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Maltodextrin-based imaging probes detect bacteria in vivo with high sensitivity and specificity

Abstract

The diagnosis of bacterial infections remains a major challenge in medicine. Although numerous contrast agents have been developed to image bacteria, their clinical impact has been minimal because they are unable to detect small numbers of bacteria in vivo, and cannot distinguish infections from other pathologies such as cancer and inflammation1,2,3,4,5,6,7. Here, we present a family of contrast agents, termed maltodextrin-based imaging probes (MDPs), which can detect bacteria in vivo with a sensitivity two orders of magnitude higher than previously reported, and can detect bacteria using a bacteria-specific mechanism that is independent of host response and secondary pathologies. MDPs are composed of a fluorescent dye conjugated to maltohexaose, and are rapidly internalized through the bacteria-specific maltodextrin transport pathway8,9,10,11, endowing the MDPs with a unique combination of high sensitivity and specificity for bacteria. Here, we show that MDPs selectively accumulate within bacteria at millimolar concentrations, and are a thousand-fold more specific for bacteria than mammalian cells. Furthermore, we demonstrate that MDPs can image as few as 105 colony-forming units in vivo and can discriminate between active bacteria and inflammation induced by either lipopolysaccharides or metabolically inactive bacteria.

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Figure 1: In vivo detection of bacteria with MDPs.
Figure 2: Synthesis of MDP-1 and MDP-2.
Figure 3: MDPs have specificity for planktonic bacteria and bacterial biofilms.
Figure 4: MDP-2 images bacteria in vivo.
Figure 5: MDP-2 images bacteria in vivo using internalization through the maltodextrin transporter.

References

  1. Bettegowda, C. et al. Imaging bacterial infections with radiolabeled 1-(2′-deoxy-2′-fluoro-β-D-arabinofuranosyl)-5-iodouracil. Proc. Natl Acad. Sci. USA 102, 1145–1150 (2005).

    CAS  Article  Google Scholar 

  2. Leevy, W. M. et al. Optical imaging of bacterial infection in living mice using a fluorescent near-infrared molecular probe. J. Am. Chem. Soc. 128, 16476–16477 (2006).

    CAS  Article  Google Scholar 

  3. Smith, B. A. et al. Optical imaging of mammary and prostate tumors in living animals using a synthetic near infrared zinc(II)-dipicolylamine probe for anionic cell surfaces. J. Am. Chem. Soc. 132, 67–69 (2010).

    CAS  Article  Google Scholar 

  4. Welling, M. M., Paulusma-Annema, A., Balter, H. S., Pauwels, E. K. & Nibbering, P. H. Technetium-99m labelled antimicrobial peptides discriminate between bacterial infections and sterile inflammations. Eur. J. Nucl. Med. 27, 292–301 (2000).

    CAS  Article  Google Scholar 

  5. Mahfouz, T. et al. 18F-fluorodeoxyglucose positron emission tomography contributes to the diagnosis and management of infections in patients with multiple myeloma: A study of 165 infectious episodes. J. Clin. Oncol. 23, 7857–7863 (2005).

    CAS  Article  Google Scholar 

  6. Leevy, W. M. et al. Noninvasive optical imaging of Staphylococcus aureus bacterial infection in living mice using a Bis-dipicolylamine-Zinc(II) affinity group conjugated to a near-infrared fluorophore. Bioconjug. Chem. 19, 686–692 (2008).

    CAS  Article  Google Scholar 

  7. Rouzet, F. et al. Technetium 99m-labeled annexin V scintigraphy of platelet activation in vegetations of experimental endocarditis. Circulation 117, 781–789 (2008).

    Article  Google Scholar 

  8. Boos, W. & Shuman, H. Maltose/maltodextrin system of Escherichia coli: Transport, metabolism, and regulation. Microbiol. Mol. Biol. Rev. 62, 204–229 (1998).

    CAS  Google Scholar 

  9. Gopal, S. et al. Maltose and maltodextrin utilization by Listeria monocytogenes depend on an inducible ABC transporter which is repressed by glucose. PLoS ONE 5, e10349 (2010).

    Article  Google Scholar 

  10. Oldham, M. L., Khare, D., Quiocho, F. A., Davidson, A. L. & Chen, J. Crystal structure of a catalytic intermediate of the maltose transporter. Nature 450, 515–521 (2007).

    CAS  Article  Google Scholar 

  11. Brass, J. M., Bauer, K., Ehmann, U. & Boos, W. Maltose-binding protein does not modulate the activity of maltoporin as a general porin in Escherichia coli. J. Bacteriol. 161, 720–726 (1985).

    CAS  Google Scholar 

  12. Lipsky, B. A., Itani, K. & Norden, C. Treating foot infections in diabetic patients: A randomized, multicenter, open-label trial of linezolid versus ampicillin-sulbactam/amoxicillin-clavulanate. Clin. Infect. Dis. 38, 17–24 (2004).

    CAS  Article  Google Scholar 

  13. Reiber, G. E., Pecoraro, R. E. & Koepsell, T. D. Risk factors for amputation in patients with diabetes mellitus. A case-control study. Ann. Intern. Med. 117, 97–105 (1992).

    CAS  Article  Google Scholar 

  14. Moore, E. H. Atypical mycobacterial infection in the lung: CT appearance. Radiology 187, 777–782 (1993).

    CAS  Article  Google Scholar 

  15. Erasmus, J. J., McAdams, H. P., Farrell, M. A. & Patz, E. F. Jr Pulmonary nontuberculous mycobacterial infection: Radiologic manifestations. Radiographics 19, 1487–1505 (1999).

    CAS  Article  Google Scholar 

  16. Dahl, M. K. & Manson, M. D. Interspecific reconstitution of maltose transport and chemotaxis in Escherichia coli with maltose-binding protein from various enteric bacteria. J. Bacteriol. 164, 1057–1063 (1985).

    CAS  Google Scholar 

  17. Reuss, R. et al. Intracellular delivery of carbohydrates into mammalian cells through swelling-activated pathways. J. Membr. Biol. 200, 67–81 (2004).

    CAS  Article  Google Scholar 

  18. Line, B. R., Weber, P. B., Lukasiewicz, R. & Dansereau, R. N. Reduction of background activity through radiolabeling of antifibrin Fab′ with 99mTc-dextran. J. Nucl. Med. 41, 1264–1270 (2000).

    CAS  Google Scholar 

  19. Demko, Z. P. & Sharpless, K. B. A click chemistry approach to tetrazoles by Huisgen 1,3-dipolar cycloaddition: Synthesis of 5-acyltetrazoles from azides and acyl cyanides. Angew. Chem. Int. Ed. Engl. 41, 2113–2116 (2002).

    CAS  Article  Google Scholar 

  20. Tornoe, C. W., Christensen, C. & Meldal, M. Peptidotriazoles on solid phase: [1,2,3]-triazoles by regiospecific copper(I)-catalyzed 1,3-dipolar cycloadditions of terminal alkynes to azides. J. Org. Chem. 67, 3057–3064 (2002).

    CAS  Article  Google Scholar 

  21. Dippel, R. & Boos, W. The maltodextrin system of Escherichia coli: Metabolism and transport. J. Bacteriol. 187, 8322–8331 (2005).

    CAS  Article  Google Scholar 

  22. Freundlieb, S., Ehmann, U. & Boos, W. Facilitated diffusion of p-nitrophenyl-alpha-D-maltohexaoside through the outer membrane of Escherichia coli. Characterization of LamB as a specific and saturable channel for maltooligosaccharides. J. Biol. Chem. 263, 314–320 (1988).

    CAS  Google Scholar 

  23. Baba, T. et al. Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection. Mol. Syst. Biol. 2, 2006.0008 (2006).

    Article  Google Scholar 

  24. Reid, G. Biofilms in infectious disease and on medical devices. Int. J. Antimicrob. Agents 11, 223–226 (1999).

    CAS  Article  Google Scholar 

  25. Author, A. N. Panel discussion on biofilms in urinary tract infection. Int. J. Antimicrob. Agents 11, 237–239 (1999).

    Article  Google Scholar 

  26. Hall-Stoodley, L., Costerton, J. W. & Stoodley, P. Bacterial biofilms: From the natural environment to infectious diseases. Nature Rev. Microbiol. 2, 95–108 (2004).

    CAS  Article  Google Scholar 

  27. Kolodkin-Gal, I. et al. D-amino acids trigger biofilm disassembly. Science 328, 627–629 (2010).

    CAS  Article  Google Scholar 

  28. Dehoux, M. J., van Beneden, R. P., Fernandez-Celemin, L., Lause, P. L. & Thissen, J. P. Induction of MafBx and Murf ubiquitin ligase mRNAs in rat skeletal muscle after LPS injection. FEBS Lett. 544, 214–217 (2003).

    CAS  Article  Google Scholar 

  29. Luo, G., Niesel, D. W., Shaban, R. A., Grimm, E. A. & Klimpel, G. R. Tumor necrosis factor alpha binding to bacteria: evidence for a high-affinity receptor and alteration of bacterial virulence properties. Infect. Immun. 61, 830–835 (1993).

    CAS  Google Scholar 

  30. Larson, T. J., Ludtke, D. N. & Bell, R. M. sn-Glycerol-3-phosphate auxotrophy of plsB strains of Escherichia coli: evidence that a second mutation, plsX, is required. J. Bacteriol. 160, 711–717 (1984).

    CAS  Google Scholar 

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Acknowledgements

This project has been funded in whole or in part with Federal funds from the National Heart, Lung, and Blood Institute, National Institutes of Health, Department of Health and Human Services, under Contract No. HHSN268201000043C, NSF-BES-0546962 Career Award (N.M.) and NIH RO1 HL096796-01 (N.M.).

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Authors

Contributions

X.N. synthesized and characterized MDP-1 and MDP-2, designed and analysed experiments, and wrote the manuscript. S.L. designed, carried out and analysed experiments, and contributed to the writing of the manuscript. Z.W. performed MS experiments to characterize all intermediates and final products and proof read the manuscript. D.K. carried out in vitro experiments. B.S. prepared biofilms and performed confocal laser scanning microscopy. E.G. supervised the preparation of biofilms and proof read the manuscript. N.M. designed and supervised the project and contributed to the writing of the manuscript.

Corresponding authors

Correspondence to Seungjun Lee or Niren Murthy.

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The authors declare no competing financial interests.

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Ning, X., Lee, S., Wang, Z. et al. Maltodextrin-based imaging probes detect bacteria in vivo with high sensitivity and specificity. Nature Mater 10, 602–607 (2011). https://doi.org/10.1038/nmat3074

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