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Biology subjects

Scott, T. A.

Publications and source records attributed to Scott, T. A..

2 recordsLinked to original sources

Protective and pathogenic antibody responses from a primate Shigella outbreak inform vaccine design

There is currently no approved vaccine for Shigella spp., a leading cause of diarrhea that are increasingly resistant to antimicrobials. Shigella vaccine development is complicated in part by an incomplete understanding of the structural and molecular determinants of immunity. To address this, we isolated monoclonal antibodies (mAbs) against candidate Shigella vaccine antigens using samples from a Shigella flexneri outbreak in a non-human primate (NHP) research facility. We found that antibodies targeting the Shigella O-antigen (O-Ag) can undergo significant affinity maturation (>10%) to acquire broad cross-reactivity across S. flexneri serotypes. We also found that the virulence-associated type III secretion system (T3SS) proteins IpaD and IpaB elicit moderate T cell and robust antibody responses. T3SS antibodies either inhibit, or - surprisingly - enhance bacterial virulence in vitro and in vivo depending on their epitope specificity. These findings provide key insights into protective and deleterious immune responses against Shigella that directly inform vaccine immunogen design.

immunology↗

Self-immunity guided identification of threonyl-tRNA synthetase as the molecular target of obafluorin, a β-lactone antibiotic

To meet the ever-growing demand of antibiotic discovery, new chemical matter and antibiotic targets are urgently needed. Many potent natural product antibiotics which were previously discarded can also provide lead molecules and drug targets. One such example is the structurally unique {beta}-lactone obafluorin, produced by Pseudomonas fluorescens ATCC 39502. Obafluorin is active against both Grampositive and -negative pathogens, however the biological target was unknown. We now report that obafluorin targets threonyl-tRNA-synthetase and we identify a homologue, ObaO, which confers self-immunity to the obafluorin producer. Disruption of obaO in P. fluorescens ATCC 39502 results in obafluorin sensitivity, whereas expression in sensitive E. coli strains confers resistance. Enzyme assays demonstrate that E. coli threonyl-tRNA synthetase is fully inhibited by obafluorin, whereas ObaO is only partly susceptible, exhibiting a very unusual partial inhibition mechanism. Altogether, our data highlight the utility of a self-immunity guided approach for the identification of an antibiotic target de novo and will ultimately enable the generation of improved obafluorin variants.

microbiology↗