus, the vampire elder/overlord Eli Damaskinos (Thomas Kretschmann) sends his minions, Asad (Danny John-Jules) and Damaskinos' daughter Nyssa (Leonor Varela), to find and strike an uneasy truce with Blade. Upon meeting Eli Damaskinos and his familiar Karl Kounen (Karel Roden), Blade learns of the vampire community's plight. They concede to him that the Reapers are the greater evil and once they finish the vampire population, they will descend on humankind.
To this end, Blade teams up with the Bloodpack, a group of vampire warriors and assassins who were originally assembled to kill him. The group consists of Rheinhardt (Ron Perlman), Chupa (Matt Schulze), Snowman (Donnie Yen), Verlaine (Marit Velle Kile), Lighthammer (Daz Crawford), and Priest (Tony Curran). The group starts at a vampire nightclub which goes disastrous when one of the Bloodpack members is bitten by Jared.
After a climactic battle against numerous Reapers in their hive within the sewers that cost the lives of some of the Bloodpack, Blade is apprehended by Damaskinos' forces, along with Whistler and Scud. It is revealed that, in his efforts to create a day-walking vampire race, Damaskinos used his own blood to create Nomak. Scud is also revealed to be one of Damaskinos' familiars. Now that the reapers are dealt with, he plans on dissecting Blade to create more daywalkers. However, Whistler escapes and rescues Blade. Blade fights his way through Damaskinos' henchmen, which ends with Rheinhardt and Scud being killed.
Meanwhile, Nomak has entered Damaskinos' stronghold seeking revenge on his "father". Just before he can escape to his helicopter, Damaskinos is betrayed by Nyssa (who became disillusioned with her father's extreme methods) and killed by Nomak when Damaskinos tries to negotiate with him. In order to "complete the circle", Nomak also bites Nyssa and then makes to leave, whereupon he is confronted by Blade. After a very brutal fight scene, Blade finds the weak spot in Nomak's physical defenses and jams his sword beneath his arm, bypassing the bone shield of his heart. Nomak then pushes it through, killing himself. With Nomak dead, Blade then carries a mortally wounded, soon-to-be-Reaper Nyssa outside for the sunrise to honor her last wish to see the sunlight, where she disintegrates in his arms.
In the film's last scene, Blade disposes of Rush, stating "Now you didn't think I'd forget about you" and stabs his sword through Rush's head.
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