title: Rab5 activation by Toll-like receptor 2 is required for Trypanosoma cruzi internalization and replication in macrophages.
abstract: Trypanosoma cruzi can infect and replicate in macrophages. During invasion, T. cruzi interacts with different macrophage receptors to induce its own phagocytosis. However, the nature of those receptors and the molecular mechanisms involved are poorly understood. In this study, we demonstrate that T. cruzi metacyclic trypomastigotes but not epimastigotes were able to induce Rab5 activation and binding to the early endosomes in peritoneal macrophages. In this process, active Rab5 colocalized with parasites in the phagosome and with the Rab5A effector molecule early endosomal antigen 1. Phagosome formation and T. cruzi internalization were inhibited in Raw 264.7 macrophages expressing a dominant-negative form of Rab5 [(S34N)Rab5]. Using T. cruzi membrane extracts, we verified that the Rab5 activation depends on the interaction between parasite surface molecules and macrophages surface molecule. In addition, during infection of macrophages, phosphatidylinositol 3-kinase (PI3K) pathway was activated. Assays carried out using a selective PI3K inhibitor (LY294002) showed that the PI3K activation is essential for Rab5 activation by T. cruzi infection and for the entrance and intracellular replication of T. cruzi in macrophages. Moreover, using macrophages from knockout mice, we found that activation of Rab5, fusion of early endosomes and phagocytosis induced by T. cruzi infection involved Toll-like receptor (TLR)2 but were independent of TLR4 receptors.
authors.affiliation: Centro de Biología Molecular Severo Ochoa, CSIC-UAM, Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain.
authors.forename: Elena
authors.initials: E
authors.lastname: Maganto-Garcia
authors.forename: Carmen
authors.initials: C
authors.lastname: Punzon
authors.forename: Cox
authors.initials: C
authors.lastname: Terhorst
authors.forename: Manuel
authors.initials: M
authors.lastname: Fresno
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D011992:Endosomes / Q000378:metabolism; D005260:Female; D008264:Macrophages / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D018345:Mice, Knockout; D008856:Microscopy, Fluorescence; D010588:Phagosomes / Q000378:metabolism; D051195:Toll-Like Receptor 2 / Q000737:chemistry*; D051197:Toll-Like Receptor 4 / Q000378:metabolism; D014349:Trypanosoma cruzi / Q000378:metabolism*; D014352:Trypanosomiasis / Q000378:metabolism* / Q000469:parasitology*; D020696:rab5 GTP-Binding Proteins / Q000737:chemistry*
chemical_list: D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D020696:rab5 GTP-Binding Proteins