Recombinant Enolase of Trypanosoma cruzi as a Novel Vaccine Candidate against Chagas Disease in a Mouse Model of Acute Infection

Figure 1

T. cruzi enolase immunodetection. (a) Total extract of T. cruzi soluble proteins and its respective replica for western blotting. Lane M: molecular weight marker; lanes 1 and 3: total soluble proteins (10 μg); lanes 2 and 4: total soluble proteins (20 μg); lanes 5 and 6: western blot of immobilized proteins with polyclonal antibodies anti-rTcENO; lanes 7 and 8: western blot of immobilized proteins with polyclonal antibodies anti-pBKTcENO. (b) Purified rTcENO and its respective replica for western blotting. Lane M: molecular weight marker; lanes 1 and 2: rTcENO (10 μg); lane 3: western blot with pool of polyclonal antibodies anti-rTcENO; lane 4: western blot with pool of polyclonal antibodies anti-pBKTcENO. The arrowhead indicates the signal for a band of approximately 46 kDa (the TcENO estimated weight). (c) Indirect immunofluorescence assay. The secondary antibody that recognized the anti-rTcEno was FITC labeled (green fluorescence). Nuclear and kinetoplast DNA were stained with DAPI and shown by blue fluorescence. Each image is a representative of at least two independent experiments and captured by confocal microscope. (A) Nonpermeabilized parasite with anti-T. cruzi polyclonal antibodies; (B) nonpermeabilized parasite with anti-rTcENO polyclonal antibodies; (C) permeabilized parasite with anti-T. cruzi polyclonal antibodies; (D) permeabilized parasite with anti-rTcENO polyclonal antibodies.

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