Future

Editorial

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Medicinal
Chemistry

Retrofitting the battlements: tight junction

remodeling as a novel antimicrobial
approach
...serious attention should be paid to the molecular modification of epithelial
defenses...
Keywords: claudin Ebola epithelial barrier function hepatitis HIV tight junction
transepithelial virus

Epithelial cell layers, pathogens

&infection
Imagine that you are tasked with defending a
castle from an invading army. You have two
general options: you can attack the invaders directly or you can redefine your castles
walls. It would probably be wise to exercise
both options, and yet modern medicines
approach to microbial pathogens has generally utilized only the first, a varied yet monolithic drive to kill the microbes coming over
or through the walls. Prescribers of antimicrobial agents know quite well that this
approach can exert a high toll on your own
troops. In addition, the enemy is quite good
at adapting its defense and tactics to changes
you make to your armory, as demonstrated
by the current epidemic of antibiotic-resistant organisms. In this article, we wish to
bring attention to recently gained knowledge
in the world of epithelial tissue biology and
specifically epithelial tight junction (TJ)
complexes, which may open up a new range
of options to stopping microbial pathogens at
the epithelial gates.
Except for the unusual venue of contracting
an infection through an open wound or transcutaneous penetration, such as intravenous
drug use, there is an amazing universality
regarding a microbes modus operandi a virus,
bacterium or fungus must first compromise
and penetrate an epithelial cell layer/barrier
in order to cause systemic illness. The first tissue of ours that these microbes encounter is
an epithelial barrier: epidermis, nasopharyngeal mucosa, bronchial lining, gastrointestinal mucosa, cornea, urethra or vaginal lining,

10.4155/FMC.14.146 2015 Future Science Ltd

among others. In 99% of cases, the very first

task of the microbe is to latch onto this barrier
and traverse it.
Higher animal life with humanity being
no different is basically a parallel array of
sacs and tubes. The sac or tube is always an
epithelial tissue. From a microbes perspective, on the luminal side lies the untamed
environment, with its less than optimal (and
highly variable) living conditions. On the
abluminal side is the vasculature with its
promise of well-regulated conditions and an
uninterrupted food supply. It is no surprise
that pathogens evolved with the ability to
transverse these epithelial barriers and enter
the abluminal space.
Key interplay of pathogens
&epithelial TJs
An interesting convergence of broad concepts
is occurring at this time in the TJ/barrier
integrity research field. On the one hand,
there is an emerging realization of how a
broad range of pathogens both viral and
bacterial are fixated on TJs. This has been
very excellently reviewed several times in the
last 10years [1,2] . This fixation occurs in two
modalities (Figure 1) . First, certain pathogens actually target junctional complexes as
docking sites on the epithelial barrier, binding to one or more TJ proteins and using this
binding to facilitate invasion of either the
epithelial cell itself or the paracellular space.
The bacterium best known for this action
is Clostridium perfringens, whose secreted
toxin (CPE) likely binds more than one TJ
protein [35] . On the viral level, hepatitis Cs

Future Med. Chem. (2015) 7(1), 913

James M Mullin
Author for correspondence:
The Lankenau Institute for Medical
Research, 100 Lancaster Avenue,
Wynnewood, PA 19096, USA
Division of Gastroenterology, Lankenau
Medical Center, Wynnewood,
PA19096,USA
mullinj@mlhs.org

Jonathan Raines
The Lankenau Institute for Medical
Research, 100 Lancaster Avenue,
Wynnewood, PA 19096, USA

LawrenceLLivornese Jr
The Division of Infectious Diseases,
Lankenau Medical Center, Wynnewood,
PA 19096, USA

part of

ISSN 1756-8919

Editorial Mullin, Raines & Livornese

(HepC) viral envelope proteins interact with the TJ
protein claudin-1 in almost a receptor-like relationship
in order to facilitate viral entry into the cell, with possible involvement of occludin and claudin-6 and -9 as
well [6] . Second, a broader class of pathogens again
both viral and bacterial induce disruption of the TJ
complex as a result of their infection/invasion of the
epithelial cell, an eventuality that can not only lead to
pronounced morbidity (in part, but not merely, due to
induction of proinflammatory cascades arising out of
luminal antigen entry into the abluminal space), but
also to vastly increased pathogen entry from the luminal compartment into the abluminal space following
the same paracellular leak pathways. This second phenomenon applies to a broad list of microbes, among
them being HepC, enteropathogenic Escherichia coli,
Helicobacter pylori, adenovirus, HIV and respiratory
syncitial virus [1,711] . In point of fact, certain pathogens
may be able to pursue both options, with C. perfringens
and HepC being likely examples [1,12] .

That such modification can be achieved by

micronutrients seems to be a gift thrown our way by
our own evolution.

Micronutrient leverage on epithelial TJs

A second recent emerging realization in biomedical
research is that an increasing and diverse array of naturally occurring and synthetically derived compounds
exist that can induce remodeling of TJs, leading to a
less leaky junctional complex (and consequently a less
leaky barrier). The group of naturally occurring micronutrient compounds with this capability is quite diverse
[13] . This phenomenon has similarly been reviewed by
several research groups in the last 10years [14,15] . These
compounds can be as simple as the trace metal zinc
[16] and the commensal bacterial metabolite indole [17]
or the relatively complex shRNAs that target specific
regulators of TJ components [18] . This enhancement of
TJ barriers by phosphorylation-based and/or transcriptionally based remodeling of the complex is no mean
feat, given that the universal effect of the host of pathophysiological and pathogenic components of disease in
general is to increase TJ leak. This is true whether we
are talking about cancer [19] , stress [20,21] , inflammation
[22] or the microbial-based maladies mentioned above.
However, in the published biomedical literature, we
have not as yet seen the connection made that a novel
means of reducing the efficiency of the microbial infection of epithelial cells and their barriers may be to prophylactically induce TJ remodeling, if not actual TJ
enhancement. On the molecular scale of things, pathogens tend to be relatively large entities, and it is quite
possible that their binding (e.g., to claudin-3) may be

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Future Med. Chem. (2015) 7(1)

highly influenced by the immediate transmembrane

molecular neighbors of claudin-3 (e.g., other claudins
and occludin, among others), and that changing this
molecular neighborhood may significantly affect the
efficiency with which a given pathogen can interface
with the TJ complex. The dietary and other agents
mentioned above do just this by altering the relative
proportions of the various TJ proteins to each other in
the TJ complex. It should be kept in mind that victory in this overall effort lies not in preventing binding, but merely making it less efficient, and thereby
reducing infection efficiency and the infectious load
to be presented to the immune system. Similarly, for
the pathogens that merely induce TJ leakiness but do
not necessarily bind to TJs, it may be interesting to see
whether some of these newly recognized compounds
that can render TJs less susceptible to induced leakiness from proinflammatory stimuli (they could also be
used prophylactically to reduce the TJ leakiness arising
from pathogen infection of epithelial cells; Figure 1B)
[23,24] . The result would be to reduce pathogen presentation to the basallateral cell surface (where many
pathogen receptors are localized) and also to stem the
influx of pathogens to the interstitial fluid compartment and vasculature. As mentioned above, this could
not only reduce morbidity during certain microbial
infections, but more importantly could reduce the
bacterial or viral load crossing a barrier and presenting to the immune system. This could be the difference between an infection remaining subclinical or
progressing to septic shock.
The measles example

It is instructive to look at the case of measles virus

(MV) infection of epithelia as an example of pathogen infection of epithelia that one would not initially
associate with TJs or TJ modulators. First, MV infection of epithelia appears to be primarily mediated
by MV contact with the basallateral cell surface
[25,26] . This is true even though the virus appears to
be shed apically from infected cells [26] . So, either initial (environmental) presentation of MV to epithelial
cells or infection of epithelia from virus shed by other
(infected) epithelia has an intrinsic problem: it comes
at the epithelium from the wrong surface (in this case
the apical surface). The fact that disrupted TJs may
significantly enhance the MV infection of an epithelial
layer was dramatically shown by Shirogane et al.,who
simultaneously demonstrated that disruption could
not proceed so far as to induce dedifferentiation of
the cell (i.e., MV has a distinct preference for a differentiated [polar] epithelial cell layer, particularly one
with impaired TJs) [27] . This impairment of TJs could
theoretically ensue from MV infection itself or, more

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Retrofitting the battlements: tight junction remodeling as a novel antimicrobial approach

Lumen

Paracellular space

Tight junction
complex

Lumen

Particle virus
permeation into
Junctional
the paracellular
compromise
space is reduced
by healthy tight
junction complexes

Tight junction
complex

Interstitium

Interstitium

Lumen

Editorial

Virus binding to
the tight junction
strands is
decreased in
the remodeled
junctions

Lumen

Viruses enter
cells at
junctions
after binding
Micronutrient to specific
treatment
claudins or
occludins

Interstitium

Interstitium

Figure 1. Two different scenarios wherein the state of a tight junction complex can significantly influence viral invasion across an
epithelial barrier. (A) Pathogen entry into the paracellular space (then interstitial fluid and vasculature) can be facilitated by an
event (humoral or directly caused by the pathogen itself) that induces tight junctional leakiness. Access into the paracellular space
allows contact with basallateral cell membrane receptor molecules, which is necessary for pathogen entry into the epithelial cell,
as well as subsequently unobstructed entry into the interstitial fluid space and vasculature. For example, this would be part of HIVs
modus operandi. (B) Certain pathogens are able to bind directly to tight junctional proteins, thereby facilitating their entry into the
epithelial cell in internalized complexes of virus and junctional proteins. For example, this would be an etiology used by hepatitis C
virus.

typically, from some other unrelated, serendipitous

morbidity that renders leakythe TJs of the targeted
epithelium beforehand. Malnutrition is one such predisposing serendipitous condition that is capable of

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inducing epithelial barrier leak [28,29] . Induced barrier

leak could well be one explanation for why malnutrition associates with poor outcomes in infections such
as measles. However, if one prophylactically brings

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Editorial Mullin, Raines & Livornese

into play an agent or agents that are capable of modifying the TJs and rendering them less susceptible to
compromise (e.g., the micronutrients shown to offset
TJ and barrier compromise by proinflammatory cytokines), then one might quantitatively and materially
affect the ability of MV to infect/invade an epithelial
target tissue. In this light, it is very interesting to see
that vitamin A has been shown to positively affect the
clinical course of MV infection [30] . It is tempting to
assume that the documented enhancement of TJ barrier function (and TJ protein remodeling) by vitamin
A [31,32] may play a role here.

Financial & competing interests disclosure

Conclusion
In opposing an invading horde, it is advisable to
improve the battlements in addition to attacking the
enemy.With this in mind, serious attention should be
paid to the molecular modification of epithelial defenses

The authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment,
consultancies, honoraria, stock ownership or options, expert
testimony, grants or patents received or pending, or royalties.
No writing assistance was utilized in the production of this
manuscript.

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