Methods to Study TLRs in Transplantation.

Toll-like receptors (TLRs) are key regulators of immune responses, including alloimmune responses. In this chapter, we present protocols to study whether and/or how TLRs can contribute to solid-organ transplant rejection. We describe methods to reduce heterogeneity in microbiome variations between animals before beginning experiments to limit confounding factors, protocols using TLR agonists to prevent anti-CD154/donor splenocyte transfer-mediated tolerance, and recipes to heat-kill microbes or use hosts genetically deficient in TLR-dependent pathways to distinguish between TLR-dependent and live bacteria-dependent effects.