Intestinal M cells in Peyer's patches, the specialized antigen-sampling cells of the mucosal immune system, are exploited by Salmonella and other pathogens as a route of invasion. Thus, M cells have attracted lots of attention as a major target of the mucosal immune system. Here, we report that caveolin-1 plays a crucial role in the entry of Salmonella into M cells. We established an in vitro M-like cell model in which polarized enterocyte-like Caco-2 cells created after co-culturing with the Raji B cell line that underwent a phenotypic switch to a form that morphologically and functionally resembles the specialized antigen-transporting M cells. Caveolin-1 was highly expressed in the M-like cells, while not in Caco-2 cells, and a great number of Salmonella infected caveolin-1 -expressing M-like cells. To elucidate the role of caveolin-1 in the entry of Salmonella, we downregulated caveolin-1 expression by siRNA and analyzed the level of Salmonella transcytosis across the M-like cells. Transcytosis of Salmonella was markedly reduced by downregulation of caveolin-1 in the M-like cells. These results suggest that caveolin-1 is implicated in the gateway of microbial pathogens through M cells, and, thus, provides a new target of mucosal immunity. Crown Copyright (C) 2009 Published by Elsevier Inc. All rights reserved.