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. 2014 Jul 17;63(8):2727–2743. doi: 10.2337/db13-1658

Figure 5.

Figure 5

Staining pattern of the HA-binding proteins IαI, versican, and TSG-6 in normal and diabetic islets and insulitis. A–M: Immunohistochemistry for IαI and morphometric quantification of the IαI-positive islet area. A–D: Immunohistochemistry reveals IαI in normal (A and B) and diabetic (C and D) islets. B and D: Images show higher magnification of the tissue areas within the white boxes in A and C. Blue arrowheads point to the islet border. The magenta arrowhead points to an intraislet capillary. E–H: Colabeling of IαI (red) with HA (green) indicates that IαI occurs in the HA-rich regions along the islet microvessels. Nuclei (blue) are visualized with DAPI. F–H: Images show higher magnification of the boxed area in E. I: Mean ± SD of peri- and intraislet IαI-positive areas obtained from tissues of 17 healthy and 19 T1D donors. J–L: Relative IαI-positive islet area showing mean ± SD (J) and individual values (K) of measurements obtained from tissues of 17 healthy and 19 T1D donors. L: Mean ± SD of measurements obtained from tissues of 17 healthy donors and 19 T1D donors with diabetes for ≤9 years (n = 15) and ≥28 years (n = 4). Blue bars and △, normal tissues; red bars and ○, diabetic tissues. *P < 0.0001 vs. normal tissues. M: Image shows higher magnification of the tissue area within the yellow box in C. Immunohistochemistry reveals IαI accumulation in insulitis. Scale bars: 50 μm (A, C, and E) and 10 μm (B, D, F–H, and M). N–S: Immunohistochemistry for versican and morphometric quantification of the intraislet versican-positive area. N: Immunohistochemistry shows that versican is sparse in normal islets. O: Higher magnification of the boxed area in N. P: Immunohistochemistry shows that versican accumulates in diabetic islets and locates adjacent to the islet microvessels (white box) and in insulitis area (yellow box). Q and S: Higher magnification of tissue areas within the white and yellow boxes, respectively, in P. Blue arrowheads point to the islet border. Magenta and green arrowheads point to intraislet capillaries. Images were prepared from nPOD cases 6153 (healthy) and 6052 (T1D). Scale bar: 50 μm (N and P) and 10 μm (O, Q, and S). R: Morphometric quantification of the relative versican-positive islet area. Blue bar, normal tissues; red bar, diabetic tissues. Data are mean ± SD of measurements obtained from tissues of 17 healthy and 19 T1D donors. **P = 0.00154 vs. normal tissues. T–Y: Immunohistochemistry for TSG-6 and morphometric quantification of the intraislet TSG-6–positive area. T: Immunohistochemistry shows that TSG-6 occurs within the pancreatic endocrine cells in normal tissues. U: Higher magnification of the boxed area in T. V: Immunohistochemistry shows that TSG-6 is present in diabetic islets (white box) and in a subset of inflammatory cells in insulitis (yellow box). W and Y: Higher magnification of tissue areas within the white and yellow boxes, respectively, in V. Blue arrowheads point to the islet border. Magenta and green arrowheads point to intraislet capillaries. Images were prepared from nPOD cases 6153 (healthy) and 6052 (T1D). Scale bar: 50 μm (T and V) and 10 μm (U, W, and Y). X: Morphometric quantification of the relative TSG-6–positive islet area. Blue bar, normal tissues; red bar, diabetic tissues. Data are mean ± SD of measurements obtained from tissues of 17 healthy and 19 T1D donors. There was a trend toward reduced TSG-6 staining area (P = 0.0554).