Microbial strategies to target, cross or disrupt epithelia

Sandra Sousa1,2, Marc Lecuit1,3 and Pascale Cossart1

Epithelia are highly organized structures adapted to protect the
underlying tissues from external aggressions, including
microbial infections. Consequently, pathogens have evolved
various strategies to target directly or indirectly intercellular
junctions and/or components that maintain the structure of
epithelia. Interestingly, some extracellular pathogens secrete
enzymes that modify the extracellular part of junction
components. Others produce toxins that are endocytosed and
act from the inside of the cell to disrupt epithelial junctions.
Other pathogens may directly inject into cells factors that are
targeted to and destabilize the junctions, or that interact with
signaling cascades that affect junction stability. Finally invasive
bacteria or viruses may, by entering into cells, destabilize the
junctions by targeting junction components directly or by
inducing a series of events that lead to chemokine secretion,
polymorphonuclear recruitment and inflammation.
Addresses
1
Unite des Interactions Bacteries-Cellules Institut Pasteur, INSERM
U604, INRA USC2020, 28 Rue du Dr Roux, 75724 Paris Cedex 15,
France
2
Institute for Molecular and Cell Biology, Molecular Microbiology,
Rua do Campo Alegre, 823, 4150-180 Porto, Portugal
3
Service des Maladies Infectieuses et Tropicales, Hopital NeckerEnfants malades, 149 rue de Se`vres, 75015 Paris, France
Corresponding author: Cossart, P (pcossart@pasteur.fr)

Current Opinion in Cell Biology 2005, 17:489498

This review comes from a themed issue on
Cell-to-cell contact and extracellular matrix
Edited by Inke S Nathke and W James Nelson
Available online 15th August 2005
0955-0674/$ see front matter
# 2005 Elsevier Ltd. All rights reserved.
DOI 10.1016/j.ceb.2005.08.013

Introduction
A monolayered epithelium is a highly organized structure
made of polarized epithelial cells. These cells have an
apical and a basolateral pole, each characterized by a
distinct composition of proteins and lipids, defining different membrane domains with specific functions in
barrier formation and maintenance. In the intestinal
epithelium, the apical surface is made of microvilli that
constitute the exchange surface where nutrients and
fluids are absorbed and where immune system effectors
are secreted. The basolateral pole sits on a basement
membrane in contact with connective tissue, vessels and
cellular effectors of the immune system, and is responsible for the establishment of cellcell and cellmatrix
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contacts critical for maintaining epithelial polarization

and impermeability [1]. Three specialized structures,
tight junctions (TJs), adherens junctions (AJs) and desmosomes, attach epithelial cells to each other. Focal
contacts connect the basolateral surface of epithelial cells
to the extracellular matrix (Figure 1a).
TJs are the most apical structures of cellcell junctions
and define the boundary between the apical and basolateral domains. They are composed of transmembrane
proteins (such as occludins, claudins and junctional adhesion molecules or JAMs) and intracellular proteins (such
as ZO-1, -2, -3, MAGUK and PAR family proteins). TJs
act as bolts preventing paracellular diffusion and seal the
intercellular space between adjacent cells, thus creating a
barrier against free diffusion of fluids, electrolytes and
macromolecules as well as of luminal microorganisms and
their secreted products. TJs are crucial for the development and maintenance of epithelial polarity as they also
prevent diffusion of plasma membrane components from
the apical to the basolateral domains and vice versa [2].
AJs, which are required for the integrity of the TJs, are
subjacent to the TJs and are made mostly of E-cadherin, a
transmembrane protein involved in homophilic interactions. Through its cytoplasmic tail, E-cadherin binds
intracellular proteins called catenins, which interact with
the actin cytoskeleton and recruit several other proteins
involved in the development and regulation of intercellular junctions. E-cadherin interaction with the actin
cytoskeleton via catenins is necessary for full E-cadherin
adhesive activity [3,4].
Desmosomes are intercellular adhesive structures present
in epithelial cells and abundant in the skin and the
gastrointestinal mucosa. Two transmembrane proteins,
desmoglein and desmocollin, which are homologous
albeit distantly to cadherins, form the desmosomes.
Together with AJs, they provide the major intercellular
adhesive forces in polarized epithelia [5].
On the basolateral side, the membrane of a polarized cell
adheres to the extracellular matrix mainly via integrins, a
family of transmembrane proteins that induce the formation of multiprotein focal adhesion complexes [6].
The epithelium of the human gastrointestinal tract is a
typical example of an epithelial barrier held together by
TJs, AJs and desmosomes. The gastrointestinal tract
harbors up to 100 trillion commensal microorganisms that
have coevolved with the host [7] and constitutes a target
for microbial pathogens that have developed specific
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490 Celltocell contact and ECM

Figure 1

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Microbial strategies to target, cross or disrupt epithelia Sousa, Lecuit and Cossart 491

strategies to adhere to, invade or disrupt epithelial barriers [8]. These pathogens have to surmount constant
peristaltic flushing, sloughing of mucus, bile salts and
organic acids, antimicrobial peptides, defensins and competition from the resident microflora. Structural and regulatory components of the intercellular adhesion
structures, as well as matrix-binding proteins, are targets
for pathogens that interact with or cross epithelial barriers.
This review focuses on some of the striking strategies
developed by microbial pathogens to target, cross or
disrupt epithelia.

Attachment to apical epithelial surfaces and

generation of signals that disrupt junctions
A few intestinal pathogens adhere intimately to the apical
surface of epithelial cells and modify the barrier properties of the epithelium while remaining extracellular.
Enteropathogenic and enterohemorrhagic E. coli (EPEC
and EHEC) are diarrheagenic bacteria that cause the
formation of attaching and effacing (A/E) lesions, characterized by disorganization and loss of microvilli and the
generation of protruding actin-rich structures, named pedestals, at the site of bacterial attachment (Figure 1b). The
first step in the formation of an A/E lesion is the loose
attachment of the bacterium to the host epithelium via
bacterial adhesions, such as the bundle-forming pili (BFP)
[9]. Several bacterial effectors are then injected into the
host cell via a type III secretion system (TTSS). One of
these effectors, Tir, is inserted into the host cell membrane
where it adopts a hairpin loop topology. The extracellular
domain of Tir serves as receptor for intimin, an outersurface E. coli protein, and this interaction promotes tight
bacterial adhesion to the host cell [10]. Tir is the only
known example of a microbial receptor inserted by the
microbe itself in the host cell surface. The intracellular
domains of Tir (the N and C termini) recruit and activate
regulators of actin dynamics such as Nck, N-WASP and
Arp2/3 [11] and cytokeratin-18, an intermediate filament
protein (Figure 1b). The interaction of all these proteins
leads to the formation of the actin-rich pedestals characteristic of EPEC and EHEC infections [12].

EPEC affects TJs through signaling and modification of

the actin cytoskeleton. Indeed, the decrease in transepithelial resistance and appearance of aberrant TJs during
EPEC infection correlate with phosphorylation of myosin
light chain, dephosphorylation of occludin, redistribution
of ZO-1 and of occludin, and activation of ezrin
(Figure 1b) [1316]. Intimin, together with the TTSS
effectors EspF and Map, controls these effects [17]. EspF
was the first EPEC protein reported to be implicated in
the disruption of host intestinal barrier function. Map is a
multifunctional protein that carries a mitochondrial targeting sequence and promotes filopodium formation in
the early stages of EPEC infection, by an as yet unknown
mechanism [10]. Map and EspF act independently but
synergistically to alter TJs structure [17].
Clostridium difficile is responsible for pseudomembranous
colitis, a potentially severe post-antibiotic diarrhea. It
secretes two toxins, toxin A and toxin B, that bind
non-protein receptors at the host cell surface before being
endocytosed [18]. While toxin B has a potent cytotoxic
activity in vitro, the enterotoxic activity of C. difficile in
vivo is mainly attributed to toxin A [18]. Intracellularly,
both toxins monoglucosylate small GTPases (Rac, Cdc42
and Rho), inducing disorganization of the actin cytoskeleton [19]. Toxin A and toxin B also dissociate occludin,
ZO-1 and ZO-2 from cell junctions, increasing epithelium
permeability (Figure 1c) [19,20]. TJ disruption may allow
the migration of neutrophils to the site of infection, thus
creating the pseudomembranous aspect that is observed
in C. difficile-associated colitis. As both toxins glucosylate
all Rho-subfamily GTPases, they have proven to be
useful tools to evaluate the involvement of Rho GTPases
in regulatory pathways [18].
Helicobacter pylori infection is associated with chronic
gastritis, peptic ulcer, gastric adenocarcinoma and lymphoma. H. pylori expresses several adhesins that mediate
bacterial attachment to gastric epithelial glycan receptors,
and possesses a type-IV secretion system that injects the
bacterial effector CagA into the host cell cytoplasm [21].
Once inside the cell, CagA is tyrosine-phosphorylated by

(Figure 1 Legend) Schematic representation of various strategies developed by bacteria to interact with the intestinal barrier. (a) Representation
of polarized epithelial cells and components of the intercellular junctions. (b) Enteropathogenic (EPEC) adheres to the apical pole via
bundle-forming pili (BFP), resulting in attaching and effacing (A/E) lesions. Via its type-III secretion system (TTSS), it injects into the host cell
cytoplasm bacterial proteins that modify the actin cytoskeleton and interfere with tight junction (TJ) components. (c) Clostridium dificile (C.d.)
secretes two toxins (TcdA and TcdB) that interact with small GTPases and dissociate TJ proteins from the lateral membrane. (d) Helicobacter
pylori (H.p.) adheres to the epithelial apical surface and delivers CagA into the host cytoplasm. CagA interacts with TJ proteins (ZO-1 and JAM)
and activates HGF receptor, leading to the loss of TJs. Intracellular phosphorylated CagA inhibits Src kinases and activates phosphatases.
(e) Vibrio cholerae (V.c.) secretes cholera toxin (CT) that binds GM1 at the host cell surface. Intracellularly, CT stimulates the production of
cAMP, leading to water loss. V. cholerae also secretes the protease HA/P, which degrades the extracellular domain of occludin. (f) Listeria
monocytogenes (L.m.) uses two surface proteins, InlA and InlB, to enter into epithelial cells. InlA binds E-cadherin and requires catenins, vezatin
and myosin VIIa to promote entry. InlB interacts with HGF-R and stimulates the activity of PtdIns-3-kinase and cytoskeleton rearrangements.
The cell-to-cell spread of L. monocytogenes depends on the bacterial protein ActA and ezrin, a regulator of AJs assembly. (g) Bacteroides fragilis
(B.f.) secretes fragilysin, which cleaves the extracellular domain of E-cadherin, disrupting the AJs and activating b-catenin nuclear signaling.
(h) Shigella flexneri (S.f.) crosses the intestinal epithelium through M cells, using its TTSS and injecting bacterial effectors into the host cells.
Once translocated, S. flexneri enters polarized cells via the basal pole and induces the opening of Gap junctions and ATP release. The cell-to-cell
spread depends on E-cadherin and connexin 26.
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492 Celltocell contact and ECM

a Src kinase [22,23] and causes cell elongation and

increased motility. Interestingly, phosphorylated CagA
in turn inhibits Src kinase activity, whereas it stimulates
the activity of the phosphatase SHP-2, which results in
the dephosphorylation of host cell proteins such as cortactin (Figure 1d) [24]. Independently of phosphorylation
events, CagA is recruited to the host cell membrane and
interacts with ZO-1 and JAM, forming a multiprotein
complex at the bacterial attachment site and thus altering
the composition and the function of TJs [25]. In addition, CagA activates the hepatocyte growth factor receptor (HGF-R) pathway, leading to host cell scattering and
cell proliferation (Figure 1d) [26]. The activation of the
HGF-R pathway perturbs the integrity of AJs by inducing
the internalization and redistribution of the E-cadherin
molecules that make up AJs [27,28]. A plethora of effects
have been attributed to CagA, all thought to contribute to
H. pylori-associated gastro-duodenal pathologies. Interestingly, E-cadherin and IQGAP1, a regulator of AJs
formation, are translocated from the plasma membrane
to intracellular vesicles in H. pylori-infected cells [29].
Although H. pylori is classically considered as an extracellular pathogen colonizing the apical face of gastric
epithelial cells, recent studies have shown that it may
also occupy an intracellular location, in cultured cells as
well as in vivo [30,31]. The contribution of these properties to H. pylori-associated pathologies remains to be
determined.

Direct targeting of tight junction components

Extracellular bacteria may secrete or deliver bacterial
toxins that directly target the structural components of
TJs, leading to epithelial barrier dysfunction and thus
contributing to disease. Some viruses are also able to
harness TJs.
Vibrio cholerae is the causative agent of cholera, a waterborne infection characterized by watery diarrhea. It uses
its flagellum to penetrate the mucus layer, gain access to
the apical surface of enterocytes and adhere to their brush
borders. There, it produces and secretes its major toxin,
the phage-encoded cholera toxin (CT), which binds to
the host cell ganglioside GM1, its plasma membrane
receptor. CT is internalized with GM1; it then moves
to the endoplasmic reticulum via retrograde transport and
crosses its membrane [32]. In the cytoplasm, CT ADPribosylates heterotrimeric G proteins, leading to constitutive activation of adenylate cyclase and hyperproduction of cyclic AMP, which results in ion imbalance and
water excretion (Figure 1e). V. cholerae also secretes a
metalloprotease, called the hemagglutinin/protease (HA/
P), which degrades the extracellular domain of occludin
[33], an integral membrane component of TJs, thus
perturbing the barrier function of epithelial cells
(Figure 1e). The cytoplasmic domain of degraded occludin remains associated with the plasma membrane, while
ZO-1, which is not degraded by HA/P, dissociates from
Current Opinion in Cell Biology 2005, 17:489498

cellcell junctions [33]. The redistribution of ZO-1 is due

to conformational changes occurring in occludin after its
cleavage by HA/P. Note that these results are based on in
vitro studies, and that their relevance in vivo remains to be
determined. V. cholerae also produces the zonula occludens toxin (Zot), which is also phage-encoded. Zot specifically and reversibly increases the permeability of the
intestinal epithelium and is thought to be responsible for
the residual diarrhea triggered by CT mutants [34]. Its
mode of action remains elusive.
Clostridium perfringens type A is an important cause of
foodborne disease outbreaks. In the human gut, it produces the bifunctional enterotoxin CPE, which forms
pores in the host cell plasma membrane and affects TJ
structure and function [35]. The C terminus of CPE binds
claudin-3 and claudin-4 with high affinity, and induces
their degradation, a decrease in transepithelial resistance
and disruption of TJs. These findings contributed to the
demonstration that claudins play a role in the barrier
function of TJs [36]. CPE also interacts with occludin [37].
Rotavirus infection can cause severe gastroenteritis outbreaks in children. Rotavirus entry into enterocytes is a
complex multistep process involving virus surface proteins and non-structural proteins with enterotoxin activity
[38]. Rotavirus infectious particles are composed of three
concentric layers of proteins, the outer layer being mainly
built of VP4 and VP7 proteins. VP4 plays a key role in the
virushost-cell interaction and is cleaved by trypsin into
two peptides, VP5 and VP8 [38]. VP8 transiently disrupts
TJs, thus reducing transepithelial resistance, and displaces ZO-1, claudin-3 and occludin from TJs
(Figure 2a) [39]. Similar effects are produced by the viral
enterotoxin NSP4, which is released in the host cytoplasm, where it increases the intracellular calcium concentration, disrupts the actin cytoskeleton and lowers the
expression of enzymes at the apical surface (Figure 2a)
[40]. NSP4 can be exported to the extracellular medium,
where it acts on the apical surface of the enterocyte to
increase the paracellular permeability. It causes the reorganization of the actin cytoskeleton of polarized cells and
prevents lateral recruitment of ZO-1 [41]. VP7, the other
rotavirus outer surface protein, interacts with integrins
(Figure 2a) [38,42]. VP8 and NSP4 may cooperate to open
TJs in the early steps of infection, allowing the virus to
reach the basal membrane or inducing the exposure of
integrins to the apical surface.
Reoviruses are non-enveloped viruses that cause mild
gastrointestinal or respiratory diseases in children. They
use JAM-1 as a cellular receptor [43]. JAM-1 is an immunoglobulin superfamily member that forms stable homodimers and regulates TJ permeability (Figure 2a) [44].
The viral attachment protein s1 directly interacts with
JAM-1 and disrupts JAM-1 homodimers. Whether this
affects epithelial integrity is not known [45].
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Microbial strategies to target, cross or disrupt epithelia Sousa, Lecuit and Cossart 493

Figure 2

Other examples of microbial interactions with epithelia. (a) The reovirus surface protein s1 binds to JAM-1 to promote virus entry. Rotavirus
surface protein VP4 is cleaved into VP5 and VP8 proteins. VP8 displaces ZO-1, claudin-3 and occludin from tight junctions (TJs). NSP4 is a
non-structural virus protein delivered inside the host cells; it induces an increase in calcium concentration and disruption of the actin cytoskeleton.
VP7 interacts with integrins to promote entry. (b) The coxsackievirus protein Fiber interacts with CAR at the AJs during virus entry into epithelial
cells. Fiber released after virus replication binds CAR, disrupts intercellular junctions and allows the migration of new virus to the apical pole.
(c) MIC2, an adhesin of Toxoplasma gondii, interacts with ICAM-1 during its transepithelial paracellular migration. (d) The entry of human herpes
simplex virus in cells of a stratified epithelium depends on the interaction of virus glycoprotein D with nectin, a protein AJ. Human cytomegalovirus
uses two cellular receptors, EGF-R and integrin aVb3, to enter into cells. (e) The exfoliative toxin (ET) of Staphylococcus aureus (S.a.) cleaves
desmoglein1 in the epidermis, thus promoting the loss of intercellular adhesion.

Exploiting adherens junctions

Whereas the area of a polarized epithelium most accessible to luminal pathogens is its apical surface, some
pathogens target receptors located beneath TJs and have
developed strategies to reach them.
Listeria monocytogenes is a facultative intracellular pathogen with the capacity to cross the intestinal, bloodbrain
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and fetoplacental barriers. L. monocytogenes induces its

internalization into epithelial cells, mainly using two
surface proteins, InlA and InlB, which interact with host
cell receptors and mimic their physiological cellular
ligands [46]. InlA interacts specifically with human
E-cadherin and recruits b- and a-catenins, myosin VIIa
and vezatin to the site of bacterial entry (Figure 1f)
[4750]. InlA-dependent entry of L. monocytogenes has
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494 Celltocell contact and ECM

similarities to the formation and maintenance of AJs,

suggesting that the bacterium fully exploits the cellular
machinery involved in AJs formation and maintenance to
induce its own uptake. The other bacterial invasion
protein, InlB, mainly interacts with and activates the
tyrosine kinase receptor HGF-R, which colocalizes with
E-cadherin in polarized epithelial cells. The InlB/HGF-R
interaction leads to the phosphorylation of Shc, Gab1 and
Cbl and the activation of PI3-kinase, hence inducing
cytoskeleton rearrangements (Figure 1f) [51,52]. Interestingly, InlB binding to HGF-R induces HGF-R
ubiquitination, endocytosis and degradation [53]. How
L. monocytogenes gains access to the basolateral surface,
where E-cadherin is expressed, is not known. This access
may be facilitated when extruding epithelial cells detach
from the tips of intestinal microvilli, where most of the
bacteria are observed in infected intestinal tissue [54]. In
addition, the E-cadherin and HGF-R entry pathways
could act in synergy, inducing events leading to a transient depolarization of the epithelium and a possible
retargeting of E-cadherin from AJs to the apical surface.
However, the contribution of InlB in vivo at the intestinal
level appears to be insignificant (N Khelef, M Lecuit, H
Bierne and P Cossart, unpublished), arguing against this
hypothesis. Once inside the cell, L. monocytogenes is able to
spread from cell to cell: it polymerizes host actin and
moves intracellularly until it reaches the plasma membrane, forming a protrusion internalized by the neighboring cell (Figure 1f) [55]. The formation of these
protrusions depends on ezrin [56], a protein that regulates AJs assembly [57].
Bacteroides fragilis is a member of the commensal intestinal microflora that may become pathogenic when it
overgrows other bacterial species, then causing colic
abscesses and diarrhea. It secretes a zinc-dependent
metalloprotease, fragilysin, which specifically cleaves
the extracellular domain of E-cadherin (Figure 1g). This
cleavage induces the proteolysis of E-cadherin intracellular domain, leading to the disruption of Ajs and in turn
affecting TJ integrity [58]. Fragilysin also appears to be
able to trigger the activation of multiple signal transduction pathways, including b-catenin nuclear signaling [59].
Some viral species also use basolateral cell adhesion
molecules as receptors to infect epithelia. The coxsackievirus and adenovirus receptor (CAR) [60] is an adhesion
molecule belonging to the immunoglobulin superfamily
that is expressed on the basolateral membrane of polarized epithelial cells [61]. Adenovirus infects conjunctival,
respiratory, intestinal and urinary epithelia. A transient
break in epithelial integrity may allow luminal adenovirus
to reach its receptor. During the entry process, the adenovirus capsid protein fiber binds CAR, thus disrupting
CAR homodimers and leading to a decrease in epithelial
impermeability (Figure 2b) [60]. A recent study has
shown that, after infection and intracellular multiplication
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in the airway epithelium, adenoviral particles and fiber

protein are released on the basolateral cell surface. Fiber
protein interacts with high affinity with CAR, further
disrupting cellcell adhesion and allowing the virus to
travel in the intercellular space and migrate towards the
apical surface [62]. This study helped demonstrate the
role of CAR in cellcell adhesion.
Entry of herpes simplex virus (HSV) in host cells involves
the binding of HSV glycoprotein D to nectin, another
member of the immunoglobulin superfamily also present
in AJs (Figure 2d) [63,64]. Nectin is normally engaged in
homophilic interactions, although its role in the regulation of AJs is not fully understood [65,66]. While nectin is
accessible to virus in cells lacking E-cadherin, how the
virus gets access to nectin in normal epithelial cells
remains to be determined [67].
Toxoplasma gondii is a human parasite with the capacity to
actively cross all host barriers. Initially, Toxoplasma
crosses the intestinal barrier and then migrates into deeper tissues [68]. During this process, Toxoplasma parasites
accumulate around intercellular junctions and probably
use the paracellular pathway to cross epithelial barriers
without damaging their integrity. Interestingly, intercellular adhesion molecule 1 (ICAM-1), which is present in
the apical surfaces of epithelial barriers, is upregulated
during Toxoplasma infection [69]. Parasite transepithelial
migration requires the interaction of ICAM-1 with the T.
gondii surface adhesin MIC2 (Figure 2c) [70]. T. gondii
thus appears to cross epithelial barriers by following a
pathway similar to that used by transmigrating leukocytes. Interestingly, the rhinovirus, a major cause of
common cold in humans, also upregulates ICAM-1
expression and binds this protein in order to enter epithelial cells [71].

Disrupting desmosomes
Although this review mainly focuses on pathogen interactions with monolayered epithelia, such as the intestinal
epithelium, more complex epithelia such as the epidermal stratified epithelium are also targeted by human
pathogens. This is the case for Staphylococcus aureus, a
bacterium responsible for cutaneomusocal infections.
Exfoliative toxins (ETs) produced by S. aureus rapidly
diffuse into the skin tissue and cleave the cell adhesion
molecule desmoglein 1 (Dsg1), which plays a key role in
maintaining the structure and barrier function of the
epidermis (Figure 2e) [72,73]. ETs cleave a single peptide bond in the extracellular domain of Dsg1 and do not
cleave closely related molecules such as Dsg3 and
E-cadherin [74]. In addition, the specificity of ET cleavage is dependent on the calcium-stabilized structure of
Dsg1 [75]. This highly specific and efficient cleavage of
Dsg1 results in the loss of intercellular adhesion and
allows bacteria to form blisters under the epithelial sheet,
providing protective niches in which they can survive and
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Microbial strategies to target, cross or disrupt epithelia Sousa, Lecuit and Cossart 495

proliferate [76]. S. aureus can also affect the human

intestine after ingestion of food contaminated by staphylococcal enterotoxins. Staphylococcal enterotoxin B
(SEB) is an extremely potent superantigen able to activate T cells and stimulate the secretion of various cytokines. a-hemolysin is another water-soluble exotoxin
secreted by S. aureus, with a strong pore-forming activity
[77].

Other mechanisms of translocation across

epithelia
M-cells are cells of epithelial origin present within polarized epithelial cells in Peyers patches and are specialized
in the capture of intestinal luminal antigens and their
presentation to the immune system. Some pathogens,
unable to disrupt the intestinal epithelium from its apical
surface, exploit M-cells to cross the intestinal epithelium.
This is the case with Yersinia, Shigella and, to a lesser
extent, Salmonella, three bacterial species responsible for
enteroinvasive infections in humans [8].
Yersinia expresses an outer surface protein, invasin, which
binds with high affinity to b1-integrin expressed apically
by M cells. Upon invasin binding, b1-integrins form
clusters and activate signaling pathways, leading to bacterial internalization by a so-called zipper mechanism.
Once translocated across M cells, Yersinia avoids internalization into professional phagocytes by injecting into
macrophages bacterial proteins that interact with small
GTPases and thus inhibit the actin cytoskeleton rearrangements necessary for phagocytosis. In addition, it injects
the YopP/J protein, which interferes with signaling pathways, thus blocking the inflammatory response. Yersinia
multiplies extracellularly and eventually enters overlying
epithelial cells through their basal poles via an invasin
b1-integrin-mediated process [8,78]. Integrins are
exploited by several other pathogens to promote their
invasion or as receptors for toxins [79]. As recently shown,
human cytomegalovirus glycoproteins also target integrin
avb3 to invade cells [80].
Shigella enters M cells by macropinocytosis. Once translocated, Shigella causes macrophage apoptosis via the
TTSS protein IpaB, which activates caspase-1, leading
to IL-1b secretion, neutrophil recruitment, mucosal
inflammation and disruption of the epithelial barrier.
Shigella may re-enter epithelial cells via the basal surface,
using its TTSS to inject into the host cell cytoplasm the
bacterial effectors that induce membrane ruffling: IpaC
and IpaB, which interact with CD44 on the host cell
membrane, allow the injection of other bacterial effectors
that interfere with cytoskeleton machinery (Figure 1h)
[8]. Shigella cell-to-cell spreading depends on E-cadherin
and connexin 26 [81,82]. Connexins are the major
components of Gap junctions, which form intercellular
channels allowing the diffusion of small molecules
between adjacent cells (Figure 1h) [83]. Interestingly,
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Shigella invasion induces the opening of connexin-26

channels and the release of ATP to the extracellular
medium, which in turn promotes bacterial invasion and
dissemination to neighboring cells [82]. Interestingly,
proteins secreted by the human herpes simplex and
papilloma viruses are able to close the gap junctions of
infected cells, inhibiting intercellular communication
with neighboring non-infected cells [84,85]. It is possible
that this mechanism counteracts the recently reported
phenomenon that antigenic peptides may diffuse via gap
junctions and thus allow a non-infected cell to present to
the immune system microbial antigens transferred from
an adjacent infected cell [86].
Salmonella can cross the intestinal epithelium via three
different cell types: enterocytes, M cells and dendritic
cells [8]. It induces its internalization into epithelial cells
using proteins secreted by a TTSS encoded by the
Salmonella pathogenicity island 1 (SPI-1). The SPI-1encoded proteins (including SopB, SopE and SopE2)
interfere with phosphoinositides and small GTPases, thus
causing dramatic rearrangements of the actin cytoskeleton. Translocated bacteria then grow inside macrophages
using effector proteins encoded by SPI-2, a second pathogenicity island. Salmonella can also be translocated across
the intestinal barrier by dendritic cells, which thread
between enterocytes and emerge into the intestinal
lumen, where they can capture luminal antigens and
microorganisms [87].

Conclusions
Epithelia are highly organized and sophisticated structures adapted to function as barriers protecting the underlying tissues from external aggressions, including
microbial invasions. Intercellular junctions are the central
players maintaining the epithelial architecture. Apart
from being a physical obstacle, they also play a key role
in the control of cell proliferation, differentiation, migration and death. Microbial pathogens have evolved countless strategies to interfere with cellcell junctions, to
increase epithelium permeability, to destabilize epithelial structure and function and, sometimes, to cross and/or
disrupt the barrier the epithelium constitutes. This barrier is thus not invincible, and in response epithelial cells
have set up extracellular and intracellular sensing systems
(Toll-like receptors and Nods) able to detect microbial
pathogens and react to their intrusion into physiologically
sterile intracellular and extracellular environments.
Engagement of these detecting systems triggers the
recruitment and activation of effectors of the innate
and adaptive immune system at the site of infection.
In turn, these mechanisms may also be exploited by
pathogens to promote their translocation across epithelia,
and it is well known that inflammation favors secondary
infections. It is also important to recall that intestinal
epithelial cells are continuously in contact with billions of
harmless and thus so-called commensal bacteria, which
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496 Celltocell contact and ECM

they ignore (or may even recognize as friends) and

against which they do not trigger an innate or adaptive
immune response. Understanding how the host distinguishes between these two types of microbes will help us
to identify new aspects of pathogenicity and commensality. In addition, the precise elucidation of the molecular
mechanisms mediating the crossing of host barriers by
microbial pathogens represents a key issue for the comprehension of the pathophysiology of invasive infectious
diseases.

13. Muza-Moons MM, Schneeberger EE, Hecht GA:

Enteropathogenic Escherichia coli infection leads to
appearance of aberrant tight junctions strands in the lateral
membrane of intestinal epithelial cells. Cell Microbiol 2004,
6:783-793.

Acknowledgements

16. Simonovic I, Arpin M, Koutsouris A, Falk-Krzesinski HJ, Hecht G:

Enteropathogenic Escherichia coli activates ezrin, which
participates in disruption of tight junction barrier function.
Infect Immun 2001, 69:5679-5688.

The work in PCs laboratory received financial support from Institut

Pasteur (GPH N89), French Ministry of Research (Programme de
Microbiologie Fondamentale et Applique e, Maladies Infectieuses,
Environment et Bioterrorisme ACI N8 MIC0312) and Association pour
la Recherche sur le Cancer (ARC4404). S. Sousa is founded by the
Institute for Molecular and Cell Biology (POCTI-LA000308-BPD).
P. Cossart is a Howard Hughes Medical Institute International
Research Scholar.

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