The ability of natural tolerance to be applied to allogeneic tissue: determinants and limits.

Background: Transplant rejection has been considered to occur primarily because donor antigens are not present during the development of the recipient's immune system to induce tolerance. Thus, transplantation prior to recipient immune system development (pre-immunocompetence transplants) should induce natural tolerance to the donor. Surprisingly, tolerance was often not the outcome in such 'natural tolerance models'.

Non-myeloablative mixed chimerism approaches and tolerance, a split decision.

Stable mixed chimerism has been considered the most robust tolerance strategy. However, rejection of solid donor tissues by chimeras has been observed, a state termed split tolerance. Since new non-myeloablative mixed chimerism approaches are being actively pursued, we sought to determine whether they lead to full tolerance or split tolerance and to define the mechanisms involved.

Effect of prolonged in vitro exposure to high glucose on neonatal porcine pancreatic islets.

Prolonged exposure to high glucose can influence the function, growth, and survival of pancreatic beta-cells. In this study, we examine the effects of prolonged in vitro exposure to high glucose on neonatal porcine beta-cells, a potentially useful source of insulin-producing cells for clinical islet transplantation. Neonatal porcine islets were prepared by culturing collagenase-digested pancreases for 1 week in 5.

Immune mechanisms associated with the rejection of encapsulated neonatal porcine islet xenografts.

Background: The immune mechanisms associated with the rejection of microencapsulated neonatal porcine islets (NPI) are not clearly understood. Therefore, in this study we characterized the immune cells and molecules that are involved in this process by examining the microencapsulated NPI xenografts at various time points post-transplantation in B6 mice.

Methods: Microencapsulated NPI were transplanted into streptozotocin-induced diabetic immune-competent B6 and immune-deficient B6 rag-/- mice and blood glucose levels were monitored twice a week.

The impact of the mTOR inhibitor sirolimus on the proliferation and function of pancreatic islets and ductal cells.

Aims/hypothesis: The Edmonton Protocol for islet transplantation has provided hope for type 1 diabetic patients. However, this protocol requires lifelong immunosuppression, specifically sirolimus, a cellular antiproliferate. The effect of sirolimus on human pancreatic ductal cells (HDCs) is not known.

Interventional strategies to prevent beta-cell apoptosis in islet transplantation.

A substantial proportion of the transplanted islet mass fails to engraft due to death by apoptosis, and a number of strategies have been explored to inhibit beta-cell loss. Inhibition of extrinsic signals of apoptosis (i.e.

Ectopic expression of neurogenin 3 in neonatal pig pancreatic precursor cells induces (trans)differentiation to functional alpha cells.

Aims/hypothesis: Neurogenin 3 (NEUROG3), a basic helix-loop-helix transcription factor that is needed for endocrine cell development in the embryonic pancreas, has been shown to induce transdifferentiation of duct cells from adult pancreas towards a neuro-endocrine phenotype. Our study explored the endocrine transdifferentiation potential of NEUROG3 in neonatal pancreatic precursor cells.

Materials And Methods: A replication-deficient adenovirus expressing Neurog3 and green fluorescent protein (GFP) (Ad-NEUROG3) was used to infect neonatal pig pancreatic cell preparations enriched for endocrine islet and cytokeratin-positive precursor cells.

Ameliorating injury during preservation and isolation of human islets using the two-layer method with perfluorocarbon and UW solution.

This study assessed the effects of a two-layer method (TLM), using perfluorocarbon and UW solution, on the quality of human pancreata following storage and islet yield/function after isolation. In part A, TLM was applied immediately after procurement and the energetic profile was compared to a group treated with UW solution only (control) throughout 24-h storage. In part B, cadaveric human pancreata were procured and subjected to a TLM after cold storage in UW solution (TLM group) or UW solution (control group).

Time, space and contextual models of the immunity tolerance decision: bridging the geographical divide of Zinkernagel and Hengartner's 'Credo 2004'.

In Credo 2004, Zinkernagel and Hengartner (Z&H) have continued their challenge to the immunological community to reconsider assumptions regarding the most fundamental aspects of adaptive immunity. They have appropriately championed the role of persistent, widely distributed antigen in tolerance induction, parameters that do not figure prominently in most other models. The global theory of immunity they have developed is predominantly based on observations from studies with viruses and tumours.

Control of oxidative stress in small bowel: relevance to organ preservation.

Background: Oxidative stress during cold small bowel (SB) storage has not been investigated because oxygen is depleted rapidly after procurement. We hypothesized that oxidative catabolism facilitated by a proven amino acid-based (AA) storage solution promotes oxidative stress; furthermore, there is an important role for antioxidant supplementation during cold storage.

Methods: SB from Sprague-Dawley rats (n = 6 in each group) were procured according to standardized procedures involving vascular flush with modified University of Wisconsin solution and luminal treatment with an AA-based solution proven previously to aid preservation.

Expansion of mesenchymal stem cells from human pancreatic ductal epithelium.

Fibroblast-like cells emerging from cultured human pancreatic endocrine and exocrine tissue have been reported. Although a thorough phenotypic characterization of these cells has not yet been carried out, these cells have been hypothesized to be contaminating fibroblasts, mesenchyme and/or possibly beta-cell progenitors. In this study, we expanded fibroblast-like cells from adult human exocrine pancreas following islet isolation and characterized these cells as mesenchymal stem cells (MSCs) based on their cell surface antigen expression and ability to differentiate into mesoderm.

Nutrient-related issues affecting successful experimental orthotopic small bowel transplantation.

Background: This study tested the effectiveness of a nutrient-rich preservation solution in a small animal model of orthotopic whole small bowel transplantation.

Methods: Lewis rats received syngeneic total orthotopic small bowel graft after cold storage for 6 h. Donor small bowel was flushed vascularly with University of Wisconsin (UW) solution and flushed luminally with UW solution or an amino acid-rich (AA) solution as follows: Group 1, no luminal flush; Group 2, UW solution; Group 3, AA solution.

Development of an ectopic site for islet transplantation, using biodegradable scaffolds.

Clinical islet transplantation in liver has achieved normoglycemia. However, this site may not be ideal for islet survival. To create a more optimal site for islet transplantation, we have developed a construct with biodegradable scaffolds.

Prolonged survival of microencapsulated neonatal porcine islets in mice treated with a combination of anti-CD154 and anti-LFA-1 monoclonal antibodies.

Background: The aim of this study was to determine whether short-term administration of a combination of anti-CD154 and anti-LFA-1 monoclonal antibodies can prolong the survival of microencapsulated neonatal porcine islets (NPI) in immunocompetent mice.

Methods: Microencapsulated NPI were transplanted into the peritoneal cavity of streptozotocin-induced diabetic B6 mice that received a short-term treatment of a combination of anti-CD154 and anti-LFA-1 monoclonal antibodies. Blood glucose levels of each recipient were measured for more than 100 days posttransplantation or until graft rejection.

Quantitative assessment of collagenase blends for human islet isolation.

Background: The variability in collagenase blends has been speculated as the single most important determinant of the success or failure in isolated islet yields in clinical islet transplantation. Examination of the formulation and potency of the widely used Liberase HI enzyme blend will uncover possible sources of imprecision.

Methods: High performance liquid chromatography (HPLC) and kinetic measurements of collagenase and protease activity were used to assess potency.

Robust tolerance to fully allogeneic islet transplants achieved by chimerism with minimal conditioning.

Background: Whether mixed chimeras induced by nonmyeloablative conditioning are tolerant to challenge with donor allogeneic islet grafts is unknown. Here we investigate whether our nonmyeloablative, costimulation blockade-free and sirolimus (SRL)-based protocol could facilitate mixed chimerism via bone marrow transplantation (BMT) and induce islet allograft tolerance.

Methods: After low dose (1-3 Gy) total body irradiation (TBI, day -1), with or without prior lymphocyte depletion, C57BL/6 mice were transfused with 40 x 10(6) BALB/c bone marrow cells (day 0) and received SRL (3 mg/kg/day) for 4 weeks.

Sildenafil improves alveolar growth and pulmonary hypertension in hyperoxia-induced lung injury.

Rationale: Bronchopulmonary dysplasia (BPD), the chronic lung disease of preterm infants, and pulmonary emphysema, both significant global health problems, are characterized by an arrest in alveolar growth/loss of alveoli structures. Mechanisms that inhibit distal lung growth are poorly understood, but recent studies suggest that impaired vascular endothelial growth factor signaling and reduced nitric oxide (NO) production decreases alveolar and vessel growth in the developing lung, features observed in experimental oxygen-induced BPD. NO exerts its biological activity by stimulating guanosine 3',5'-cyclic monophosphate (cGMP) production.

The standardization of pancreatic donors for islet isolation.

Introduction: Islet transplantation has proven to be a successful treatment for insulin-dependent diabetes mellitus (IDDM). The aim of this study was to establish an algorithm by which the combination of the donor quality and pancreas quality was given a numerical score from 0 to 100 for use in determining the quality of a pancreas for islet isolation.

Methods: In this study we retrospectively analyzed 326 pancreata and the outcomes of their respective isolations.

The influence of short-term fasting on the quality of small bowel graft preservation.

Introduction: Donor nutritional status may be a determinant of small bowel (SB) quality following storage. In this study, we investigated the effect of donor nutritional status and a proven nutrient-rich preservation solution on graft quality following cold storage.

Methods: Rats were fasted (12-14 h) or non-fasted.

Indefinite survival of neonatal porcine islet xenografts by simultaneous targeting of LFA-1 and CD154 or CD45RB.

A variety of transient therapies directed against molecules involved in T-cell activation and function result in long-term islet allograft survival. However, there are relatively few examples of durable islet xenograft survival using similar short-term approaches, especially regarding highly phylogenetically disparate xenograft donors. Previous studies demonstrate that combined anti-lymphocyte function-associated antigen-1 (LFA-1) plus anti-CD154 therapy results in a robust form of islet allograft tolerance not observed with either individual monotherapy.

Neonatal porcine Sertoli cells inhibit human natural antibody-mediated lysis.

Sertoli cells protect cotransplanted cells from allogeneic and xenogeneic rejection. Additionally, neonatal porcine Sertoli cells (NPSCs) survive long-term as xenografts in nonimmunosuppressed rodents. This has led to the hypothesis that NPSCs could be used to prevent cellular rejection in clinical transplantation, thereby eliminating the need for chronic immunosuppression.

Islet graft assessment in the Edmonton Protocol: implications for predicting long-term clinical outcome.

The success of the Edmonton Protocol for islet transplantation has provided new hope in the treatment of type 1 diabetes. This study reports on the assessment of 83 human islet grafts transplanted using the Edmonton Protocol since 1999. Cellular composition, as assessed by immunohistochemistry, showed a lower islet purity (approximately 40%) than has been reported in previous studies using dithizone staining to quantitate islet equivalents.

Improved survival of microencapsulated islets during in vitro culture and enhanced metabolic function following transplantation.

Aims/hypothesis: The aim of this study was to determine whether a simple alginate capsule can prolong islet survival and function during long-term tissue culture. We also wanted to observe the ability of these encapsulated islets to restore glucose responsiveness to diabetic recipients, along with the quantity of islets required to do so.

Methods: We compared the recovery and metabolic function of encapsulated canine islets with that of non-encapsulated canine islets following 1, 2 or 3 weeks of tissue culture.

Reassessment of the vascularization of renal subcapsular islet grafts.

Objectives: The purpose of this study was to evaluate the functional significance of the renal capsular vessels in renal subcapsular islet grafts.

Methods: Syngeneic islets were transplanted under the left renal capsule of diabetic Wistar-Furth rats. At 4 weeks post-transplantation, the left renal artery and vein were ligated (group A); in group B, the left renal capsule was dissected to remove capsular vessels; or both were performed in group C.

Mechanisms and models of peripheral CD4 T cell self-tolerance.

Each response made by our immune system is either to promote or to prevent immune reactivity. That the immune system can successfully achieve both goals under specific biological conditions depends on central and peripheral tolerance mechanisms. Over the years, various experimental systems have been generated to study peripheral CD4 T cell tolerance.