Trypanosoma cruzi surface mucins are involved in the attachment to the Triatoma infestans rectal ampoule.
Camara, M. L. M., Balouz, V., Centeno Camean, C., Cori, C. R., Kashiwagi, G. A., Gil, S. A., Macchiaverna, N. P., Cardinal, M. V., Guaimas, F., Lobo, M. M., de Lederkremer, R. M., Gallo-Rodriguez, C. and Buscaglia, C. A.
Instituto de Investigaciones Biotecnologicas-Instituto Tecnologico de Chascomus (IIB-INTECh), Universidad Nacional de San Martin (UNSAM) and Consejo Nacional de investigaciones cientificas y tecnicas (CONICET), Buenos Aires, Argentina.
Universidad de Buenos Aires, Facultad de Ciencias Exactas y Naturales, Departamento de Quimica Organica, Pabellon 2, Ciudad Universitaria, C1428EGA Buenos Aires, Argentina.
CONICET-UBA, Centro de Investigacion en Hidratos de Carbono (CIHIDECAR), C1428EGA Buenos Aires, Argentina.
Laboratorio de Eco-Epidemiologia, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires e Instituto de Ecologia, Genetica y Evolucion de Buenos Aires (IEGEBA), UBA-CONICET, C1428EGA Buenos Aires, Argentina.
BACKGROUND: Trypanosoma cruzi, the agent of Chagas disease, is a protozoan parasite transmitted to humans by blood-sucking triatomine vectors. However, and despite its utmost biological and epidemiological relevance, T. cruzi development inside the digestive tract of the insect remains a poorly understood process. METHODS/PRINCIPLE FINDINGS: Here we showed that Gp35/50 kDa mucins, the major surface glycoproteins from T. cruzi insect-dwelling forms, are involved in parasite attachment to the internal cuticle of the triatomine rectal ampoule, a critical step leading to its differentiation into mammal-infective forms. Experimental evidence supporting this conclusion could be summarized as follows: i) native and recombinant Gp35/50 kDa mucins directly interacted with hindgut tissues from Triatoma infestans, as assessed by indirect immunofluorescence assays; ii) transgenic epimastigotes over-expressing Gp35/50 kDa mucins on their surface coat exhibited improved attachment rates (~2-3 fold) to such tissues as compared to appropriate transgenic controls and/or wild-type counterparts; and iii) certain chemically synthesized compounds derived from Gp35/50 kDa mucins were able to specifically interfere with epimastigote attachment to the inner lining of T. infestans rectal ampoules in ex vivo binding assays, most likely by competing with or directly blocking insect receptor(s). A solvent-exposed peptide (smugS peptide) from the Gp35/50 kDa mucins protein scaffolds and a branched, Galf-containing trisaccharide (Galfbeta1-4[Galpbeta1-6]GlcNAcalpha) from their O-linked glycans were identified as main adhesion determinants for these molecules. Interestingly, exogenous addition of a synthetic Galfbeta1-4[Galpbeta1-6]GlcNAcalpha derivative or of oligosaccharides containing this structure impaired the attachment of Dm28c but not of CL Brener epimastigotes to triatomine hindgut tissues; which correlates with the presence of Galf residues on the Gp35/50 kDa mucins' O-glycans on the former but not the latter parasite clone. CONCLUSION/SIGNIFICANCE: These results provide novel insights into the mechanisms underlying T. cruzi-triatomine interplay, and indicate that inter-strain variations in the O-glycosylation of Gp35/50 kDa mucins may lead to differences in parasite differentiation and hence, in parasite transmissibility to the mammalian host. Most importantly, our findings point to Gp35/50 kDa mucins and/or the Galf biosynthetic pathway, which is absent in mammals and insects, as appealing targets for the development of T. cruzi transmission-blocking strategies.
PLoS Neglected Tropical Diseases 13(5): e0007418 (2019)