Treatment of experimental anthrax with pegylated circularly permuted capsule depolymerase.

Abstract

Anthrax is considered one of the most dangerous bioweapon agents, and concern about multidrug-resistant strains has led to the development of alternative therapeutic approaches that target the antiphagocytic capsule, an essential virulence determinant of <i>Bacillus anthracis</i>, the causative agent. Capsule depolymerase is a γ-glutamyltransferase that anchors the capsule to the cell wall of <i>B. anthracis</i>. Encapsulated strains of <i>B. anthracis</i> can be treated with recombinant capsule depolymerase to enzymatically remove the capsule and promote phagocytosis and killing by human neutrophils. Here, we show that pegylation improved the pharmacokinetic and therapeutic properties of a previously described variant of capsule depolymerase, CapD-CP, when delivered 24 hours after exposure every 8 hours for 2 days for the treatment of mice infected with <i>B. anthracis.</i> Mice infected with 382 LD₅₀ of <i>B. anthracis</i> spores from a nontoxigenic encapsulated strain were completely protected (10 of 10) after treatment with the pegylated PEG-CapD-CP^S334C, whereas 10% of control mice (1 of 10) survived with control treatment using bovine serum albumin (<i>P</i> < 0.0001, log-rank analysis). Treatment of mice infected with five LD₅₀ of a fully virulent toxigenic, encapsulated <i>B. anthracis</i> strain with PEG-CapD-CP^S334C protected 80% (8 of 10) of the animals, whereas 20% of controls (2 of 10) survived (<i>P</i> = 0.0125, log-rank analysis). This strategy renders <i>B. anthracis</i> susceptible to innate immune responses and does not rely on antibiotics. These findings suggest that enzyme-catalyzed removal of the capsule may be a potential therapeutic strategy for the treatment of multidrug- or vaccine-resistant anthrax and other bacterial infections.