Remission of Type 1 Diabetes after Anti-CD3 Antibody Treatment and Transplantation of Embryonic Pancreatic Precursors
Endocrinology Begum et al. 150: 4512 Supplemental Data
Supplemental Data
Files in this Data Supplement:
- Supplemental Figure 1 - Ubiquitous nuclear pancreatic expression of H2B-EGFP in adult NOR-CAG::H2B-EGFP mice. Frozen sections (10 μm) of adult NOR-CAG::H2B-EGFP were triple stained with anti-GFP ( green), anti-insulin ( red) and DAPI ( blue) to confirm the nuclear GFP expression in both exocrine and endocrine tissues. DAPI and anti-GFP staining show nuclear GFP expression is in both exocrine and endocrine tissues of the pancreas. Anti-insulin antibody staining shows cytoplasmic insulin expression in the islet. Merged image of anti-insulin and anti-GFP antibody are shown on the right. Bar = 100μm.
- Supplemental Figure 2 - Mice with a low number of anlagen transplanted (2-3) (A) or initial high blood glucose (>500 mg/dl) (B) fail to maintain normal blood glucose following combination treatment with F(ab')2 anti-CD3 antibody, pancreatic anlagen, and an insulin pellet. X-axis represents days post-anlagen transplantation.
- Supplemental Figure 3 - Analysis of donor tissue in the kidney graft site. (A) Representative frozen sections of the graft site 20 days after transplantation showing donor-derived, GFP-positive cells and abundant scattered anti-insulin or anti-glucagon-positive cells. Anti-PDX1-positive cells line duct-like structures and are not insulin positive. (B) Representative frozen sections of the graft site 45 days after transplantation showing only a few scattered anti-insulin or anti-glucagon-positive cells.
- Supplemental Figure 4 - Presence of T-cells indicates some degree of periinsulitis in treated mice. (A) Immunohistochemical analysis with anti-CD3 antibody carried out on frozen sections of pancreas from diabetic NOD mice that received the combination treatment and were unilaterally nephrectomized at 45 days shows the presence of T-cells surrounding some but not all of the graft-derived, GFP-positive areas. (B) Similarly, endogenous islets in diabetic treated mice show a range in the number of surrounding T-cells. Bar = 100μm.