ABSTRACT
We provide the first comprehensive analysis of the extracellular matrix (ECM) composition of peri-islet capsules, composed of the peri-islet basement membrane (BM) and subjacent interstitial matrix (IM), in development of type 1 diabetes in NOD mice and in human type 1 diabetes. Our data demonstrate global loss of peri-islet BM and IM components only at sites of leukocyte infiltration into the islet. Stereological analyses reveal a correlation between incidence of insulitis and the number of islets showing loss of peri-islet BM versus islets with intact BMs, suggesting that leukocyte penetration of the peri-islet BM is a critical step. Protease- and protease inhibitor-specific microarray analyses (CLIP-CHIP) of laser-dissected leukocyte infiltrated and noninfiltrated pancreatic islets and confirmatory quantitative real time PCR and protein analyses identified cathepsin S, W, and C activity at sites of leukocyte penetration of the peri-islet BM in association with a macrophage subpopulation in NOD mice and human type 1 diabetic samples and, hence, potentially a novel therapeutic target specifically acting at the islet penetration stage. Interestingly, the peri-islet BM and underlying IM are reconstituted once inflammation subsides, indicating that the peri-islet BM-producing cells are not lost due to the inflammation, which has important ramifications to islet transplantation studies.
Subject(s)
Basement Membrane/immunology , Chemotaxis, Leukocyte/immunology , Diabetes Mellitus, Type 1/immunology , Extracellular Matrix/immunology , Islets of Langerhans/immunology , Animals , Cathepsin C/analysis , Cathepsin W/analysis , Cathepsins/analysis , Humans , Inflammation/metabolism , Islets of Langerhans/pathology , Macrophages/immunology , Macrophages/pathology , Mice , Mice, Inbred NOD , Protein Array Analysis , Proteinase Inhibitory Proteins, Secretory/analysisABSTRACT
Cathepsin W is exclusively expressed in immune cells, and a novel isoform was identified previously. To characterize the expression pattern of the wildtype and isoform Ins10, specific polymerase chain reaction assays were generated and used to study respective transcript levels in peripheral blood cells and gastric biopsies in healthy subjects. The wildtype-encoding transcript levels were 3- and 9-fold higher in mucosal samples and peripheral immune cells, respectively (p<0.05). The predominant expression of wildtype form by infiltrating immune cells was confirmed in 116 patients with gastroesophageal reflux disease and 27 reflux-negative individuals demonstrating that cathepsin W expression is not altered in this disease.
