The islet mass is already marginal shortly after transplantation and thus susceptible to become insufficient when subsequently exposed to negative local influences. Recent estimates indicate that less than 30% of islets stably engraft, a result

that explains the requirement for infusing large numbers of islets and for repeat islet infusions to maintain insulin-free euglycemia 2. Mechanisms underlying early islet loss following transplantation remain poorly defined but apoptotic cell islet cell death associated with peri- and intra-islet graft inflammation have been described previously 3, 4. TLR are a family of pattern recognition receptors that bind to PAMP or to endogenous ligands released Y 27632 by damaged cells (damage-associated molecular patterns, DAMP). Among the latter group are HSPs, high-mobility group box protein 1 (HMGB1), heparan sulfate, hyaluronan fragments, and fibronectin 5. Regardless MI-503 mouse of the source of the

specific ligand, TLR-transmitted signals activate innate immunity by inducing chemokine and cytokine release and through upregulating costimulatory molecule expression, among a multitude of other effects 6. Recent studies revealed the importance of islet-expressed TLR, particularly TLR2 and TLR4, participating in the pathogenesis of autoimmune diabetes and allogeneic islet transplant rejection 7–9. Whether TLR transmitted signals in the islets impact early islet engraftment has not been studied. Our group, among others, showed that following physical manipulation, prolonged cell culture, ischemia/reperfusion injury, or virus-mediated

gene transduction, islets can produce cytokines and chemokines in patterns reminiscent Baricitinib of those induced by TLR stimulation 10–15. Upon transplantation, such manipulations amplify peri-islet inflammation and result in impaired islet graft function, further supporting the concept that early islet injury is in part mediated through TLR signals. To define the mechanisms of early graft dysfunction, we studied the impact of TLR stimulation on graft survival following transplantation. Our data provide the first direct evidence that islet-expressed TLR2 and TLR4 are relevant mediators of the post-transplant inflammation associated with early graft dysfunction. These effects require recipient T cells, occur in the absence of islet DC, and are fully reproduced by stimulation with HMGB1, an endogenous TLR2/4 ligand that is released by pancreatic tissue after sterile injury. In addition to providing insight into mechanisms underlying early graft loss, our findings indicate that TLR2 and TLR4 are potential targets for novel therapies aimed at preserving islet mass. Using RT-PCR, we found that RNA from a pancreatic β cell line and from purified C57BL/6 islets expressed message for TLR2 and TLR4 (Fig. 1A).