Mem Inst Oswaldo Cruz, Rio de Janeiro, 104 (Suppl.I) July 2009
Original Article

?-N-acetylglucosamine-linked O-glycans of sialoglycoproteins (Tc-mucins) from Trypanosoma cruzi Colombiana strain

Adriane R TodeschiniI; Eliane G de AlmeidaI; Orlando A AgrellosI; Christopher JonesII; José O PreviatoI; Lucia Mendonça-PreviatoI, +

IInstituto de Biofísica Carlos Chagas Filho, Centro de Ciências da Saúde, Universidade Federal do Rio de Janeiro, 21949-900 Rio de Janeiro, RJ, Brasil
IILaboratory for Molecular Structure, National Institute for Biological Standards and Control, South Mimms, Herts, UK

Page: 270-274 DOI: 10.1590/S0074-02762009000900035
3361 views 1019 downloads

Trypanosoma cruzi sialoglycoproteins (Tc-mucins) are mucin-like molecules linked to a parasite membrane via a glycosylphosphatidylinositol anchor. We previously determined the structures of Tc-mucin O-glycan domains from several T. cruzi strains and observed significant differences among them. We now report the amino acid content and structure of Tc-mucin O-glycan chains from T. cruzi Colombiana, a strain resistant to common trypanocidal drugs. Amino acid analysis demonstrated the predominance of threonine residues (42%) and helped to identify the O-glycans as belonging to a Tc-mucin family that contain a u03b2-galactofuranose (u03b2-Galf) residue attached to an u03b1-N-acetylglucosamine (u03b1-GlcNAc) O-4, with the most complex glycan, a pentasaccharide-GlcNAc-ol with a branched trigalactopyranose chain, on the GlcNAc O-6. The presence of u03b2-Galf on O-glycans from T. cruzi Colombiana mucins supports the use of glycosylation as a phylogenetic marker for the classification of Colombiana in the T. cruzi I group.

Chagas disease, caused by the protozoan parasite Trypanosoma cruzi, affects nearly eight million people in South and Central America (WHO 2007) and is becoming a significant health problem in developed countries such as the United States and Spain (Riera et al. 2006, Hotez 2008). The infective forms of T. cruzi, known as trypomastigotes, invade mammalian cells by a not yet completely defined mechanism. However, glycoconjugates on the parasite cell surface have been implicated in adhesion to and penetration of host cells and in modulation of the immune system of the mammalian host (Travassos & Almeida 1993, Almeida & Gazzinelli 2001, Previato et al. 2004, Buscaglia et al. 2006). Among the most abundant glycoproteins on the T. cruzi cell surface are Tc-mucins, trans-sialidase (TS) family members and the mucin-associated surface proteins (MASPs) (Bartholomeu et al. 2009,). It is estimated that there are about 850 mucin-like genes, which, when grouped with the TSs, TS-like genes and MASPs, correspond to about 7% of the T. cruzi genome (El-Sayed et al. 2005).

The Tc-mucins are highly O-glycosylated, threonine-rich glycoproteins (Almeida et al. 1994, Previato et al. 1994, 1995, Acosta-Serrano et al. 1995, Di Noia et al. 1996, Mendonça-Previato et al. 2008) that are glycosylphosphatidylinositol-anchored to the parasite surface (Previato et al. 1995). O-linked oligosaccharides account for up to 60% of the mucin molecular mass (Previato et al. 1994) and are linked to Thr residues through an N-acetylglucosamine (?-GlcNAc) unit, which is added by an unusual ?-O-N-acetylglucosaminosyl transferase (Previato et al. 1998). The parasite mucins incorporate ?2,3-linked sialic acid from exogenous sialyl-glycoconjugates (Previato et al. 1985, 1990) by means of a glycoside hydrolase TS (Previato et al. 1985). Sialylation of the T. cruzi surface by the enzyme TS is pivotal for the establishment of infection (Frasch 2000).

Up to the present time, the O-glycan chains of T. cruzi mucins have been characterised from the epimastigote of five strains. O-glycan chains are composed of GlcNAc, galactose (Gal) and sialic acid (Neu5Ac); however, they can vary in complexity and in the ring form of the galactosyl residue in a strain-dependent manner (Previato et al. 1994, 1995, Todeschini et al. 2001, Agrellos et al. 2003, Jones et al. 2004). It has been verified that whilst O-glycans from the mucins of Y (Previato et al. 1995) and CL Brener (Todeschini et al. 2001) strains do not contain ?-Galf residues as a substituent on the ?-GlcNAc O-4, this substituent is observed in G, Dm28c and Tulahuen strains (Previato et al. 1994, Agrellos et al. 2003, Jones et al. 2004). Several laboratories have reported the division of T. cruzi into two major groups with distinct designations. To homogenise the nomenclature, the groups were named T. cruzi I and T. cruzi II by consensus (Anonymous 1999). Interestingly, Tc-mucin O-glycan core structures correlate with the classification of T. cruzi strains based on the size of the PCR products of the 24S alpha rRNA and the mini-exon genes (Souto et al. 1996). Strains whose mucins contain Galf (G, Dm28c and Tulahuen) group together in T. cruzi I and the strains lacking Galf are classified as T. cruzi II (Jones et al. 2004).

The Colombiana strain, isolated from a chronic human case in Colombia (Federici et al. 1964), is particularly interesting since it is resistant to drugs used to treat Chagas disease (Murta et al. 1998). Genetic analysis based on the dimorphism of the 24S alpha rRNA gene and the intergenic region of the mini-exon gene (Souto et al. 1996) indicated that the Colombiana strain belongs to the T. cruzi I group. This work reports a structural study of the mucins from epimastigote forms of T. cruzi Colombiana strain and emphasises the potential use of structural data as a taxonomic parameter.



T. cruzi growth - Epimastigote forms of T. cruzi Colombiana strain were cultivated in brain heart infusion medium supplemented with 10 mg/L hemin, 20 mg/L folic acid and 5% (v/v) foetal calf serum at 28ºC under constant agitation. Cultures were harvested during the exponential phase of growth and frozen as previously described (Previato et al. 1994).

Isolation and purification of Tc-mucin - Frozen cells (50 g wet weight) were thawed and extracted twice with cold water (150 mL) and the pellet was recovered by centrifugation (12,000 x g, 10 min, 4°C). The pellet was extracted with 45% (v/v) aqueous phenol at 75°C for 15 min. The aqueous phase was dialysed against tap water, lyophilised, dissolved in water and applied to a 4.7 x 25 cm column of Bio-Gel P-60. Excluded material containing carbohydrate was freeze-dried and the mucin was recovered in the insoluble phase after extraction (twice) with chloroform/methanol/water (10:10:3, v/v/v) (Previato et al. 1994). Water-soluble material in the chloroform/methanol/water insoluble components was recovered and lyophilised. The homogeneity of the mucin was determined by electrophoresis in a 15% polyacrylamide gel in the presence of sodium dodecyl sulphate (SDS-PAGE) (Laemmli 1970) and the bands were visualised by staining with periodate-Schiff (PAS) reagent (Fairbanks et al. 1971).

Isolation and fractionation of O-glycan-alditols -Tc-mucin was submitted to base-catalysed ?-elimination with concomitant reduction using 20 mM NaOH and 0.3 M NaBH4 at 37°C for 24 h. The solution was then neutralised with 50% (v/v) acetic acid, passed through a Dowex 50W-X8 ion-exchange column and lyophilised (Previato et al. 1994). The material was dissolved in water and fractionated on a Bio-Gel P-4 column (1 x 110 cm). Fractions of 1.25 mL were collected and the carbohydrate elution was monitored by spotting 4-?L portions onto a thin layer chromatography plate and staining with orcinol-H2SO4 spray (Humbel & Collart 1975). Carbohydrate-containing fractions were further fractionated by high performance liquid chromatography on porous graphitic carbon as previously described (Jones et al. 2004).

Carbohydrate analysis - The neutral sugar composition was determined after methanolysis in 0.5 M methanolic HCl as previously described (Previato et al. 1994).

Amino acid analysis - Amino acid analysis was determined after acid hydrolysis in 5.6 N HCl at 110ºC for 22 h in an autoanalyser by the method of Fauconnet and Rochemont (1978).

Nuclear magnetic resonance spectroscopic (NMR) analysis - Tc-mucin O-glycan samples were analysed by NMR on a Varian Unity 500 spectrometer as previously described (Previato et al. 1994).



Isolation of Tc-mucin, amino acid and carbohydrate analysis - Mucin from epimastigote forms of T. cruzi Colombiana strain was extracted with aqueous phenol and recovered from the water-soluble fraction of the chloroform/methanol/water insoluble components. Analysis of this material by SDS-PAGE revealed diffuse PAS staining of bands with apparent molecular masses of 45 and 43 kDa.

Carbohydrate analysis of the Tc-mucins indicated the presence of Gal, GlcNAc and mannose (Man) in a molar ratio of 4.0:1.3:1.0 along with Neu5Ac. Amino acid analysis demonstrated Thr to be the predominant amino acid, contributing 42% of the total, whilst Asp, Glu, Ala, Pro contributed 12.5%, 12.6%, 8.8%, 8.1% and 7.2%, respectively. All other remaining amino acids comprised 8.3% of the total.

O-linked glycans from Tc-mucins - O-glycans were released from mucins by reductive ?-elimination and fractionated on a BioGel P4 column. The resulting six carbohydrate-containing fractions were analysed by NMR spectroscopy. The O-glycan alditols were identified by comparing their one-dimensional 1H spectra (Fig. 1) with those of previously characterised O-glycans (Mendonça-Previato et al. 2008). All of the fractions also contained a ManNAc-ol analogue, a product of the base-catalysed epimerisation of GlcNAc (Jones et al. 2000), as was evident from a chemical shift of the N-acetyl methyl resonance at 2.03 ppm for ManNAc-ol and 2.05 ppm for the GlcNAc epimers (data not shown).



The lowest molecular mass material was identified as GlcNAc-ol, whilst the monosaccharide alditol fraction contained a single major component identified as Galf?1?4GlcNAc-ol (data not shown). This was evidenced by a single intense anomeric signal at a lowfield position (5.154 ppm) with a small coupling constant (3JH1,H2 = 2.3 Hz) characteristic of a ?-Galf residue. This compound was previously characterised in the reductive cleavage products of the mucins from T. cruzi G-strain (Acosta-Serrano et al. 1995), the Dm28c clone (Agrellos et al. 2003) and the Tulahuen strain (Jones et al. 2004).

The disaccharide alditol fraction contained two components present in an approximate ratio of 6:4 (Fig. 1A). The minor component was Galp?1?2Galf?1?4ManNAc-ol, which had been observed in a study of the Tulahuen strain (Jones et al. 2004). The major component contains the presence of an anomeric signal at 4.45 ppm, typical of a terminal ?-Galp residue. Disubstitution of the GlcNAc-ol at O-4 and O-6 was determined by comparison of the 1D 1H NMR spectrum with that of the disaccharide alditol previously isolated from G (Previato et al. 1994) and Tulahuen strain mucins (Jones et al. 2004) and is consistent with the Galf?1?4 [Galp?1?6]GlcNAc-ol structure.

Compared with the disaccharide alditol, the one-dimensional 1H NMR spectrum of the trisaccharide alditol fraction (Fig. 1B) contained an additional anomeric signal at 4.62 ppm, characteristic of a terminal Galp?1?3 residue, and a shift of the Galp?1?6 H-1 resonance from 4.45 ppm to 4.50 ppm. This component was therefore identified as the known compound Galf-?-(1?4)[Galp-?(1?3)-Galp-?-(1?6)]GlcNAc-ol (Previato et al. 1994, Jones et al. 2004).

The 1D 1H spectra of the tetrasaccharide alditol fraction (Fig. 1C) showed the presence of a single ?-Galf spin system and three ?-Galp spin systems at chemical shifts identical to those previously reported for the structure of Galf?1?4{Galp?1?3[Galp ?1?2]Galp?1?6}GlcNAc-ol in studies of the O-glycans from T. cruzi G (Previato et al. 1994) and Tulahuen strain mucins (Jones et al. 2004).

The most complex structure isolated was a pentasaccharide alditol. The 1H NMR spectra of this fraction (Fig. 1D) showed an additional ?-Galp spin system and a downfield shift of the ?-Galf H-1 and H-2 from 5.3 - 5.5 ppm (?-Galf H-1) and from 4.08 - 4.41 ppm (H-2). These data, and the TOCSY spectrum (datum not shown), are identical with those previously reported for the structure of Galp?1?2Galf?1?4 {Galp?1?3[Galp?1?2]Galp ?1?6}GlcNAc-ol from the G-strain (Previato et al. 1994).



T. cruzi infects a broad range of hosts including wild and domestic animals. In humans, it causes Chagas disease and has a variety of clinical presentations. The severity and symptoms of the disease vary with the geographical region and are related to the characteristics of the patient and the parasite. T. cruzi strains diverge in many other aspects, including tissue tropism, morphology, infectivity, isoenzymes and drug resistance. Early studies provided evidence that T. cruzi is diploid, that sexual reproduction is rare or absent and that the population evolved from independent clonal lineages (Tibayrenc et al. 1986, Tibayrenc & Ayala 1988). Indeed, the remarkable genetic diversity observed raises the question of whether all of these strains are part of a single species or include more than one (Macedo & Pena 1998). Together, these observations require a means to group the strains and a number of classifications have resulted. Molecular typing of T. cruzi strains based on the size of the PCR products of the 24S alpha rRNA and the mini-exon genes (Souto et al. 1996) indicated that the G, Dm28c and Tulahuen strains belong to the T. cruzi I Group and that the Y and CL Brener strains belong to the T. cruzi II Group. Nevertheless, it should be emphasised that CL Brener, the reference strain of the T. cruzi genome project, is a hybrid (El Sayed et al. 2005).

Great efforts have been made to understand this heterogeneity. Biological and epidemiological studies provide support for an association of T. cruzi Group II with the domestic cycle, which is mostly involved in human infections, while T. cruzi Group I is associated with the sylvatic cycle, which is asymptomatic in humans most of the time (Briones et al. 1999). The observed variability during infection by different T. cruzi strains is most probably a result of the diversity in parasite/host interactions resulting from variability of the molecules expressed on both the parasite and host cell surfaces. Mucins are among the most abundant glycoproteins on the T. cruzi surface. They are Thr-rich, highly O-glycosylated glycoproteins. The Colombiana strain mucins appear as a double band of 43/45 kDa by SDS-PAGE. The O-glycan chains (Fig. 2) coincide with the Tc-mucins described on strains classified as belonging to T. cruzi group I (Anonymous 1999) (Tulahuen, Dm28c, G and Tulahuen) and contain Galf?1?4 substituents (Previato et al. 1994, Agrellos et al. 2003, Jones et al. 2004), which have not been observed in T. cruzi group II (Anonymous 1999) strains (Y and CL Brener) (Previato et al. 1995, Todeschini et al. 2001). These observations may be of epidemiologic significance since mammalian hosts do not produce glycoconjugates containing Galf and present a strong immune response against this epitope (Mendonça-Previato et al. 1983, Golgher et al. 1993).

Together, these data suggest that structural biology data can be used as an approach to classify/identify T. cruzi strains. Herein we emphasise the importance of accumulating structural data for such comparisons and propose the use of these data as a taxonomic parameter in the differentiation of the T. cruzi clones (Fig. 3).




Almeida IC, Ferguson MAJ, Schenkman S, Travassos LR 1994. Lytic anti-alpha-galactosyl antibodies from patients with chronic Chagas´disease recognize novel O-linked oligosaccharides on mucin-like glycosylphosphatidylinositol anchored glycoprotein of Trypanosoma cruzi. Biochem J 304: 793-803.

Almeida IC, Gazzinelli RT 2001. Proinflammatory activity of glycosylphosphatidylinositol anchors derived from Trypanosoma cruzi: structural and functional analyses. J Leuk Biol 70: 467-477.

Anonymous 1999. Recommendations from a satellite meeting. Mem Inst Oswaldo Cruz 94 (Suppl. 1): 429-432.

Acosta-Serrano A, Schenkman S, Yoshida N, Mehlert A, Richardson JM, Fererguson MAJ 1995. The lipid structure of the glycophosphatidylinositol-anchored mucin-like sialic acid acceptors of Trypanosoma cruzi changes during parasite differentiation from epimastigotes to infective metacyclic trypomastigote forms. J Biol Chem 270: 27244-27253.

Agrellos OA, Jones C, Previato JO, Mendonça-Previato L 2003. A novel sialylated and galactofuranose-containing O-linked glycan, Neu5Ac?2?Galp?1?6(Galf?1?4)GlcNAc, is expressed on the sialoglycoprotein of Trypanosoma cruzi Dm28c. Mol Biochem Parasitol 126: 93-96.

Bartholomeu DC, Cerqueira GC, Leão AC, Daroch WD, Pais FS, Macedo C. Djikeng A, Teixeira SM, El-Sayed NM 2009. Genomic organization and expression profile of the mucin-associated surface protein (MASP) family of the pathogen Trypanosoma cruzi. Nucleic Acids Res 37: 3407-3417.

Briones MR, Souto RP, Stolf BS, Zingales B 1999. The evolution of two Trypanosoma cruzi subgroups inferred from rRNA genes can be correlated with the interchange of American mammalian faunas in the Cenozoic and has implication to pathogenicity and host specificity. Mol Biochem Parasitol 104: 219-232.

Buscaglia CA, Campo VA, Frasch AC, Di Noia JM 2006. Trypanosoma cruzi surface mucins: host-dependent coat diversity. Nat Rev Microbiol 4: 229-236.

Di Noia JM, Pollevick GD, Xavier MT, Previato JO, Mendonça-Previato L, Sanchez DO, frasch AC 1996. High diversity in mucin genes and mucin molecules in Trypanosoma cruzi. J Biol Chem 271: 32078-32083.

El-Sayed NM, Myler PJ, Bartholomeu DC, Nilsson D, Aggarwal G, Tran AN, Ghedin E, Worthey EA, Delcher AL, Blandin G, Westenberger SJ, Caler E, Cerqueira GC, Branche C, Haas B, Anupama A, Arner E, Aslund L, Attipoe P, Bontempi E, Bringaud F, Burton P, Cadag E, Campbell DA, Carrington M, Crabtree J, Darban H, da Silveira JF, de Jong P, Edwards K, Englund PT, Fazelina G, Feldblyum T, Ferella M, Frasch AC, Gull K, Horn D, Hou L, Huang Y, Kindlund E, Klingbeil M, Kluge S, Koo H, Lacerda D, Levin MJ, Lorenzi H, Louie T, Machado CR, McCulloch R, McKenna A, Mizuno Y, Mottram JC, Nelson S, Ochaya S, Osoegawa K, Pai G, Parsons M, Pentony M, Pettersson U, Pop M, Ramirez JL, Rinta J, Robertson L, Salzberg SL, Sanchez DO, Seyler A, Sharma R, Shetty J, Simpson AJ, Sisk E, Tammi MT, Tarleton R, Teixeira S, Van Aken S, Vogt C, Ward PN, Wickstead B, Wortman J, White O, Fraser CM, Stuart KD, Andersson B 2005. The genome sequence of Trypanosoma cruzi etiologic agent of Chagas´ disease. Science 309: 409-415.

Fairbanks G, Steck TL, Wallach DFH 1971. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry 10: 2606-2616.

Fauconnet M, Rochemont J 1978. A single-column amino acid analysis method which resolves hexosamines and several cysteine derivatives. Anal Biochem 91: 403-409.

Federici EE, Abelmann WH, Neva FA 1964. Chronic and progressive myocarditis in C3H mice infected with Trypanosoma cruzi. Am J Trop Med Hyg 13: 272-280.

Frasch ACC 2000. Functional diversity in the trans-sialidase and mucin families in Trypanosoma cruzi. Parasitol Today 16: 282-286.

Golgher DB, Colli W, Souto-Padrón T, Zingales B 1993. Galactofuranose-containing glycoconjugates of epimastigote and trypomastigote forms of Trypanosoma cruzi. Mol Biochem Parasitol 60: 249-264.

Hotez PJ 2008. Neglected infections of poverty in the United States of America. PLoS Negl Trop Dis 2: e256.

Humbel R, Collart M 1975. Oligosaccharides in urine of patients with glycoprotein storage diseases. Rapid detection by thin-layer chromatography. Clin Chem Acta 60: 143-145.

Jones C, Previato JO, Mendonça-Previato L 2000. NMR assignments for glucosylated and galactosylated N-acetylhexosaminitols: oligosaccharide alditols related to O-linked glycans from the protozoan parasite Trypanosoma cruzi. Carbohydr Res 328: 321-330.

Jones C, Todeschini AR, Agrellos OA, Previato JO, Mendonça-Previato L 2004. Heterogeneity in the biosynthesis of mucin O-glycans from Trypanosoma cruzi Tulahuen strain with the expression of novel galactofuranosyl-containing oligosaccharides. Biochemistry 43: 11889-11897.

Laemmli UK 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685.

Macedo AM, Pena SDJ 1998. Genetic variability of Trypanosoma cruzi: implications for the pathogenesis of Chagas´ Disease. Parasitol Today 14: 119-124.

Mendonça-Previato L, Gorin PAJ, Braga AF, Scharfstein J, Previato JO 1983. Chemical structure and antigenic aspects of complexes obtained from epimastigotes of Trypanosoma cruzi. Biochemistry 22: 4980-4987.

Mendonça-Previato L, Todeschini AR, Heise N, Agrellos AO, Dias WB, Previato JO 2008. Chemical structure of major glycoconjugates from parasites. Curr Org Chem 12: 926-939.

Murta SMF, Gazzinelli RT, Brener Z, Romanha AJ 1998. Molecular characterization of susceptible and naturally resistent strains of Trypanosoma cruzi to benznidazole and nifurtimox. Mol Biochem Parasitol 93: 203-214.

Previato JO, Andrade AF, Pessolani MC, Mendonça-Previato L 1985. Incorporation of sialic acid into Trypanosoma cruzi macromolecules. A proposal for a new metabolic route. Mol Biochem Parasitol 16: 85-96.

Previato JO, Andrade AFB, Vermelho A, Firmino JC, Mendonça-Previato L 1990. Evidence for N-glycolylneuraminic acid incorporation by Trypanosoma cruzi from infected animal. Mem Inst Oswaldo Cruz 85: 38-39.

Previato JO, Jones C, Gonçalves LPB, Wait R, Travassos LR, Mendonça-Previato L 1994. O-glycosidically linked N-acetylglucosamine-bound oligosaccharides from glycoproteins of Trypanosoma cruzi. Biochem J 301: 151-159.

Previato JO, Jones C, Xavier MT, Wait R, Travassos LR, Parodi AJ, Mendonça-Previato L 1995. Structural characterization of the major glycosylphosphatidylinositol membrane-anchored glycoprotein from epimastigote forms of Trypanosoma cruzi Y-strain. J Biol Chem 270: 7241-7250.

Previato JO, Sola-Penna M, Agrellos OA, Jones C, Oeltmann T, Travassos LR, Mendonça-Previato L 1998. Biosynthesis of O-N-Acetylglucosamine-linked glycans in Trypanosoma cruzi. J Biol Chem 273: 14982-14988.

Previato JO, Wait R, Jones C, DosReis GA, Todeschini AR, Heise N, Mendonça-Previato L 2004. Glycoinositolphospholipid from Trypanosoma cruzi: structure, biosynthesis and immunobiology. Adv Parasitol 56: 1-41.

Riera C, Guarro A, Kassab HE, Jorba JM, Castro M, Angrill R, Gállego M, Fisa R, Martin C, Lobato A, Portús M 2006. Congenital transmission of Trypanosoma cruzi in Europe (Spain): a case report. Am J Trop Med Hyg 75: 1078-1081.

Souto RP, Fernandes O, Macedo AM, Campbell DA, Zingales B 1996. DNA markers define two major phylogenetic lineages of Trypanosoma cruzi. Mol Biochem Parasitol 83: 141-152.

Tibayrenc M, Ayala FJ 1988. Isoenzyme variability in Trypanosoma cruzi, the agent of Chagas' disease: genetic, taxonomical and epidemiological significance. Evolution 42: 277-292.

Tibayrenc M, Ward P, Moya A, Ayala FJ. 1986. Natural populations of Trypanosoma cruzi, the agent of Chagas´ disease, have a complex multiclonal structure. Proc Natl Acad Sci USA 83: 293-296.

Todeschini AR, Jones C, Wait R, Previato JO, Mendonça-Previato L 2001. Structures of O-glycosidically linked oligosaccharides from glycoproteins of Trypanosoma cruzi CL Brener strain: evidence for the presence of O-linked sialyloligosaccharides. Glycobiology 11: 47-55.

Travassos LR, Almeida IC 1993. Carbohydrate immunity in American trypanosomiasis. Spring Sem Immunopathol 15: 183-204.

WHO - World Health Organization 2007. Reporte Del grupo de trabajo cientifico sobre la enfermedad de Chagas (Acessed July 2007). Avaliable from:

Received 12 May 2009
Accepted 16 June 2009
Financial support: CNPq, FAPERJ, National Institute for Science and Technology in Vaccines ART and EGA contributed equally to this study.


+ Corresponding author:

Our Location

Memórias do Instituto Oswaldo Cruz

Av. Brasil 4365, Castelo Mourisco 
sala 201, Manguinhos, 21040-900 
Rio de Janeiro, RJ, Brazil

Tel.: +55-21-2562-1222

This email address is being protected from spambots. You need JavaScript enabled to view it.

Support Program


fiocruz governo
faperj cnpq capes