We recently discovered a structurally novel class of endogenous lipids, branched palmitic acid esters of hydroxy stearic acids (PAHSAs), with beneficial metabolic and anti-inflammatory effects. We tested whether PAHSAs protect against colitis, which is a chronic inflammatory disease driven predominantly by defects in the innate mucosal barrier and adaptive immune system. There is an unmet clinical need for safe and well tolerated oral therapeutics with direct anti-inflammatory effects. Wild-type mice were pretreated orally with vehicle or 5-PAHSA (10 mg/kg) and 9-PAHSA (5 mg/kg) once daily for 3 days, followed by 10 days of either 0% or 2% dextran sulfate sodium water with continued vehicle or PAHSA treatment. The colon was collected for histopathology, gene expression, and flow cytometry. Intestinal crypt fractions were prepared for ex vivo bactericidal assays. Bone marrow-derived dendritic cells pretreated with vehicle or PAHSA and splenic CD4 ؉ T cells from syngeneic mice were co-cultured to assess antigen presentation and T cell activation in response to LPS. PAHSA treatment prevented weight loss, improved colitis scores (stool consistency, hematochezia, and mouse appearance), and augmented intestinal crypt Paneth cell bactericidal potency via a mechanism that may involve GPR120. In vitro, PAHSAs attenuated dendritic cell activation and subsequent T cell proliferation and Th1 polarization. The anti-inflammatory effects of PAHSAs in vivo resulted in reduced colonic T cell activation and pro-inflammatory cytokine and chemokine expression. These anti-inflammatory effects appear to be partially GPR120-dependent. We conclude that PAHSA treatment regulates innate and adaptive immune responses to prevent mucosal damage and protect against colitis. Thus, PAHSAs may be a novel treatment for colitis and related inflammation-driven diseases.Lipids have been a rich source of chemical matter for the development of new medicines (1). Endogenous small-molecule metabolites are another class of natural products made within the human body that have important biological activities (2). For example, the discovery and synthesis of prostaglandins revealed their diverse biological roles as autocrine and paracrine mediators in many tissues. This led to their development as therapeutic agents for a variety of indications ranging from vasodilatation to glaucoma treatment (3-6). More recently, compounds that target the sphingosine 1 receptors are emerging as novel therapeutics for autoimmune diseases (7). Thus, the discovery and characterization of bioactive lipids is of fundamental interest with broad impact. We recently discovered a structurally novel class of lipids, branched fatty acid esters of hydroxy fatty acids (FAHFAs), 6 with anti-diabetic and anti-inflammatory properties (8). These lipids are products of endogenous synthesis in mammalian tissues and are also present in food. There are more than 16 FAHFA families, and each family is distinguished by having a different fatty acid and hydroxy fatty acid composition. Furthermor...