Intracellular Survival of Leishmania major Depends on Uptake and Degradation of Extracellular Matrix Glycosaminoglycans by Macrophages

Thomas Naderer; Joanne Heng; Eleanor C Saunders; Joachim Kloehn; Thusitha W Rupasinghe; Tracey J Brown; Malcolm J McConville

doi:10.1371/journal.ppat.1005136

Intracellular Survival of Leishmania major Depends on Uptake and Degradation of Extracellular Matrix Glycosaminoglycans by Macrophages

PLoS Pathog. 2015 Sep 3;11(9):e1005136. doi: 10.1371/journal.ppat.1005136. eCollection 2015 Sep.

Authors

Thomas Naderer¹, Joanne Heng², Eleanor C Saunders², Joachim Kloehn², Thusitha W Rupasinghe³, Tracey J Brown⁴, Malcolm J McConville²

Affiliations

¹ The Department of Biochemistry and Molecular Biology and the Bio21 Institute of Molecular Science and Biotechnology, University of Melbourne, Parkville, Victoria, Australia; Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia.
² The Department of Biochemistry and Molecular Biology and the Bio21 Institute of Molecular Science and Biotechnology, University of Melbourne, Parkville, Victoria, Australia.
³ Metabolomics Australia, Bio21 Institute of Molecular Science and Biotechnology, University of Melbourne, Parkville, Victoria, Australia.
⁴ Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia.

Abstract

Leishmania parasites replicate within the phagolysosome compartment of mammalian macrophages. Although Leishmania depend on sugars as a major carbon source during infections, the nutrient composition of the phagolysosome remains poorly described. To determine the origin of the sugar carbon source in macrophage phagolysosomes, we have generated a N-acetylglucosamine acetyltransferase (GNAT) deficient Leishmania major mutant (∆gnat) that is auxotrophic for the amino sugar, N-acetylglucosamine (GlcNAc). This mutant was unable to grow or survive in ex vivo infected macrophages even when macrophages were cultivated in presence of exogenous GlcNAc. In contrast, the L. major ∆gnat mutant induced normal skin lesions in mice, suggesting that these parasites have access to GlcNAc in tissue macrophages. Intracellular growth of the mutant in ex vivo infected macrophages was restored by supplementation of the macrophage medium with hyaluronan, a GlcNAc-rich extracellular matrix glycosaminoglycan. Hyaluronan is present and constitutively turned-over in Leishmania-induced skin lesions and is efficiently internalized into Leishmania containing phagolysosomes. These findings suggest that the constitutive internalization and degradation of host glycosaminoglycans by macrophages provides Leishmania with essential carbon sources, creating a uniquely favorable niche for these parasites.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Acetylglucosamine / metabolism
Acetyltransferases / genetics
Acetyltransferases / metabolism
Animals
Cell Survival
Cells, Cultured
Extracellular Matrix / immunology
Extracellular Matrix / metabolism*
Extracellular Matrix / pathology
Gene Deletion
Glycosaminoglycans / metabolism*
Host-Parasite Interactions*
Hydrolysis
Kinetics
Leishmania major / genetics
Leishmania major / growth & development
Leishmania major / immunology
Leishmania major / physiology*
Leishmania mexicana / genetics
Leishmania mexicana / growth & development
Leishmania mexicana / immunology
Leishmania mexicana / physiology
Leishmaniasis, Cutaneous / immunology
Leishmaniasis, Cutaneous / metabolism
Leishmaniasis, Cutaneous / parasitology
Leishmaniasis, Cutaneous / pathology
Lysosomes / immunology
Lysosomes / metabolism
Lysosomes / parasitology*
Lysosomes / pathology
Macrophages / immunology
Macrophages / metabolism
Macrophages / parasitology*
Macrophages / pathology
Male
Mice, Inbred BALB C
Phagocytosis*
Protozoan Proteins / genetics
Protozoan Proteins / metabolism
Species Specificity
Specific Pathogen-Free Organisms

Substances

Glycosaminoglycans
Protozoan Proteins
Acetyltransferases
glucosamine acetyltransferase
Acetylglucosamine

Grants and funding

This work was supported by the National Health and Medical Research Council, Canberra, Australia (www.nhmrc.gov.au): Project Grant 1024839 (TN) and Project Grant 1006023 (MJM). MJM is a NHMRC Principal Research Fellow (566643). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.