Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

APOLs with low pH dependence can kill all African trypanosomes

Abstract

The primate-specific serum protein apolipoprotein L1 (APOL1) is the only secreted member of a family of cell death promoting proteins1,2,3,4. APOL1 kills the bloodstream parasite Trypanosoma brucei brucei, but not the human sleeping sickness agents T.b. rhodesiense and T.b. gambiense 3. We considered the possibility that intracellular members of the APOL1 family, against which extracellular trypanosomes could not have evolved resistance, could kill pathogenic T. brucei subspecies. Here we show that recombinant APOL3 (rAPOL3) kills all African trypanosomes, including T.b. rhodesiense, T.b. gambiense and the animal pathogens Trypanosoma evansi, Trypanosoma congolense and Trypanosoma vivax. However, rAPOL3 did not kill more distant trypanosomes such as Trypanosoma theileri or Trypanosoma cruzi. This trypanolytic potential was partially shared by rAPOL1 from Papio papio (rPpAPOL1). The differential killing ability of rAPOL3 and rAPOL1 was associated with a distinct dependence on acidic pH for activity. Due both to its instability and toxicity when injected into mice, rAPOL3 cannot be used for the treatment of infection, but an experimental rPpAPOL1 mutant inspired by APOL3 exhibited enhanced trypanolytic activity in vitro and the ability to completely inhibit T.b. gambiense infection in mice. We conclude that pH dependence influences the trypanolytic potential of rAPOLs.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: In vitro trypanolytic potential of various rAPOLs.
Fig. 2: Phenotype of rAPOL3-mediated trypanolysis.
Fig. 3: Differential pH dependence of rAPOL3, rAPOL1 and rPpAPOL1.
Fig. 4: In vivo trypanolytic potential of various rAPOLs.

Similar content being viewed by others

References

  1. Vanhollebeke, B. & Pays, E. The function of apolipoproteins L. Cell. Mol. Life Sci. 63, 1937–1944 (2006).

    Article  CAS  PubMed  Google Scholar 

  2. Hu, C. A., Klopfer, E. I. & Ray, P. E. Human apolipoprotein L1 (ApoL1) in cancer and chronic kidney disease. FEBS Lett. 586, 947–955 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Pays, E., Vanhollebeke, B., Uzureau, P., Lecordier, L. & Pérez-Morga, D. The molecular arms race between African trypanosomes and humans. Nat. Rev. Microbiol. 12, 575–584 (2014).

    Article  CAS  PubMed  Google Scholar 

  4. Uzureau, S. et al. Apolipoproteins L control cell death triggered by TLR3/TRIF signaling in dendritic cells. Eur. J. Immunol. 46, 1854–1866 (2016).

    Article  CAS  PubMed  Google Scholar 

  5. Vanhamme, L. et al. Apolipoprotein L-I is the trypanosome lytic factor of human serum. Nature 422, 83–87 (2003).

    Article  CAS  PubMed  Google Scholar 

  6. Pérez-Morga, D. et al. Apolipoprotein L-I promotes trypanosome lysis by forming pores in lysosomal membranes. Science 309, 469–472 (2005).

    Article  PubMed  Google Scholar 

  7. Vanwalleghem, G. et al. Coupling of lysosomal and mitochondrial membrane permeabilization in trypanolysis by APOL1. Nat. Commun. 6, 8078 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Greene, A. S. & Hajduk, S. L. Trypanosome lytic factor-1 initiates oxidation-stimulated osmotic lysis of Trypanosoma brucei brucei. J. Biol. Chem. 291, 3063–3075 (2016).

    Article  CAS  PubMed  Google Scholar 

  9. Shimamura, M., Hager, K. M. & Hajduk, S. L. The lysosomal targeting and intracellular metabolism of trypanosome lytic factor by Trypanosoma brucei brucei. Mol. Biochem. Parasitol. 115, 227–237 (2001).

    Article  CAS  PubMed  Google Scholar 

  10. Lecordier, L. et al. Identification of Trypanosoma brucei components involved in trypanolysis by normal human serum. Mol. Microbiol. 94, 625–636 (2014).

    Article  CAS  PubMed  Google Scholar 

  11. Thomson, R. & Finkelstein, A. Human trypanolytic factor APOL1 forms pH-gated cation-selective channels in planar lipid bilayers: relevance to trypanosome lysis. Proc. Natl Acad. Sci. USA 112, 2894–2899 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Alsford, S., Currier, R. B., Guerra-Assunção, J. A., Clark, T. G. & Horn, D. Cathepsin-L can resist lysis by human serum in Trypanosoma brucei brucei. PLoS Pathog. 10, e1004130 (2014).

    Article  PubMed  PubMed Central  Google Scholar 

  13. Xong, H. V. et al. A VSG expression site-associated gene confers resistance to human serum in Trypanosoma rhodesiense. Cell 95, 839–846 (1998).

    Article  CAS  PubMed  Google Scholar 

  14. Uzureau, P. et al. Mechanism of Trypanosoma gambiense resistance to human serum. Nature 501, 430–434 (2013).

    Article  CAS  PubMed  Google Scholar 

  15. Capewell, P. et al. The TgsGP gene is essential for resistance to human serum in Trypanosoma brucei gambiense. PLoS Pathog. 9, e1003686 (2013).

    Article  PubMed  PubMed Central  Google Scholar 

  16. Lecordier, L. et al. C-terminal mutants of apolipoprotein L-I efficiently kill both Trypanosoma brucei brucei and Trypanosoma brucei rhodesiense. PLoS Pathog. 5, e1000685 (2009).

    Article  PubMed  PubMed Central  Google Scholar 

  17. Genovese, G. et al. Association of trypanolytic apoL1 variants with kidney disease in African-Americans. Science 329, 841–845 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Thomson, R., Molina-Portela, P., Mott, H., Carrington, M. & Raper, J. Hydrodynamic gene delivery of baboon trypanosome lytic factor eliminates both animal and human-infective African trypanosomes. Proc. Natl Acad. Sci. USA 106, 19509–19514 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Page, N. M., Butlin, D. J., Lomthaisong, K. & Lowry, P. J. The human apolipoprotein L gene cluster: identification, classification, and sites of distribution. Genomics 74, 71–78 (2001).

    Article  CAS  PubMed  Google Scholar 

  20. Bruggeman, L. A. et al. Plasma apolipoprotein L1 levels do not correlate with CKD. J. Am. Soc. Nephrol. 25, 634–644 (2014).

    Article  CAS  PubMed  Google Scholar 

  21. Vanhollebeke, B., Lecordier, L., Perez-Morga, D., Amiguet-Vercher, A. & Pays, E. Human serum lyses Trypanosoma brucei by triggering uncontrolled swelling of the parasite lysosome. J. Eukaryot. Microbiol. 54, 448–451 (2007).

    Article  PubMed  Google Scholar 

  22. Pays, E. et al. The trypanolytic factor of human serum. Nat. Rev. Microbiol. 4, 477–486 (2006).

    Article  CAS  PubMed  Google Scholar 

  23. Dutoya, S. et al. A novel C-terminal kinesin is essential for maintaining functional acidocalcisomes in Trypanosoma brucei. J. Biol. Chem. 276, 49117–49124 (2001).

    Article  CAS  PubMed  Google Scholar 

  24. Barrera, F. N. et al. Roles of carboxyl groups in the transmembrane insertion of peptides. J. Mol. Biol. 413, 359–371 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Willyard, C. Putting sleeping sickness to bed. Nat. Med. 17, 14–17 (2011).

    Article  CAS  PubMed  Google Scholar 

  26. Raper, J. & Friedman, D. J. Parasitology: molecular one-upmanship. Nature 501, 322–323 (2013).

    Article  CAS  PubMed  Google Scholar 

  27. Kageruka, P. et al. Infectivity of Trypanosoma (Trypanozoon) brucei gambiense for baboons (Papio hamadryas, Papio papio). Ann. Soc. Belg. Med. Trop. 71, 39–46 (1991).

    CAS  PubMed  Google Scholar 

  28. Cooper, A. et al. A primate APOL1 variant that kills Trypanosoma brucei gambiense. PLoS Negl. Trop. Dis. 10, e0004903 (2016).

    Article  PubMed  PubMed Central  Google Scholar 

  29. Radwanska, M. et al. Novel primer sequences for a polymerase chain reaction-based detection of Trypanosoma brucei gambiense. Am. J. Trop. Med. Hyg. 67, 289–295 (2002).

    Article  CAS  PubMed  Google Scholar 

  30. Pyana Pati, P. et al. Melarsoprol sensitivity profile of Trypanosoma brucei gambiense isolates from cured and relapsed sleeping sickness patients from the Democratic Republic of the Congo. PLoS Negl. Trop. Dis. 8, e3212 (2014).

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

The authors thank A. Kremer (Ghent) for the FIB–SEM acquisitions. This work was supported by the European Research Council (ERC 669007-APOLs), the Interuniversity Attraction Poles Programme–Belgian Science Policy (PAI P7-41) and the PDR-FNRS (PDR T.0159.13). The Center for Microscopy and Molecular Imaging is supported by the European Regional Development Fund and Wallonia.

Author information

Authors and Affiliations

Authors

Contributions

E.P., F.F. and D.P.M. designed the research. F.F., L.L., G.V., P.U., M.S. and P.T. performed the research. B.V. supervised some experiments. N.V.R. and P.B. adapted the trypanosome strains to in vitro growth. E.P. and F.F. wrote the paper.

Corresponding author

Correspondence to Etienne Pays.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Supplementary Information

Supplementary Figures 1–12, Supplementary Table 1

Life Sciences Reporting Summary

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Fontaine, F., Lecordier, L., Vanwalleghem, G. et al. APOLs with low pH dependence can kill all African trypanosomes. Nat Microbiol 2, 1500–1506 (2017). https://doi.org/10.1038/s41564-017-0034-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41564-017-0034-1

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing