A cardiomyocyte mannose receptor system is involved in Trypanosoma cruzi invasion and is down-modulated after infection.
Mannosyl binding sites were detected "in vitro" on cardiomyocytes (CM) surface using horseradish peroxidase (HRP) as the ligand. Binding assays revealed a specific recognition system, which was time- and concentration-dependent. The binding required physiological pH and was inhibited by EDTA and trypsin treatments. HRP binding was reduced by pre-incubations with low concentrations of D-mannose. Ultrastructural analysis of the endocytic process was followed using HRP coupled to colloidal gold particles (HRP-Au). The tracer was found within caveolae characterizing early steps of the receptor-mediated endocytosis. The addition of 10 mM D-mannose to the interaction medium blocked Trypanosoma cruzi uptake by CM. The labeling of CM with a subsaturating concentration of HRP-Au before their infection showed, by ultrastructural studies, that its association with trypomastigote forms occurred frequently near to HRP-gold particles that could also be seen to comprise the parasitophorous vacuole. After infection of CM with T. cruzi, a considerable reduction on HRP binding was noticed. Binding was almost completely restored by treating the infected cultures with the trypanocidal drug Nifurtimox. Our "in vitro" findings suggest that cardiomyocyte's mannose receptors localized at the sarcolemma mediates T. cruzi recognition and can be down-modulated by parasite infection.
Soeiro
M de N
Mde N
Lab. de Biologia Celular, Depto. de Ultra-estrutura e Biologia Celular, Instituto Oswaldo Cruz, Rio de Janeiro, Brazil.
Paiva
M M
MM
Barbosa
H S
HS
Meirelles
M de N
Mde N
Araújo-Jorge
T C
TC
D000818:Animals; D015536:Down-Regulation; D004705:Endocytosis; D004797:Enzyme-Linked Immunosorbent Assay; D005690:Galactose / Q000378:metabolism; D006321:Heart / Q000196:embryology; D006735:Horseradish Peroxidase / Q000378:metabolism / Q000648:ultrastructure; D006863:Hydrogen-Ion Concentration; D037181:Lectins, C-Type; D008358:Mannose / Q000378:metabolism; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins; D051379:Mice; D008854:Microscopy, Electron; D009206:Myocardium / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000378:metabolism*; D014349:Trypanosoma cruzi / Q000378:metabolism*; D014352:Trypanosomiasis / Q000378:metabolism* / Q000469:parasitology
D037181:Lectins, C-Type; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins; D011956:Receptors, Cell Surface; D006735:Horseradish Peroxidase; D008358:Mannose; D005690:Galactose