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Dendritic cells (DCs) are key mediators of the innate response to transplantation. Yet, the substances that activate these cells during acute allograft rejection remain elusive. Previous work has suggested that heat shock protein (HSP)-70 is associated with acute allograft rejection. Hence, the goal of this study was to determine whether HSP-70 activates DCs and plays a critical role in acute allograft rejection in an experimental model that is dependent on innate MyD88 signaling. Our in vitro data indicate that HSP-70 does not activate DCs. In vivo transplant studies demonstrate that HSP-70 levels are not increased during acute allograft rejection and that an absence of the inducible form of HSP-70 neither delays acute allograft rejection, impairs DCs maturation, nor alters Th1 immune responses during acute allograft rejection. In conclusion, our results indicate that HSP-70 in our experimental models does not play an essential role in acute allograft rejection.