Memórias do Instituto Oswaldo Cruz · 1998 · 27 citations · 27 references
BiologyTrypanosoma CruziParasitic DiseaseMetacyclic TrypomastigotesLong TimeAfrican TrypanosomiasisParasitic ProtozoaPathogenesisImmunologyPathologyMolecular GeneticsAxenic CultureMedicineCell BiologyCulture Modify GeneParasitologyHost-parasite Relationship
In this study we examined whether the maintenance of Trypanosoma cruzi by long-time in axenic culture produces changes in gene expression and antigenic profiles. The studies were made with a Dm30L-clone from a low-virulent strain and a non-cloned virulent EP-strain of T. cruzi. Both parasites were maintained, for at least seven years, by successive alternate passage triatomine/mouse (triatomine condition), or by serial passage in axenic medium (culture condition). The comparison of the [35S]methionine metabolic labeling products of virulent and non-virulent parasites by 2D-SDS-PAGE, clearly indicates that the expression of metacyclic trypomastigotes (but not of epimastigotes) proteins have been altered by laboratory maintenance conditions. Western blot analysis of EP and Dm30L-epimastigotes using a serum anti-epimastigotes revealed that although most of antigens are conserved, four antigens are characteristics of triatomine condition parasites and three other are characteristics of culture condition parasites. Anti-metacyclics serum revealed significative differences in EP- and Dm30L-metacyclic trypomastigotes from triatomine condition. However, avirulent metacyclic forms were antigenically very similar. These results suggest that besides a possible selection of avirulent subpopulation from T. cruzi strains genetically heterogeneous when maintained by long time in axenic culture, changes in virulence might be due to post-translational modifications of the antigens induced by the absence of the natural alternability (vertebrate-invertebrate) in the life-cycle of T. cruzi.
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Harry Towbin, T. Staehelin, J. Gordon · Proceedings of the National Academy of Sciences · 1979 · 53.8K citations · Full text
Immunocytochemical Technique, Glycobiology, Polyacrylamide Gels +19
High resolution two-dimensional electrophoresis of proteins.
Patrick H. O’Farrell · Journal of Biological Chemistry · 1975 · 19.3K citations · Full text