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© The Rockefeller University Press, 0021-9525/1999//689 $5.00
The Journal of Cell Biology, Volume 145, Number 4, , 1999 689-698

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Trafficking of Shigella Lipopolysaccharide in Polarized Intestinal Epithelial Cells



Wandy L. Beatty*, Stéphane Méresse{ddagger}, Pierre Gounon§, Jean Davoust{ddagger}, Joëlle Mounier*, Philippe J. Sansonetti*, and Jean-Pierre Gorvel{ddagger}

* Unité de Pathogénie Microbienne Moléculaire, U389, Institut National de la Santé et de la Recherche Médicale, Institut Pasteur, 75724 Paris Cédex 15, France; {ddagger} Centre d'Immunologie-Marseille-Luminy, Parc Scientifique et Technologique de Luminy, Case 906, 13288 Marseille Cédex 9, France; and § Station Centrale de Microscopie Electronique, Institut Pasteur, 75724 Paris Cédex 15, France

Bacterial lipopolysaccharide (LPS) at the apical surface of polarized intestinal epithelial cells was previously shown to be transported from the apical to the basolateral pole of the epithelium (Beatty, W.L., and P.J. Sansonetti. 1997. Infect. Immun. 65:4395–4404). The present study was designed to elucidate the transcytotic pathway of LPS and to characterize the endocytic compartments involved in this process. Confocal and electron microscopic analyses revealed that LPS internalized at the apical surface became rapidly distributed within endosomal compartments accessible to basolaterally internalized transferrin. This compartment largely excluded fluid-phase markers added at either pole. Access to the basolateral side of the epithelium subsequent to trafficking to basolateral endosomes occurred via exocytosis into the paracellular space beneath the intercellular tight junctions. LPS appeared to exploit other endocytic routes with much of the internalized LPS recycled to the original apical membrane. In addition, analysis of LPS in association with markers of the endocytic network revealed that some LPS was sent to late endosomal and lysosomal compartments.

Key Words: Shigella • lipopolysaccharide • transcytosis • trafficking • epithelial

Abbreviations used in this paper: CI-MPR, cation-independent mannose 6-phosphate receptor; FCS, flow cytometry standard; LAL, limulus amebocyte lysate assay; Lamp, lysosomal-associated membrane protein; LPS, lipopolysaccharide; MIIC, MHC class II.


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