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In vivo tropism of Salmonella Typhi toxin to cells expressing a multiantennal glycan receptor

Abstract

Typhoid fever is a life-threatening disease, but little is known about the molecular bases for its unique clinical presentation. Typhoid toxin, a unique virulence factor of Salmonella Typhi (the cause of typhoid fever), recapitulates in an animal model many symptoms of typhoid fever. Typhoid toxin binding to its glycan receptor Neu5Ac is central, but, due to the ubiquity of Neu5Ac, how typhoid toxin causes specific symptoms remains elusive. Here we show that typhoid toxin displays in vivo tropism to cells expressing multiantennal glycoprotein receptors, particularly on endothelial cells of arterioles in the brain and immune cells, which is in line with typhoid symptoms. Neu5Ac displayed by multiantennal N-glycans, rather than a single Neu5Ac, appears to serve as the high-affinity receptor, as typhoid toxin possesses five identical binding pockets per toxin. Human counterparts also express the multiantennal Neu5Ac receptor. Here we also show that mice immunized with inactive typhoid toxins and challenged with wild-type typhoid toxin presented neither the characteristic in vivo tropism nor symptoms. These mice were protected against a lethal-dose toxin challenge, but Ty21a-vaccinated mice were not. Cumulatively, these results reveal remarkable features describing how a bacterial exotoxin induces virulence exclusively in specific cells at the organismal level.

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Change history

  • 13 June 2019

    An amendment to this paper has been published and can be accessed via a link at the top of the paper.

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Acknowledgements

The authors thank J. E. Galán for his suggestion on the typhoid toxoid vaccination study, which was initiated while J.S. was at Yale, N. Nishimura for her comments on brain images, and X. Gao for plasmid pET21a-pltB-His6. This work was supported by a Cornell University Startup, PCCW Affinito-Stewart Award and the USDA/NIFA Hatch project 1010701 (to J.S.), the NIH, NCI, under contract no. HHSN261200800001E (to R.N.), the NIH, NIDCD intramural research programme (to J.Z.), and NIH R01 AI114730 to J.C.P. The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Author information

Y.-A.Y. conducted the experiments shown in Figs. 1, 2, 3a and 4d,g–k, Supplementary Figs. 1–6 and 9–11 and Supplementary Videos 1–10. S.L. conducted the experiments in Figs. 4a–c,e,f. J.Z. conducted the analyses and simulations in Figs. 3b–e. R.N. supervised the computational project. L.D. contributed to Fig. 3. A.J.T. conducted the experiments in Supplementary Fig. 8. R.M. conducted experiments in Fig. 3f and Supplementary Fig. 7. B.T. contributed to Supplementary Figs. 7 and 8. J.C.P. supervised the glycan microarray project. J.S. was involved in the design, interpretation and supervision of this study. Y.-A.Y. and J.S. wrote the paper and all the authors commented on the manuscript.

Competing interests

The authors declare no competing financial interests.

Correspondence to Jeongmin Song.

Electronic supplementary material

  1. Supplementary Information

    Supplementary Figures 1–11, Supplementary Tables 1-4, Supplementary Methods, Supplementary References, Supplementary Data

  2. Life Sciences Reporting Summary

  3. Supplementary Table 5

    Related to Fig. 3f and Supplementary Fig. 7. Lists of glycans and glycan microarray results. Data for individual glycans are reported as relative fluorescence intensities from four of six replicates (after removal of highest and lowest values)

  4. Supplementary Video 1

    Related to Fig. 1c. A representative video showing ataxia signs typically seen 4.5–5 days after WT toxin administration

  5. Supplementary Video 2

    Related to Fig. 1c. A representative video showing clonic seizure signs typically seen 5–6 days after WT toxin administration

  6. Supplementary Video 3

    Related to Fig. 1d. A representative balance beam test of C57BL/6 mice administered PBS. Balance beam tests were conducted 5 days after PBS buffer administration

  7. Supplementary Video 4

    Related to Fig. 1d. A representative result of balance beam tests of mice administered WT toxin. Balance beam tests were conducted 5 days after WT toxin administration

  8. Supplementary Video 5

    Related to Fig. 1d. Another representative result of balance beam tests of mice administered WT toxin. Balance beam tests were conducted 5 days after WT toxin administration

  9. Supplementary Video 6

    Related to Fig. 1d. A representative result of balance beam tests of mice administered a PltB catalytic mutant toxin. Balance beam tests were conducted 5 days after indicated toxin administration

  10. Supplementary Video 7

    Related to Fig. 1f. A representative result of balance beam tests of mice administered a PltA catalytic mutant toxin. Balance beam tests were conducted 5 days after indicated toxin administration

  11. Supplementary Video 8

    Related to Fig. 1f. A representative result of balance beam tests of mice administered a CdtB catalytic mutant toxin. Balance beam tests were conducted 5 days after indicated toxin administration

  12. Supplementary Video 9

    Related to Supplementary Fig. 2. A representative result of balance beam tests of mice infected with a lethal dose of S. Typhimurium. A group of C57BL/6 mice were infected orally with 107 S. Typhimurium SL1344. The balance beam test was conducted 5 days after infection

  13. Supplementary Video 10

    Related to Fig. 4i. A representative result of balance beam tests of mice immunized by typhoid toxoid and challenged by a lethal dose of typhoid toxin. A group of C57BL/6 mice were immunized by subcutaneous injections of typhoid toxoid via a standard prime-boost regimen. Two weeks after boost, the immunized mice were challenged with 2 μg WT typhoid toxin. Balance beam tests were conducted 5 days after the WT toxin challenge

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Further reading

Fig. 1: Typhoid toxin causes neurological complications associated with motor function deficits where both PltB and CdtB subunits play critical roles.
Fig. 2: Typhoid toxin has in vivo tropism to endothelial cells of arterioles in the brain and to immune cells.
Fig. 3: Typhoid toxin binds preferentially to Neu5Ac displayed in the context of multiantennal N-glycans over linear N-glycans.
Fig. 4: Vaccination of naive mice with genetically engineered inactive typhoid toxin or PltB subunit alone completely protects the mice against a lethal-dose toxin challenge.