Acetylated-lysine antibody was purchased from Cell Signaling Technology

well as therapeutic PG-490 strategies for restoring normal autophagy in cystic fibrosis patients will be an 7621916 essential step towards improved treatment of P. aeruginosa infection. Excitingly, recent work has identified a peptide fragment from beclin-1 which strongly and specifically induces autophagy, and was able to decrease mortality associated with viral pathogens in vivo. Such targeted therapeutic strategies for the modulation of autophagy have considerable clinical potential for the effective control and irradiation of P. aeruginosa lung infections. Intracellular pathogens have developed a wide range of mechanisms for subverting or usurping the host autophagy pathway to promote their pathogenicity and survival. Given the novel observations that autophagy contributes to the clearance of P. aeruginosa in vivo, it raises the possibility that the bacteria have evolved similar mechanisms to avoid killing through the autophagy pathway. P. aeruginosa bacteria employs various mechanisms which skew the host immune response towards a Th2 phenotype. It has long been thought that a Th2 like response is favorable in the clearance of extracellular infections, while a Th1 response favors killing of intracellular infections. However, in the 2173565 case of P. aeruginosa infections a Th2-like response is in fact correlated with poorer lung function and clinical outcomes compared to the minority of patients which mount a Th1-like response, underscoring the importance of the intracellular phase of P. aeruginosa infection. Interestingly, Th2 cytokines have been demonstrated to suppress autophagy, suggesting that the Th2 responses driven by P. aeruginosa could promote the bacteria’s intracellular survival by preventing activation of the autophagy pathway. In addition, P. aeruginosa secretes exotoxin A which shuts down host protein synthesis through inhibiting eukaryotic elongation factor 2A which greatly enhances the virulence of the bacteria. However disruption of the eEF2A signaling pathways has also been shown to impair activation of autophagy. Thus, P. aeruginosa may employ multiple mechanisms for disrupting the autophagy pathway, contributing to enhanced bacterial survival and pathogenicity. Given the increasing prevalence of multi-antibiotic resistant strains of P. aeruginosa bacteria, novel therapeutic approaches for the treatment of chronic infections in individuals with cystic fibrosis will be essential to ensure the continued health of these patients in the coming years. Here we demonstrate that P. aeruginosa induces autophagy in mast cells, and that the pathway contributes to the killing of internalized bacteria in vitro. We further demonstrate that therapeutic intervention aimed at inducing autophagy with rapamycin correlates with decreased bacterial loads following P. aeruginosa lung infection in vivo, in spite of decreased inflammatory responses and neutrophil infiltration caused by rapamycin treatment. This work provides the first evidence that the pharmacological manipulation of the autophagy pathway has considerable therapeutic potential in the treatment of P. aeruginosa infections in vivo. The thick coat of carbohydrates in the glycocalyx that emerges from apical membranes of epithelial cells is critical to maintaining barrier function on mucosal surfaces. This glycocalyx is important in preventing access of microbes to plasma membranes, but also significantly restricts drug and vaccine targeting of epithelial cells. In the eye, the bioavailability