Household endotoxin levels have been variably associated with risk for asthma and atopy.We studied participants from the 2005-2006 National Health and Nutrition Examination Survey (NHANES, n=6963), a large cohort representative of the US population (age 1-84 years). We built logistic regression models to test for associations between house dust endotoxin and sensitization to specific foods (milk, egg, and peanut). To experimentally explore the detected epidemiologic associations, peripheral blood mononuclear cells (PBMCs) were collected from 21 children (age 1-19 years) mono-food allergic (i.e. sensitized and clinically reactive) to milk, egg, or peanut and non-allergic controls for stimulation with endotoxin and secreted cytokine measurement. For each food allergy, linear mixed effects models were built to test the association between endotoxin stimulation and cytokine level.Among NHANES subjects, the geometric mean household endotoxin level was 15.5 EU/mg (GSE 0.5). Prevalence of food allergen sensitization (sIgE≥0.35 kU /L) varied by food: milk 5.7%, egg 4.0%, and peanut 7.9%. In models adjusted for potential confounders (age, race, country of birth, total people per household, US region, and history of wheezing in the past year), household endotoxin level was associated with sensitization to milk (OR 1.7, 95%CI 1.2-2.1) and egg (OR 1.4, 95%CI 1.01-1.9), but not peanut (OR 0.98, 95%CI 0.8-1.2). Interferon-γ levels of endotoxin-stimulated PBMCs from children allergic to milk or egg, but not peanut, were significantly lower compared to controls in linear mixed effects models adjusted for repeated measures, experimental variables, age, and inter-individual variability (P-values 0.007, 0.018, and 0.058, respectively,).Higher household endotoxin is associated with increased odds of milk and egg sensitization. Altered cytokine responsiveness to endotoxin is also observed in PBMCs from individuals with milk and egg allergy.