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Infection and Immunity, March 2006, p. 1555-1564, Vol. 74, No. 3
0019-9567/06/$08.00+0     doi:10.1128/IAI.74.3.1555-1564.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

The Outer Core Lipopolysaccharide of Salmonella enterica Serovar Typhi Is Required for Bacterial Entry into Epithelial Cells

Anilei Hoare,1 Mauricio Bittner,1 Javier Carter,1 Sergio Alvarez,1 Mercedes Zaldívar,1 Denisse Bravo,1 Miguel A. Valvano,2 and Inés Contreras1*

Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias Químicas y Farmacéuticas, Universidad de Chile, P.O. Box 174, Correo 22, Santiago, Chile,1 Department of Microbiology and Immunology, Siebens-Drake Research Institute, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, Ontario N6A 5C1, Canada2

Received 29 July 2005/ Returned for modification 26 October 2005/ Accepted 22 December 2005

Salmonella enterica serovar Typhi causes typhoid fever in humans. Central to the pathogenicity of serovar Typhi is its capacity to invade intestinal epithelial cells. The role of lipopolysaccharide (LPS) in the invasion process of serovar Typhi is unclear. In this work, we constructed a series of mutants with defined deletions in genes for the synthesis and polymerization of the O antigen (wbaP, wzy, and wzz) and the assembly of the outer core (waaK, waaJ, waaI, waaB, and waaG). The abilities of each mutant to associate with and enter HEp-2 cells and the importance of the O antigen in serum resistance of serovar Typhi were investigated. We demonstrate here that the presence and proper chain length distribution of the O-antigen polysaccharide are essential for serum resistance but not for invasion of epithelial cells. In contrast, the outer core oligosaccharide structure is required for serovar Typhi internalization in HEp-2 cells. We also show that the outer core terminal glucose residue (Glc II) is necessary for efficient entry of serovar Typhi into epithelial cells. The Glc I residue, when it becomes terminal due to a polar insertion in the waaB gene affecting the assembly of the remaining outer core residues, can partially substitute for Glc II to mediate bacterial entry into epithelial cells. Therefore, we conclude that a terminal glucose in the LPS core is a critical residue for bacterial recognition and internalization by epithelial cells.

* Corresponding author. Mailing address: Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias Químicas y Farmacéuticas, Universidad de Chile, P.O. Box 174, Correo 22, Santiago, Chile. Phone: (56-2) 978 1658. Fax: (56-2) 222 7900. E-mail: icontrer{at}uchile.cl.

Editor: V. J. DiRita

Infection and Immunity, March 2006, p. 1555-1564, Vol. 74, No. 3
0019-9567/06/$08.00+0     doi:10.1128/IAI.74.3.1555-1564.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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