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- Title
Application of in vivo induced antigen technology (IVIAT) to Bacillus anthracis.
- Authors
Rollins, Sean M; Peppercorn, Amanda; Young, John S; Drysdale, Melissa; Baresch, Andrea; Bikowski, Margaret V; Ashford, David A; Quinn, Conrad P; Handfield, Martin; Hillman, Jeffrey D; Lyons, C Rick; Koehler, Theresa M; Calderwood, Stephen B; Ryan, Edward T
- Abstract
In vivo induced antigen technology (IVIAT) is an immuno-screening technique that identifies bacterial antigens expressed during infection and not during standard in vitro culturing conditions. We applied IVIAT to Bacillus anthracis and identified PagA, seven members of a N-acetylmuramoyl-L-alanine amidase autolysin family, three P60 family lipoproteins, two transporters, spore cortex lytic protein SleB, a penicillin binding protein, a putative prophage holin, respiratory nitrate reductase NarG, and three proteins of unknown function. Using quantitative real-time PCR comparing RNA isolated from in vitro cultured B. anthracis to RNA isolated from BALB/c mice infected with virulent Ames strain B. anthracis, we confirmed induced expression in vivo for a subset of B. anthracis genes identified by IVIAT, including L-alanine amidases BA3767, BA4073, and amiA (pXO2-42); the bacteriophage holin gene BA4074; and pagA (pXO1-110). The exogenous addition of two purified putative autolysins identified by IVIAT, N-acetylmuramoyl-L-alanine amidases BA0485 and BA2446, to vegetative B. anthracis cell suspensions induced a species-specific change in bacterial morphology and reduction in viable bacterial cells. Many of the proteins identified in our screen are predicted to affect peptidoglycan re-modeling, and our results support significant cell wall structural remodeling activity during B. anthracis infection. Identification of L-alanine amidases with B. anthracis specificity may suggest new potential therapeutic targets.
- Publication
PloS one, 2008, Vol 3, Issue 3, pe1824
- ISSN
1932-6203
- Publication type
Journal Article
- DOI
10.1371/journal.pone.0001824