Figure 1.
A) Experimental timeline B) Core temperature measurements prior to and post the 8th rhGAA IV injection in wt 129 SVE and wt BALB/c mice (n = 5) C) Anti-rhGAA IgG1 antibody in 129SVE wt mice D) Anti-rhGAA IgG1 antibody in BALB/c mice E) Anti-rhGAA IgG2a antibody in wt 129SVE mice F) Anti-rhGAA IgG2a antibody in wt BALB/c mice.
Figure 2.
Vital signs measured by pulse oxymetry, prior to and 5 minutes after the 8th rhGAA IV injection in wt BALB/c and wt 129SVJ mice (n = 5).
A) Percentage oxygen saturation B) Heart rate C) Breath distention D) Pulse distention, p<0.05 *, ns = not significant, (inj; injection).
Figure 3.
Antibody responses to varying doses of rhGAA in null mutation (n = 6) or P545L mutant mice (n = 5).
A) Anti-rhGAA IgG1 in 1 mg/kg rhGAA injected GAA-/- 129SVE mice tested weekly B) Anti-rhGAA IgG1 in 5 mg/kg rhGAA injected GAA-/- 129SVE mice C) Anti-rhGAA IgG1 in 20 mg/kg rhGAA injected GAA-/- 129SVE mice D) Anti-rhGAA IgG1 response in 20 mg/kg rhGAA injected P545L mice E) Anti-rhGAA IgG2a in 1 mg/kg rhGAA injected GAA-/- 129SVE mice F) Anti-rhGAA IgG2a in 5 mg/kg rhGAA injected GAA-/- 129SVE mice G) Anti-rhGAA IgG2a in 20 mg/kg rhGAA injected GAA-/- 129SVE mice H) Anti-rhGAA IgG2a in 20 mg/kg rhGAA injected P545L mice. Arrows indicate fold decrease over corresponding 20 mg/kg cohort time point. p<0.05 *, p<0.005 **, p<0.0005 ***, ns = not significant.
Figure 4.
Temperature measurements prior to and post rhGAA IV injections in A-C) GAA-/-129SVE mice (n = 6) receiving 1 mg/kg, 5 mg/kg or 20 mg/kg doses of rhGAA IV D) GAA-/-129SVE mice injected with PBS E) P545L mice (n = 5) receiving 20 mg/kg of rhGAA IV, F) Experimental timeline indicating GAA-/- 129SVE mice or P545L C57BL/6 x 129SVE mice injected with 1 mg/kg, 5 mg/kg or 20 mg/kg doses of rhGAA G) Survival curve.
Figure 5.
Comparison of pulse oxymetry measurements of vital signs prior to and post the 4th rhGAA (1 mg/kg, 5 mg/kg or 20 mg/kg) ERT IV injection in GAA-/- 129SVE (n = 6) and P545L mice (20 mg/kg rhGAA; n = 5).
A) oxygen saturation B) heart rate C) pulse distention D) breath distention, E) Time taken for the formation of a platelet plug post-injury (tail-snip) prior to and post-rhGAA IV administration. Changes in hemorheologic values from complete blood counts in 129SVE GAA-/- mice and P545L naive and 20 mg/kg rhGAA injected mice (5 min post rhGAA injection). F) Hematocrit, G) Hemoglobin, p<0.05 *, p<0.005 **, p<0.0005 ***, ns = not significant.
Figure 6.
A) Reduction of clotting times measured in activated partial thromboplastin time (aPTT) in GAA-/- 129SVE and P545L mice. B) D-Dimer levels in GAA-/-129SVE naïve and rhGAA injected mice. Changes in hemorheologic values from complete blood counts in 129SVE GAA-/- mice and P545L naive and 20 mg/kg rhGAA injected mice (5 min post rhGAA injection). C) Platelet count, D) Mean platelet volume, E) Platelet width distribution, p<0.05 *, p<0.005 **, p<0.0005 ***, ns = not significant.
Figure 7.
Representative examples of hematoxylin and eosin (H&E) staining of paraffin embedded sections of liver, kidney and heart (n = 3) in A-C) naïve GAA-/- 129SVE mice and D-F) 20 mg/kg rhGAA IV injected GAA-/- 129SVE indicating residual RBC, G-I) 20 mg/kg rhGAA IV injected P545L mice.
Figure 8.
Representative examples of fibrinogen staining in paraffin embedded sections of liver, kidney and heart (n = 3) in A-C) naïve GAA-/-129SVE mice and D-F) 20 mg/kg rhGAA IV injected GAA-/- 129SVE and G-I) 20 mg/kg rhGAA IV injected P545L mice.