Trypanosoma cruzi heparin-binding proteins mediate the adherence of epimastigotes to the midgut epithelial cells of Rhodnius prolixus.
Heparin-binding proteins (HBPs) have been demonstrated in both infective forms of Trypanosoma cruzi and are involved in the recognition and invasion of mammalian cells. In this study, we evaluated the potential biological function of these proteins during the parasite-vector interaction. HBPs, with molecular masses of 65·8 kDa and 59 kDa, were isolated from epimastigotes by heparin affinity chromatography and identified by biotin-conjugated sulfated glycosaminoglycans (GAGs). Surface plasmon resonance biosensor analysis demonstrated stable receptor-ligand binding based on the association and dissociation values. Pre-incubation of epimastigotes with GAGs led to an inhibition of parasite binding to immobilized heparin. Competition assays were performed to evaluate the role of the HBP-GAG interaction in the recognition and adhesion of epimastigotes to midgut epithelial cells of Rhodnius prolixus. Epithelial cells pre-incubated with HBPs yielded a 3·8-fold inhibition in the adhesion of epimastigotes. The pre-treatment of epimastigotes with heparin, heparan sulfate and chondroitin sulfate significantly inhibited parasite adhesion to midgut epithelial cells, which was confirmed by scanning electron microscopy. We provide evidence that heparin-binding proteins are found on the surface of T. cruzi epimastigotes and demonstrate their key role in the recognition of sulfated GAGs on the surface of midgut epithelial cells of the insect vector.
Laboratório de Ultra-estrutura Celular, Instituto Oswaldo Cruz/FIOCRUZ, RJ, Brazil.
F O R
FO
Oliveira
C R
CR
Alves
F
F
Souza-Silva
C M
CM
Calvet
L M C
LM
Côrtes
M S
MS
Gonzalez
L
L
Toma
R I
RI
Bouças
H B
HB
Nader
M C S
MC
Pereira
D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D004847:Epithelial Cells / Q000469:parasitology*; D041981:Gastrointestinal Tract / Q000166:cytology / Q000469:parasitology; D006493:Heparin / Q000378:metabolism*; D006790:Host-Parasite Interactions; D015800:Protozoan Proteins / Q000378:metabolism / Q000494:pharmacology*; D012239:Rhodnius / Q000469:parasitology*; D014349:Trypanosoma cruzi / Q000254:growth & development / Q000502:physiology*
D015800:Protozoan Proteins; D006493:Heparin