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Louis, MO 63110 Timing: Fiscal Year 2002; Project Start 01-SEP-2002; Project End 31-AUG-2004 Summary: (provided by applicant): The zenith of the disease smallpox and its eradication in 1977 from human populations occurred prior to the modern era of immunology and molecular biology. Consequently there is little knowledge concerning the immune correlates for recovery from smallpox or the cross-reactive proteins expressed by baccinia virus that were responsible for its success as the smallpox vaccine.

Ebola). Accurate and efficient techniques to identify and diagnose these agents are severely limited. S. to counter future bioterrorism attacks. Available older techniques have proven unreliable. Modern molecular tests like individual PCR assays have been developed for some agents. These offer increased speed and sensitivity but because there are so many bioterrorism agents it is prohibitive to run dozens of "singleplex" arrays on each specimen. Similarly, recently reported microchip (MAGI Chip) arrays and other microarrays suffer from either needing PCR amplification first, or from the high cost to make the arrays, and the need for sophisticated equipment.

Cause for further alarm is that poxviruses, among a variety of pathogens, may relatively easily be engineered to increase their virulence, often rendering host immune responses ineffective. Such 'stealth' viruses may even overcome immunity in previously vaccinated individuals. The primary focus of this application is to evaluate novel strategies against highly virulent poxviruses and against stealth viruses with a capacity to suppress host antiviral immunity. Recently we developed a consecutive DNA/poxvirus "prime-boost" protocol that induces unprecedented levels of antiviral immunity of high avidity for the immunizing antigen.

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