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Molecular & Cellular Proteomics 8:924-935, 2009.
© 2009 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

Extensive Antibody Cross-reactivity among Infectious Gram-negative Bacteria Revealed by Proteome Microarray Analysis ^*

Sarah L. Keasey, Kara E. Schmid, Michael S. Lee, James Meegan, Patricio Tomas^¶, Michael Minto^¶, Alexander P. Tikhonov^¶, Barry Schweitzer^¶ and Robert G. Ulrich^,||

From the Army Medical Research Institute of Infectious Diseases, Frederick, Maryland 21702, Invitrogen Federal Systems, Frederick, Maryland 21704, and ^¶ Invitrogen Corporation, Branford, Connecticut 06405

Antibodies provide a sensitive indicator of proteins displayed by bacteria during sepsis. Because signals produced by infection are naturally amplified during the antibody response, host immunity can be used to identify biomarkers for proteins that are present at levels currently below detectable limits. We developed a microarray comprising 70% of the 4066 proteins contained within the Yersinia pestis proteome to identify antibody biomarkers distinguishing plague from infections caused by other bacterial pathogens that may initially present similar clinical symptoms. We first examined rabbit antibodies produced against proteomes extracted from Y. pestis, Burkholderia mallei, Burkholderia cepecia, Burkholderia pseudomallei, Pseudomonas aeruginosa, Salmonella typhimurium, Shigella flexneri, and Escherichia coli, all pathogenic Gram-negative bacteria. These antibodies enabled detection of shared cross-reactive proteins, fingerprint proteins common for two or more bacteria, and signature proteins specific to each pathogen. Recognition by rabbit and non-human primate antibodies involved less than 100 of the thousands of proteins present within the Y. pestis proteome. Further antigen binding patterns were revealed that could distinguish plague from anthrax, caused by the Gram-positive bacterium Bacillus anthracis, using sera from acutely infected or convalescent primates. Thus, our results demonstrate potential biomarkers that are either specific to one strain or common to several species of pathogenic bacteria.

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