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* Animal Models and Retroviral Vaccines Section, National Cancer Institute, Bethesda, MD 20892;
Southern Research Institute, Frederick, MD 21701;
Advanced BioScience Laboratories, Kensington, MD 20895;
Vaccine Research Center, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892;
¶ Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892;
|| Department of General and Experimental Pathology, Medical University of Bialystok, Bialystok, Poland;
# Biostatistics and Data Management Section, National Cancer Institute, Bethesda, MD 20892;
** U.S. Army Medical Research Institute of Infectious Diseases, Fort Detrick, MD 21702; and
Biodefense Clinical Research Branch, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892
The smallpox vaccine Dryvax, a live vaccinia virus (VACV), protects against smallpox and monkeypox, but is contraindicated in immunocompromised individuals. Because Abs to VACV mediate protection, a live virus vaccine could be substituted by a safe subunit protein-based vaccine able to induce a protective Ab response. We immunized rhesus macaques with plasmid DNA encoding the monkeypox orthologs of the VACV L1R, A27L, A33R, and B5R proteins by the intradermal and i.m. routes, either alone or in combination with the equivalent recombinant proteins produced in Escherichia coli. Animals that received only DNA failed to produce high titer Abs, developed innumerable skin lesions after challenge, and died in a manner similar to placebo controls. By contrast, the animals vaccinated with proteins developed moderate to severe disease (20–155 skin lesions) but survived. Importantly, those immunized with DNA and boosted with proteins had mild disease with 15 or fewer lesions that resolved within days. DNA/protein immunization elicited Th responses and binding Ab titers to all four proteins that correlated negatively with the total lesion number. The sera of the immunized macaques recognized a limited number of linear B cell epitopes that are highly conserved among orthopoxviruses. Their identification may guide future efforts to develop simpler, safer, and more effective vaccines for monkeypox and smallpox.
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