Peptide-based analysis of the amino acid sequence important to the immunoregulatory function of Trypanosoma cruzi Tc52 virulence factor.
The intracellular protozoan parasite Trypanosoma cruzi is the aetiological agent of Chagas' disease. We have previously identified a T. cruzi-released protein called Tc52, which is crucial for parasite survival and virulence. In the present study, we attempted to define the Tc52 epitope(s) responsible for its immunoregulatory function. A naturally occurring major peptide fragment of molecular mass 28 kDa (Tc28k) was identified, which was localized in the C-terminal portion of Tc52 and was inhibitory for T-cell activation. Synthetic peptides corresponding to amino acid sequences in Tc52 were evaluated for their ability to modulate T-cell proliferation and cytokine production. Results obtained using five peptides spanning the N-terminal or C-terminal domain of the Tc52 protein indicated that the activity mapped to Tc52 residues 432-445. Moreover, it was found that the peptide, when coupled to a carrier protein (ovalbumin), exhibited increased inhibitory activity on T-lymphocyte activation. Incubation with 8 nm ovalbumin-coupled peptide 432-445 resulted in approximately the same levels (>75%) of inhibition of T-cell proliferation as 5 micro g/ml Tc28k. Furthermore, we showed that the coupled peptide significantly down-regulated the secretion of interferon-gamma (IFN-gamma) and interleukin-2 (IL-2). Likewise, in immunized mice, the coupled peptide 432-445 was a very poor B- and T-cell antigen compared with the other Tc52-derived peptides. These results suggest that the immunomodulatory portion of the T. cruzi Tc52 virulent factor may reside, at least in part, in a conserved sequence within its C-terminal domain, which could minimize its antigenicity.
Borges
Margarida
M
IRD UR 008 Pathogénie des Trypanosomatidae, Centre IRD de Montpellier, France.
Da Silva
Anabela Cordeiro
AC
Sereno
Denis
D
Ouaissi
Ali
A
D000595:Amino Acid Sequence; D000818:Animals; D002455:Cell Division / Q000276:immunology; D016207:Cytokines / Q000096:biosynthesis; D015536:Down-Regulation; D007108:Immune Tolerance; D007371:Interferon-gamma / Q000096:biosynthesis; D007376:Interleukin-2 / Q000096:biosynthesis; D008198:Lymph Nodes / Q000276:immunology; D008213:Lymphocyte Activation; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008808:Mice, Inbred CBA; D010047:Ovalbumin / Q000276:immunology; D010446:Peptide Fragments / Q000276:immunology; D015800:Protozoan Proteins / Q000737:chemistry / Q000276:immunology*; D013154:Spleen / Q000276:immunology; D014349:Trypanosoma cruzi / Q000276:immunology*; D014774:Virulence / Q000276:immunology
C097550:52-kDa protein, Trypanosoma cruzi; D016207:Cytokines; D007376:Interleukin-2; D010446:Peptide Fragments; D015800:Protozoan Proteins; D007371:Interferon-gamma; D010047:Ovalbumin