Abstract
Blood coagulation often accompanies bacterial infections and sepsis and is generally accepted as a consequence of immune responses. Though many bacterial species can directly activate individual coagulation factors, they have not been shown to directly initiate the coagulation cascade that precedes clot formation. Here we demonstrated, using microfluidics and surface patterning, that the spatial localization of bacteria substantially affects coagulation of human and mouse blood and plasma. Bacillus cereus and Bacillus anthracis, the anthrax-causing pathogen, directly initiated coagulation of blood in minutes when bacterial cells were clustered. Coagulation of human blood by B. anthracis required secreted zinc metalloprotease InhA1, which activated prothrombin and factor X directly (not via factor XII or tissue factor pathways). We refer to this mechanism as 'quorum acting' to distinguish it from quorum sensing—it does not require a change in gene expression, it can be rapid and it can be independent of bacterium-to-bacterium communication.
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Acknowledgements
This work was supported in part by the US National Institutes of Health (NIH) Director's Pioneer Award (grant number DP1OD003584), the US National Science Foundation CAREER Award (grant number CHE-0349034), the US Office of Naval Research (grant number N000140610630), the Camille Dreyfus Teacher-Scholar Awards Program and the Cottrell Scholar of Research Corporation Awards Program to R.F.I., by NIH grants (GM 62548 and GM 81539) to W.-J.T., and by the Intramural Research Program of the NIAID. We thank J. Alverdy, B. Bishop, S. Crosson, C. Esmon, M. Mock, M. Runyon, J. Shapiro, U. Spitz, T. Van Ha, D. Wiebel and O. Zaborina for helpful discussions; O. Zaborina (The University of Chicago) for the gift of the EGPF plasmid for E. coli and for assisting in the transformation procedure; M. Mock (Institut Pasteur) for the gift of the mouse anti-InhA1 serum; L. Cheng and D. Crown for assisting in the animal studies; H. Herwald (Lund University) for the gift of the E. coli Ymel-1 strain; J. Handelsman (University of Wisconsin) for the gift of the B. cereus GFP strain; M. Blaser (New York University) for the gift of the B. anthracis ΔluxS strain; and J. Price for contributions in writing and editing this manuscript. We thank C. Tallant (Institut de Biologia Molecular de Barcelona) for assistance in construction of the protease gene knockout strains.
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C.J.K., J.Q.B., M.M., Y.B., R.R.P., T.R.K. and F.S. performed experiments; C.J.K., J.Q.B., M.M., Y.B., R.R.P., T.R.K., F.S., S.H.L., W.-J.T. and R.F.I. designed experiments and analyzed data; C.J.K., W.-J.T. and R.F.I. wrote the paper; A.P.P. and P.S. provided reagents.
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Kastrup, C., Boedicker, J., Pomerantsev, A. et al. Spatial localization of bacteria controls coagulation of human blood by 'quorum acting'. Nat Chem Biol 4, 742–750 (2008). https://doi.org/10.1038/nchembio.124
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DOI: https://doi.org/10.1038/nchembio.124
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