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Real‐time imaging of Leishmania mexicana ‐infected early phagosomes: a study using primary macrophages generated from green fluorescent protein‐Rab5 transgenic mice
Author(s) -
Lippuner Christoph,
Paape Daniel,
Paterou Athina,
Brand Janko,
Richardson Melville,
Smith Andrew J.,
Hoffmann Kirstin,
Brinkmann Volker,
Blackburn Clare,
Aebischer Toni
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.08-108712
Subject(s) - phagosome , leishmania , microbiology and biotechnology , endosome , lipophosphoglycan , biology , leishmania mexicana , green fluorescent protein , intracellular parasite , genetically modified mouse , leishmania major , phagocytosis , intracellular , transgene , parasite hosting , gene , biochemistry , world wide web , computer science
The small GTPase Rab5 is a key regulator of endosome/phagosome maturation and in intravesicular infections marks a phagosome stage at which decisions over pathogen replication or destruction are integrated. It is currently unclear whether Leishmania‐ in fected phagosomes uniformly pass through a Rab5 + stage on their intracellular path to compartments with late endosomal/ early lysosomal characteristics. Differences in routes and final compartments could have consequences for accessibility to antileishmanial drugs. Here, we generated a unique gfp‐rab5 transgenic mouse model to visualize Rab5 recruitment to early parasite‐containing phagosomes in primary host cells. Using real‐time fluorescence imaging of phagosomes carrying Leishmania mexicana , we determined that parasite‐infested phagosomes follow a uniform sequence of transient Rab5 recruitment. Residence in Rab5 + compartments was much shorter compared with phagosomes harboring latex beads. Furthermore, a comparative analysis of parasite life‐cycle stages and mutants deficient in lpg1 , the gene encoding the enzyme required for synthesis of the dominant surface lipophosphoglycan, indicated that parasite surface ligands and host cell receptors modulate pathogen residence times in Rab5 + phagosomes, but, as far as tested, had no significant effect on intracellular L. mexicana survival or replication.—Lippuner, C, Paape, D., Paterou, A., Brand, J., Richardson, M., Smith, A. J., Hoffmann, K., Brinkmann, V., Blackburn, C, Aebischer, T. Real‐time imaging of Leishmania mexicana‐ in fected early phagosomes: a study using primary macrophages generated from green fluorescent protein‐Rab5 transgenic mice. FASEB J . 23, 483–491 (2009)