Because of the potential bioterrorism threat, the development of a safe and effective B. pseudomallei vaccine is a national and worldwide goal. The Applied Proteomics Laboratory at UC Irvine has developed a high throughput protein expression system called PCR Express which can be used to rapidly generate complete proteomes from any sequenced microorganism, including the Class A, B & C Bioterrorism Agents. The technology allows hundreds of different genes to be expressed directly from their PCR products at the rate of hundreds of different proteins per week. Here the Burkholderia pseudomallei proteome will be generated and applied to the problem of identifying antigens recognized by B-cells and T-cells that will be useful in a subunit vaccine against the agent. The first form of the proteome will be on microarray chips which will be used to quantify serum antibody titers from BALB/c mice vaccinated with a rationally attenuated auxotroph of B. pseudomallei (2D2) and humans naturally exposed to B. pseudomallei in endemic regions of NE Thailand against each of the individual bacterial proteins. For the second proteome format, each individual protein will be purified and presented in a form that will enable them to be used in T cell restimulation assays in vitro using either spleen cells from 2D2 vaccinated mice or whole blood from exposed human populations. T cell activation will be determined by ELISA assay for secretion of lFN?. The responsive antigens identified by this Vaccine Antigen Scan will be tested for their efficacy in a murine model of infection with B. pseudomallei and, because of the extensive genetic similarity between B. pseudomallei and B. mallei, in animal models of glanders. This quantitative humoral and cellular immune response scan will produce the first complete profile of the immune response against B. pseudomallei in humans and experimental animals and will identify the most effective candidate antigens for development of a DNA or subunit vaccine against human melioidosis and glanders.
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