Mouse Anti-TcP2β Recombinant Antibody (clone mAb 17.2)

A. Representative histogram showing the HEK and HEK-β1 cells labelled with mAb 17.2 followed by a Cy3-conjugated goat anti-mouse IgG. B. Results are expressed as means ± SD (n = 3). Inset: Binding of mAb 17.2 to HEK-β1 cells in the presence of R13 peptide. ** p<0.01; *** p<0.001. C. Passive transfer of mAb 17.2 to naïve mice. Repolarisation abnormalities (a) and first degree AV conduction block (b) are indicated by arrows. bpm: beats per minute.

Pizarro, J. C., Boulot, G., Bentley, G. A., Gomez, K. A., Hoebeke, J., Hontebeyrie, M., ... & Smulski, C. R. (2011). Crystal structure of the complex mAb 17.2 and the C-terminal region of Trypanosoma cruzi P2β protein: implications in cross-reactivity. PLoS neglected tropical diseases, 5(11), e1375.

A. Sequence aligment of T. cruzi R13 and P015 peptides with the mammalian counterpart H13 peptide. White letters correspond to residues necessary for antibody recognition, as identified by alanine scanning. Grey background corresponds to those residues conserved in the other two peptides. B. Inhibition of the interaction between scFv C5 and TcP2β protein by R13, P015 and H13 peptides, using surface plasmon analysis. The figure corresponds to one representative result out of 3 independent assays. C. Crystal structure of the complex mAb 17.2-R13 (PDB 3SGE). D. Model of the interaction of mAb 17.2 with the H13 peptide. The interaction energy is indicated on each case. E. NMR solution structure of H13 peptide (PDB 1S4J) in comparison with the antibody-bound peptide structure modeled.

Ayub, M. J., Nyambega, B., Simonetti, L., Duffy, T., Longhi, S. A., Gómez, K. A., ... & Smulski, C. R. (2012). Selective Blockade of Trypanosomatid Protein Synthesis by a Recombinant Antibody Anti-Trypanosoma cruzi P2β Protein. PloS one, 7(5), e36233.

Enzymatic activity was performed on 1.5 μg of ROS membranes preparation (0.5 μM rhodopsin) in a final volume of 100 μL. A) IgG fractions from control patients scarcely inhibited cGMP-PDE activity (lanes 1–5). No inhibition of rhodopsin-dependent cGMP-PDE activity was produced by IgG reactive to M2 (lanes 6–7). IgG fractions from cChHD patients reactive to β1-AR significantly reduced the enzymatic activity (lanes 8–15), which was restored when preincubated with T. cruzi total antigen (lanes16–23). Reversion of inhibitory effect is also demonstrated with R13 (lanes 24, 25) and H26R (lanes 26, 27) peptides. No effect on enzyme activity light stimulation is produced by T. cruzi lysate (lane 28). B) Light-induced cGMP-PDE activity is strongly inhibited when preincubated with mAb E2. The magnitude of inhibition caused by mAb M16 was similar to that produced by the specific mAb E2. This inhibition was abolished when mAb E2 and M16 were preincubated with E2 and H26R peptides. Monoclonal antibody 17.2 showed similar inhibitor effect, which diminished by preincubation with R13 peptide. No inhibition was observed with mAb M2. Inset) Rhodopsin-dependent cGMP-PDE activity is expressed as the difference between light-stimulated (▪) activity and basal activity (□) recorded under continue slow intensity red light (dark condition). Results shown here represent the mean of at least 3 experiments. Standard deviation was calculated with GraphPad Prism software, San Diego.

Matsumoto, S. C., Labovsky, V., Roncoroni, M., Guida, M. C., Giménez, L., Mitelman, J., ... & Levin, M. J. (2006). Retinal dysfunction in patients with chronic Chagas’ disease is associated to anti-Trypanosoma cruzi antibodies that cross-react with rhodopsin. The FASEB journal, 20(3), 550-552.

Apoptosis was measured by TUNEL assay. DAPI staining displays all of the nuclei in the field (400). Ten random fields were analysed and the percentage of TUNEL-positive cells was calculated as indicated in Section 2.10.3. Results are means ± S.E.M. (n = 3).

Levy, G. V., Tasso, L. M., Longhi, S. A., Rivello, H. G., Kytö, V., Saukko, P., ... & Gómez, K. A. (2011). Antibodies against the Trypanosoma cruzi ribosomal P proteins induce apoptosis in HL-1 cardiac cells. International journal for parasitology, 41(6), 635-644.

The mRNA expression levels of the pro-apoptotic molecule (Bax) and anti-apoptotic molecule (BclXL) were normalised to glyceraldehyde 3-phosphate dehydrogenase (GAPDH). Results are mean ± S.E.M. (n = 3).

Levy, G. V., Tasso, L. M., Longhi, S. A., Rivello, H. G., Kytö, V., Saukko, P., ... & Gómez, K. A. (2011). Antibodies against the Trypanosoma cruzi ribosomal P proteins induce apoptosis in HL-1 cardiac cells. International journal for parasitology, 41(6), 635-644.