Journal
CELL TRANSPLANTATION
Volume 23, Issue 1, Pages 59-72Publisher
SAGE PUBLICATIONS INC
DOI: 10.3727/096368912X659880
Keywords
Islet; Basement membrane (BM); Vascular endothelial cells (VECs); Transplantation; Isograft; Allograft; Rejection
Funding
- National Health and Medical Research Council of Australia (NH&MRC)/JDRF Special Program Grant [418138]
- Diabetes Australia Research Trust Apex Award
- NHMRC [455395]
- Roche Organ Transplantation Research Foundation (ROTRF)/JDRF [477554991]
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The isolation of islets by collagenase digestion can cause damage and impact the efficiency of islet engraftment and function. In this study, we assessed the basement membranes (BMs) of mouse pancreatic islets as a molecular biomarker for islet integrity, damage after isolation, and islet repair in vitro as well as in the absence or presence of an immune response after transplantation. Immunofluorescence staining of BM matrix proteins and the endothelial cell marker platelet endothelial cell adhesion molecule-1 (PECAM-1) was performed on pancreatic islets in situ, isolated islets, islets cultured for 4 days, and islet grafts at 3-10 days posttransplantation. Flow cytometry was used to investigate the expression of BM matrix proteins in isolated islet 0-cells. The islet BM, consisting of collagen type IV and components of Engelbreth Holm S warm (EHS) tumor laminin 111, laminin a2, nidogen-2, and perlecan in pancreatic islets in situ, was completely lost during islet isolation. It was not reestablished during culture for 4 days. Peri- and intraislet BM restoration was identified after islet isotransplantation and coincided with the migration pattern of PECAM-1+ vascular endothelial cells (VECs). After islet allotransplantation, the restoration of VEC-derived peri-islet BMs was initiated but did not lead to the formation of the intraislet vasculature. Instead, an abnormally enlarged peri-islet vasculature developed, coinciding with islet allograft rejection. The islet BM is a sensitive biomarker of islet damage resulting from enzymatic isolation and of islet repair after transplantation. After transplantation, remodeling of both peri- and intraislet BMs restores 0-cell matrix attachment, a recognized requirement for 13-cell survival, for isografts but not for allografts. Preventing isolation-induced islet BM damage would be expected to preserve the intrinsic barrier function of islet BMs, thereby influencing both the effector mechanisms required for allograft rejection and the antirejection strategies needed for allograft survival.
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