Figure 1.
An overlay of sensograms showing Polymer A interaction with gliadin at a series of concentrations of 10-640 ng/mL.
KD was determined to be 12.1 nM, with kon of 2.39×105 1/M·s and koff of 2.89×10-3 1/s. B. An overlay of sensograms showing Polymer B interacting with gliadin, with KD of 2.44 nM, kon of 6.11×105 1/M·s and koff of 1.49×10-3 1/s using Biacore Evaluation Software (see Methods). For both graphs, colored lines indicate experimental sensograms at various concentrations, while corresponding black lines denote fitting of experimental data using 1∶1 binding model with global Rmax.
Table 1.
Interaction of BL-7010 (Polymer A) with gliadin, BSA, proteolytic enzymes and vitamins.
Figure 2.
Administration of BL-7010 (Polymers A and B) at a gluten: BL-7010 ratio of 1∶3 decreased gluten-associated histological pathology in the small intestine of sensitized mice.
Small intestinal H&E stained sections from (A) non-sensitized mice (control) (B) gliadin-sensitized mice (gluten) (C) gliadin-sensitized mice, receiving Polymer A (D) gliadin-sensitized mice, receiving Polymer B. (E) non-sensitized mice (control) at different magnification (F) sensitized mice at different magnification (G) Quantification of V/C ratios from small intestinal villi. H&E sections were viewed via optical microscopy (4x and 10x magnification). Bars represent the mean +/− SEM, statistical analysis was completed via one-way ANOVA and Bonferroni's post-hoc test (*p<0.05, ****p<0.0001) (n = 12–13).
Figure 3.
Administration of BL-7010 (Polymers A and B) decreased CD3+ intraepithelial cells in villi tips of gliadin-sensitized mice.
Small intestinal CD3+ stained small intestinal tissues from (A) non-sensitized mice (control) (B) gliadin-sensitized mice (gluten) (C) gliadin-sensitized mice, receiving Polymer A (D) gliadin-sensitized mice, receiving Polymer B. (E) Quantification of CD3+ cells/100 enterocytes in villi tips. Stained sections were viewed via optical microscopy (20X magnification). Bars represent the mean +/− SEM, statistical analysis was completed via one-way ANOVA and Bonferroni's post-hoc test (***p<0.001, ****p<0.0001) (n = 11–13).
Figure 4.
Administration of BL-7010 (Polymers A and B) decreased paracellular permeability and PAT-1 mRNA levels in gliadin-sensitized mice.
(A) Gliadin-sensitized mice had higher 51Cr-EDTA flux in comparison to non-sensitized control mice, and administration of Polymers A or B decreased it to the same level as the non-sensitized control mice. (B) Gliadin-sensitized mice had higher mRNA levels of PAT-1 in comparison to non-sensitized control mice, and administration of Polymers A or B to sensitized mice decreased expression to the same level as non-sensitized control mice. Bars represent the mean +/− SEM, statistical analysis was completed via one-way ANOVA and Bonferroni's post-hoc test (**p<0.01, ***p<0.001, ****p<0.0001) (n = 12–13).