BACKGROUND Transcutaneous immunization (TCI) is a needle-free technique that delivers antigens and adjuvants to potent epidermal immune cells. To address critical unmet needs in biodefense against anthrax, we have designed a novel vaccine delivery system using a dry adhesive patch that simplifies administration and improves tolerability of a subunit anthrax vaccine. METHODS Mice and rabbits were vaccinated with recombinant protective antigen of Bacillus anthracis and the heat-labile toxin of Escherichia coli. Serologic changes, levels of toxin-neutralizing antibodies (TNAs), and pulmonary and nodal responses were monitored in the mice. A lethal aerosolized B. anthracis challenge model was used in A/J mice, to demonstrate efficacy. RESULTS The level of systemic immunity and protection induced by TCI was comparable to that induced by intramuscular vaccination, and peak immunity could be achieved with only 2 doses. The addition of adjuvant in the patch induced superior TNA levels, compared with injected vaccination. CONCLUSIONS Anthrax vaccine patches stimulated robust and functional immune responses that protected against lethal challenge. Demonstration of responses in the lung suggests that a mechanism exists for protection against challenge with aerosolized anthrax spores. A formulated, pressure-sensitive, dry adhesive patch, which is stable and can be manufactured in large scale, elicited comparable immunoglobulin G and TNA responses, suggesting that an anthrax vaccine patch is feasible and should advance into clinical evaluation.

Human involucrin (hINV) is assembled into cornified structures via formation of transglutaminase (TG)-dependent interprotein ε-(γ-glutamyl)lysine bonds. The hINV sequence includes 150 glutamine residues that could function as potential sites of cross-link formation. The present studies were designed to evaluate the extent to which hINV can function as a TG substrate under optimal conditions and in the absence of other substrates. Incubation of hINV with TG results in formation of 4–5 isopeptide bonds per hINV molecule. When the small amine donor 14C-putrescine is included in the reaction, 48 Q residues are labled. Isotope distribution and sequence analysis suggests that the 14C-putrescine-labeled sites are located throughout the protein. Our present results show that many hINV Q residues can be utilized for cross-link formation, and that hINV can be cross-linked at very high cross-link densities. These results suggest that, in vivo, factors other than hINV structure limit the number of residues used for cross-link formation.