Our work aimed to describe intestinal HMGB1 release after infection of gnotobiotic piglets with Gram-negative enteric pathogen Salmonella Typhimurium, signaling via TLR2, TLR4, TLR9, and RAGE, and the possible influence of previous colonization of the piglets with commensal Gram-positive lactobacilli Lactobacillus amylovorus and Lactobacillus mucosae or probiotic Gram-negative E. coli Nissle 1917. Here, TLR2 is linked to infection.