Two years have passed since the ending of the first film, and Blade has been keeping himself busy in the hunt for Whistler, who failed to kill himself in the original. He has been sweeping across Russia and eastern Europe searching for his old friend and mentor, enlisting the aid of a young man named Scud to design him a new line of equipment and weapons. Blade fights his way through a large gang of vampires, leaving a bloodsucker named Rush alive, yet telling him he'll be back for him. Finding Whistler locked in a tank of blood by a cruel gang of vampires who were keeping the old man 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 powers and features. The original carrier of the strain seems to be Jared Nomak, a one-time vampire who appears to have mutated into a creature even more dangerous - one that prefers the blood of vampires to human blood, in the process transforming his vampiric victims into Reapers themselves (albeit far less stable).
The Reapers bear a certain resemblance to Blade, being far stronger than common vampires and lacking an allergy to silver, but they also possess radical physical mutations: three-way jaws, leech-like tongues and hearts encased in a thick layer of bone at the front and back – in short, their only vulnerability is ultraviolet light. However, all these advantages are countered by an incredibly fast metabolism - they must feed almost constantly to avoid starving to death. It is why they prefer vampire blood to human blood - it possesses even more nutrition. And for all Reapers save Nomak, it is a losing battle - they still sicken and die shortly after being turned.
In order to combat the virus, the vampire elder / overlord Eli Damaskinos and his familiar servant Carter Counan (a lawyer) send their minions Asad and Nyssa (who is Damaskinos' daughter) to find and strike an uneasy treaty with Blade, convincing him with the argument that though vampires kill humans for food, they do so only in a controlled fashion. Reapers are even more dangerous as they feed almost constantly and once they finish the vampire population, they will doubtlessly descend on humankind. Whilst Blade may hate vampires, the Reapers are far more dangerous and neither side can attack them without uniting first.
To this end, Blade teams up with the Bloodpack, a group of vampire warriors and assassins who were originally assembled to kill Blade. The vampires are Reinhardt, Nyssa, Asad, Priest, Chupa, Lighthammer, Snowman and Verlaine. In order to obtain some measure of control over the group, Blade singles out Reinhardt (played by Ron Perlman), one of the group's more resistant members, and installs a remotely-activated explosive device in the back of his head. Setting their mutual hatred aside, Blade leads the Bloodpack in the fight against the Reapers and the investigation into their origins. During this time, Blade forms something of an intimate relationship with Nyssa, the daughter of Damaskinos and member of the Bloodpack. In the nightclub, Blade and the Bloodpack battle the Reapers, the Bloodpack vampire Priest being bitten and killed as he changes into a Reaper. Lighthammer is also bitten but does not turn.
In the sewers, Blade splits the team into groups. Reinhardt and Chupa corner Whistler, and Chupa proceeds to savagely beat Whistler for the death of Priest: "We lose a partner, and Blade loses one!" Reinhardt abandons Chupa, hunting for more Reapers. Whistler releases a batch of Reaper pheromones into the air in a bid to save himself. Just as Whistler is nearly finished, the Reapers attack and maul Chupa to death, allowing Whistler to escape. Meanwhile, Lighthammer transforms into a Reaper and feeds on, and kills, Snowman. His lover Verlaine gets him to chase her up a manhole, so that when she removes the cover, both of them are killed by the sunlight. When the Reapers kill Asad, it's left to Reinhardt, Nyssa, Blade and Whistler to escape.
After a climactic battle, Blade activates a UV Bomb that incinerates all the Reapers except Nomak who passes information along to Whistler. Blade is apprehended by Damaskinos' forces, along with Whistler and Scud. As it turns out, the Reaper strain is not a virus at all, but rather a genetic experiment gone wrong. In his efforts to replicate Blade – a day-walking silver-immune vampire, Damaskinos had numerous experiments performed on Nomak, who is in fact his estranged son. He remarks that Nomak was the first carrier of the strain but ultimately flawed, seeing as he was vulnerable to daylight, as were all the other carriers whom he'd infected. Damaskinos then reveals another horrible truth — he has been creating many more vampire prototypes, intended as the next step of vampire evolution, all of them kept in fetal form in a massive incubator. All they require now is Blade's biological make-up to give them protection from the sun, and the vampire nation will all be Daywalkers. In order to extract this, Damaskinos plans to have Blade killed and dissected.
During his captivity, Blade attempts to activate the pre-placed explosive in Reinhardt's skull, but Scud reveals that the bomb (which he himself had crafted) was never designed to go off. He himself is in fact one of Damaskinos' familiars and planned to side with the vampires rather than fighting against them. Figuring that it would all be over for the human race once all vampires were immune to daylight, he decided "[he] would rather be a pet than cattle." However, Blade has a trump card - he has always suspected Scud's servitude to Damaskinos, and his backhanded double dealing. Activating a second switch on his remote, Blade kills the unfortunate Scud, who unluckily happened to be holding the bomb at the time. After Whistler rescues him from the dissection (after escaping from Reinhardt with a gun himself) by killing the familiar watching over Blade, Blade fights his way through Damaskinos' henchmen, and after bisecting Reinhardt with his sword, heads towards the lead vampire himself while Whistler destroys the vampire fetii by shooting them.
Meanwhile, a vengeful Nomak has entered Damaskinos' stronghold, seeking revenge on the father who mutilated him and turned him into the first Reaper. Just before he can escape, Damaskinos is betrayed by Nyssa (who became disillusioned with her father's extreme methods) and killed by Nomak. In order to "complete the circle", Nomak also bites Nyssa and then makes to leave, whereupon he is confronted by Blade. After a very physical fight scene, Blade finds the weak spot in Nomak's physical defenses and jams his sword beneath his arm, penetrating his heart from the side, bypassing the bone shield on his heart. Blade then falls to the floor, nearly finished due to broken ribs and internal bleedings sustained during the fight. Severely injured, Nomak crawls away and props himself up against a pillar. He is mortally wounded, so he puts himself out of his misery by shoving the broken sword the final inch into his heart. Before he dies he comments that “it hurts no more” (during the film the Reaper Virus seems to cause him great pain).
With Nomak dead, Blade then carries Nyssa outside for the sunrise at her request, where she disintegrates in his arms before she becomes a Reaper.
In the film's final scene, the vampire Rush is in a London strip club, preparing to make use of one of the booths. However, as soon as the curtains open, he is shocked to find Blade on the other side of the glass, who casually remarks "You didn't think I'd forget about you, did you?", and stabs his sword into his skull.
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