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Typhoid fever: Pathogenesis and disease

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Typhoid fever: pathogenesis and disease
Deborah Housea, Anne Bishopa, Christopher Parryb, Gordon Dougana and
John Waina
Typhoid fever is an infectious disease of global distribution.
Although there is a wealth of data on Salmonella typhimurium
infection in the mouse and the interaction of this serovar with
human cell lines in vitro, there is a relatively small amount of data
on S. typhi and the pathogenesis of typhoid fever. In this review
we focus on three areas: adherence to and invasion of gut
epithelial cells, dissemination to systemic sites, and survival and
replication within host cells. In addition, we attempt to put
current salmonella research into the context of typhoid fever.
Curr Opin Infect Dis 14:573±578. # 2001 Lippincott Williams & Wilkins.
a
Centre for Molecular Microbiology and Infection, Imperial College of Science
Technology and Medicine, London, UK and bOxford University-Wellcome Trust
Clinical Research Unit, Centre for Tropical Diseases, Ho Chi Minh City, Viet Nam
Correspondence to John Wain, Centre for Molecular Microbiology and Infection,
Imperial College of Science Technology and Medicine, Exhibition Road, South
Kensington, London W7 2AZ, UK
Tel: +44 20 7594 3715; fax: +44 20 7262 6299; e-mail: j.wain@ic.ac.uk
Current Opinion in Infectious Diseases 2001, 14:573±578
Abbreviations
Sif salmonella-induced filament
SPI salmonella pathogenicity island
tcf Typhi colonization factor
TTSS type III secretion system
# 2001 Lippincott Williams & Wilkins
0951-7375
Introduction
Typhoid fever remains a major public health problem in
many regions of the world with over 16 million cases
being reported each year worldwide. Patients with
typhoid fever are infected with Salmonella enterica
subspecies I serovar Typhi (S. typhi) and usually present
to the health care worker with a history of prolonged
fever, headache, abdominal discomfort and general
lethargy. Around 10% of these develop severe or
complicated disease and without speci®c treatment 5±
30% of all patients with typhoid fever may die. Serovars
of S. enterica subspecies I (41300) cause infections in
many warm blooded animals and, despite a very close
genetic relationship to each other, display differing
ranges of host speci®city. Some serovars are host
restricted while others can infect a variety of mammals.
S. typhi, for example, is adapted, and restricted, to the
human host, whereas many isolates of S. typhimurium
cause gastroenteritis or occasionally septicaemia in
humans, lethal diarrhoea in calves or a systemic disease
in genetically susceptible mice. Since S. typhi fails to
establish infection in laboratory rodents most of the
research reported on the pathogenesis of typhoid fever is
based on in-vitro studies with human and murine cell
lines and the infection of mice with S. typhimurium. The
pathogenicity of the non-typhi Salmonella [1] and
secretion systems associated with bacterial±host cell
interactions [2] have been reviewed recently. Here we
attempt to put recent research developments into the
context of typhoid fever. We will focus on three areas:
(1) adherence to and invasion of gut epithelial cells, (2)
dissemination to systemic sites, and (3) survival and
replication within host cells.
Adherence to and invasion of epithelial cells
S. typhi is ingested in contaminated food or water. It
passes through the stomach and invades the gut
epithelium, possibly in the distal ileum. Active attach-
ment promoted by the bacterium may be necessary
before invasion can occur and this most likely involves
unidenti®ed adhesion molecules on the bacterium
interacting with receptors on the host cell. The nature
of this interaction for S. typhi is not clear but may involve
®mbriae. S. typhi has 12 ®mbrial operons of the
chaperone usher assembly class, but none, including tcf
(Typhi colonization factor) are unique to S. typhi [3 .]. S.
typhi, however, does possess a unique combination of
®mbrial operons. The diverse family of ®mbriae may be
the result of selection by the host immune response [4].
A type IVB pilus operon (pil) is also present in S. typhi
573
574 Gastrointestinal infections

and wild type (pil+) S. typhi are more able to adhere to
and invade intestinal epithelial cells than are pilS::Kmr
(pil7) mutants [5 .]. The cystic ®brosis transmembrane
conductance regulator, previously reported to be used by
S. typhi for entry into intestinal cells, has been suggested
as a possible site for attachment mediated by type IVB
pili [5 .].
avrA (a gene of unknown function) in S. typhi (Fig. 1).
Additional SPI-1 effector proteins are encoded on the
chromosome outside SPI-1. For example, in S. typhimur-
ium the gene encoding SopE2 plays an important role in
host cell invasion [8 .] and is conserved across many
pathogenic Salmonella spp. [9]. In S. typhi, however,
sopE2 is a pseudogene (Fig. 1).

Salmonella spp. invade epithelial cells in vitro by a
process of bacterial-mediated endocytosis, involving
cytoskeletal re-arrangement, disruption of the epithelial
cell brush border and the subsequent formation of
membrane ruf¯es. An adherent and invasive phenotype
of S. enterica is activated under conditions similar to
those found in the human small intestine (high
osmolarity, low oxygen). The invasive phenotype is
mediated, in part, by salmonella pathogenicity island
(SPI)-1 [6], a 40 kb region of the chromosome which
encodes regulator proteins (e.g. HilA), a type III
secretion system (TTSS) that delivers bacterial proteins
from the salmonella cytosol into the host cell, and
several effector proteins which induce changes within
the host cell and promote bacterial uptake. The
acquisition of SPI-1 probably enabled the ancestoral S.
enterica to adhere to and invade epithelial cells more
ef®ciently [7], thus allowing the colonization of a new
environmental niche. SPI-1 of S. typhimurium has been
extensively studied and a comparison of the open
reading frames of SPI-1 in S. typhimurium with those of
S. typhi reveals a single difference: the absence of the
The major regulator of SPI-1 is HilA, which is encoded
within SPI-1 itself. A number of regulators, both within
and outside SPI-1, have been identi®ed which act via
hilA [10±12]. In S. typhimurium, mutations within ¯iAZY
and ¯hCD have been reported to cause a reduction in
hilA expression [13 .], and FliZ has been independently
reported to control invasion gene expression [14 .].
Interestingly, mutations within ¯iA result in a reduction
in SPI-1 gene expression in S. typhi but not in S.
typhimurium [15 . .], suggesting that the control of
invasion genes is different in these two serovars.
The expression of invasion genes is clearly multi-
factorial and may be further complicated in S. typhi by
the presence of the polysaccharide capsule (Vi). Vi
expression is dependent upon the osmolarity of the
environment and involves the global regulator RCS-AB
[16]. Under conditions of low osmolarity, Vi is down-
regulated, which enhances the secretion of SPI-1
effector proteins and promotes the expression of an
adherent and invasive phenotype. Indeed, deletion of
the Vi biosynthetic genes enhances the secretion of SPI-

Figure 1. A schematic representation showing the organization of genes within the SPI-1 region of Salmonella typhimurium and the nature of
the proteins that they encode

SPI-1 genes in S.typhimurium

sit avr spr hil org prg hil iag spt sic iac sip sic spa inv
A B C D A B C C B A K J I H D A B P P P A D C B A S R Q P O N M C B A E G F H

Effectors outside of SPI-1 that are

secreted by the SPI-1 TTSS

sop sop sop sop sop hsp

D E E2 B A H1

X X X

S. typhimurium genes that have been reported not to be present, or to be pseudogenes, in S. typhi are marked with a cross. Some of these genes have
alternative names: sipA-D = sspA-D; sopB = sigD; hilA = iagA; hilC = sirC = sprA. , Secreted effectors and translocon components; , Iron
uptake system; , Transcription regulators; , Type III secretory apparatus; , Chaperones involved in type III secretion; , Unknown
function.

1 products and allows expression of an adherent and
invasive phenotype of S. typhi at any osmolarity [17].
A number of SPI-1 effector proteins have been
described. Some of these, such as SipB and SipC,
associate with the plasma membrane of epithelial cells
and may facilitate penetration of eukaryotic cell mem-
branes by the SPI-1 TTSS [18,19 . .], while others affect
the actin cytoskeleton arrangement within the host cell.
The SopB effector protein has broad speci®city inositol
phosphatase activity in vitro and contributes, along with
uncharacterized SopE-dependent signals, to an increase
in inositol-(1,4,5,6)-tetraphosphate levels which result in
actin reorganization within the infected host cell [20].
SopB is also required for activation of Akt/PKB, a kinase
that induces anti-apoptotic survival signals [21,22]. All of
these genes are present in S. typhi but their role in
typhoid fever needs to be con®rmed.
Although SPI-1 is important for the invasion of epithelial
cells in vitro, the role of this island for invasion in the
different steps of infection in vivo in less clear [23].
Several workers report that SPI-1 knock out Salmonella
spp. are less able to colonize animal hosts [24] but S.
typhimurium in which SPI-1 is deleted (D-spi1::kan)
colonize susceptible mice as well as wild-type S.
typhimurium in either mixed or single oral infections
[25 .]. Thus, it would appear that there are alternative
mechanisms by which Salmonella may cross the epithe-
lial border.
Dissemination of Salmonella typhi to
systemic sites via phagocytic cells
Although S. typhimurium can cause systemic disease in
the human host [26], infection with this serovar usually
results in a localized enteritis associated with a secretory
response in the intestinal epithelium and recruitment
and transmigration of neutrophils into the intestinal
lumen [27,28]. S. typhimurium induces human intestinal
epithelial cells to secrete interleukin-8 [29 .] and other
pathogen-elicited epithelial chemoattractants [30 .] which
direct the recruitment and transmigration of neutrophils
into the gut lumen (Fig. 2a). S. typhi is not typically
associated with acute diarrhoea, suggesting that the initial
interaction between this serovar and the human gut is less
in¯ammatory than that seen with enteritis-causing
Salmonella spp. The lack of acute in¯ammation and the
absence of subsequent recruitment of neutrophils may
allow S. typhi to invade into the deeper tissues of the gut,
although there is little experimental evidence for this
hypothesis. S. typhi does, however, stimulate IL-6
secretion from human epithelial cells [31]. The magni-
tude of the response is dependent upon the isolate of S.
typhi (clinical versus laboratory {Ty2}) and is higher than
with S. typhimurium, possibly re¯ecting differences in
adherence/invasion, which is greater for S. typhi [32]. A
Typhoid fever: pathogenesis and disease House et al. 575
Figure 2. Comparison of the events occurring within the gut lumen
and gut-associated tissues which may determine whether an
infection with Salmonella will be limited to the gut (a) or become
systemic (b)
(a)
(4) Gut lumen
(1)
PEEC
(2)
(3)
IL-8
IL-8
(b) (1) Gut lumen
(4)
(2)
IL-6
(3)
IL-1 β
(a) (1) Exposure to the environment within the gut lumen upregulates the
expression of SPI-1 and induces the release of flagellin. (2) Flagellin is
translocated across the epithelium and is released onto the basolateral
surface, where it binds to an unidentified receptor and induces
interleukin (IL)-8 secretion. (3) IL-8 recruits neutrophils to the site of
infection. (4) SipB activates protein kinase C (PKC) and induces
secretion of pathogen-elicited epithelial chemoattractants (PEECs) at
the apical surface of the epithelial cell. PEECs direct the transmigration
of the newly-recruited neutrophils into the gut lumen. (b) (1) Exposure to
the environment within the gut lumen upregulates the expression of SPI-
1 and induces the release of flagellin. Vi expression is downregulated,
which further promotes the secretion of SPI-1 effector proteins and the
release of flagellin. (2) The invasive Salmonella traverse the gut
epithelium. Interaction with and invasion of epithelial cells with
Salmonella induce the secretion of IL-6. The invading bacteria are taken
up by/invade macrophages within the gut-associated tissue. (3) SPI-1-
expressing Salmonella induce caspase-1-dependent rapid death of the
infected macrophages, resulting in the release of IL-1. The release of
proinflammatory cytokines recruits blood monocytes and other inflam-
matory cells to the site of infection. (4) The invading bacteria are
disseminated throughout the body. This may be within CD18+ cells that
have been recruited to the site of infection.
recent in-vitro study used high density cDNA microarrays
to investigate the epithelial cell response to infection with
S. dublin and this approach may prove useful for the
investigation of differential host response to the different
Salmonella serovars [33 . .].
Shortly following invasion of the gut epithelium,
invasive Salmonella spp. encounter macrophages within
the gut-associated lymphoid tissue. The interaction
between Salmonella and macrophage results in an
alteration in the expression of a number of host genes
including those encoding pro-in¯ammatory mediators
576 Gastrointestinal infections
(e.g. iNOS, chemokines, interleukin-1b), receptors or
adhesion molecules (e.g. TNFaR, CD40, ICAM-1), and
anti-in¯ammatory mediators (e.g. TGFb1 and 2) [34 . .].
Other up-regulated genes include those involved in cell
death or apoptosis (e.g. ICE protease, TNFR1, Fas) and
transcription factors (e.g. Egr-1, IRF-1). Some non-typhi
serovars can induce rapid macrophage death (within
30 min of infection) in vitro [35]. This is mediated by the
SPI-1 effector SipB, is dependent upon the host cell
protein caspase-1 and has features common to both
apoptosis and necrosis [36,37]. Caspase-1 mediated cell
death is a pro-in¯ammatory process [38 .] that would
seem to be counter-productive if an organism is to
establish a systemic infection. However, this response
appears to be necessary in the murine model as S.
typhimurium cannot establish a systemic infection in
caspase-17/7 mice [39]. The pro-in¯ammatory response
and the subsequent recruitment of phagocytic cells to
the site of the infection may facilitate systemic spread of
the bacteria [23]. Salmonella-induced caspase-1 mediated
cytotoxicity can occur in association with, or indepen-
dently of, the activation of the initiator caspase, caspase-
2 [40]. The cytotoxic potential of S. typhi has not been
fully evaluated, although it appears to be lower than with
S. typhimurium, both in murine and human monocytic
cell lines [41]. Fig. 2b summarizes the sequence of
events that might occur within the gut tissue and which
result in the establishment of a systemic infection by
invasive Salmonella spp.
Survival, replication and transmission
of Salmonella typhi
It is generally accepted that an ability to survive within
monocytes/macrophages is essential for S. typhimurium to
establish a systemic infection in the mouse. In the
absence of SPI-1 expression, Salmonella-infected macro-
phages can remain viable for several hours [38 .] and thus
serve as a cellular niche in which the bacteria can survive
protected from the host immune system. Intracellular S.
typhimurium are located within specialized Salmonella-
containing vacuoles that have diverged from the normal
endocytic pathway. Within epithelial cells, lysosomal
glycoprotein (lgb)-containing tubular structures, termed
Salmonella-induced ®laments (Sifs), can be seen radiating
from Salmonella-containing vacuoles. Although Sifs are
not seen in S. typhimuriun-infected macrophages, sifA7
S. typhimurium are unable to survive well and replicate
ef®ciently within these cells in vitro and are attenuated
in vivo when administered intraperitoneally to mice
[44 . .,45]. Data from mixed infections suggest that SifA is
secreted via the SPI-2 TTSS [44 . .] and limited
homology between SifA and other putative SPI-2
secreted proteins has been described [46].
A high proportion of S. typhi in the bone marrow of the
human host is intracellular [42] and in the blood this
percentage increases as the disease progresses [43]. It is
possible, therefore, that S. typhi may traf®c around the
host inside cells, although this has not been shown
de®nitively. Studies on the invasion properties of S. typhi
in humans have been limited. Vaccine strains of S. typhi
harbouring mutations in aro genes can be detected in the
blood of humans following an oral challenge [47]. Similar
strains of S. typhi (Ty2) with deletions in aroC and ssaV, a
component of the SPI-2 TTSS, however, do not cause
vaccinaemia (D. Lewis, personal communication), sug-
gesting that SPI-2 may be required for S. typhi to
establish a systemic infection in humans. Shedding of S.
typhi in the faeces of an infected individual is an essential
step in the transmission of typhoid fever. There are very
few data on the genetic basis of shedding. Although a
role for the shdA gene in the transmission of S.
typhimurium has been demonstrated [48], the closest
match for this gene in the S. typhi CT18 genome appears
to be a pseudogene.
Conclusion
This year the sequencing of both the S. typhi (CT18) and
S. typhimurium (LT2) genomes was completed (see
http://www.sanger.ac.uk and http://genome.wustl.edu).
Although 95% of genes are shared (98% identity)
between S. typhi and S. typhimurium a detailed compar-
ison of the remainder of the genomes will aid in the
identi®cation of genes contributing to host speci®city
and serovar speci®c disease. These comparisons are
already under way and one of the most striking ®ndings
so far is the presence of single mutations or deletions
within the genome of S. typhi, which appear to generate
many pseudogenes, while the corresponding open read-
ing frames in S. typhimurium are intact. The inactivation
of single genes, as well as the acquisition or loss of single
genes or large islands of DNA, may have contributed to
host adaptation and restriction of S. typhi.
Although it is dif®cult to study humans infected with S.
typhi, either naturally or experimentally, these studies are
invaluable. A recent study demonstrated an association
between susceptibility to typhoid fever and genes within
the major histocompatability complex class II and class
III loci (TNFA*2{-308}´DRB1*0301) [49 . .]. This is
similar to the situation in murine enteric fever when
susceptibility to infection is dependent upon major
histocompatability complex class II antigens. No associa-
tion was found, however, between susceptibility to
typhoid fever and natural resistance associated macro-
phage protein 1 [50], a locus associated absolutely with
susceptibility to Salmonella infections in the mouse.
There is a wealth of data on S. typhimurium infection in
the mouse and the interaction of this serovar with human
cell lines in vitro, but there are relatively few data on S.
typhi and the pathogenesis of typhoid fever. If we are to
gain a greater understanding of the disease process in

typhoid fever, experiments comparing the interaction of
S. typhi, S. typhimurium and other S. enterica serovars with
human cells and tissues are required.
References and recommended reading
Papers of particular interest, published within the annual period of review, have
been highlighted as:
. of special interest
.. of outstanding interest
1 Groisman EA, Mouslim C. Molecular mechanisms of salmonella pathogen-
esis. Curr Opin Infect Dis 2000; 13:519±522.
2 Plano GV, Day JB, Ferracci F. Type III export: new uses for an old pathway.
Mol Microbiol 2001; 40:284±293.
3 Townsend SM, Kramer NE, Edwards R, et al. Salmonella enterica serovar
. Typhi possesses a unique repertoire of fimbrial gene sequences. Infect Immun
2001; 69:2894±2901.
This paper describes a sequence analysis of S. typhi for fimbrial operons followed
by a salmonella-wide survey for their distribution. Although no clear relationship
between fimbrial operons and host specificity was shown, S. typhi was found to
have a unique set of fimbrial antigens.
4 Nicholson TL, Baumler AJ. Salmonella enterica serotype Typhimurium elicits
cross-immunity against a Salmonella enterica serotype enteritidis strain
expressing LP fimbriae from the lac promoter. Infect Immun 2001; 69:204±
212.
5 Zhang XL, Tsui IS, Yip CM, et al. Salmonella enterica serovar Typhi uses type
. IVB pili to enter human intestinal epithelial cells. Infect Immun 2000; 68:3067±
3073.
Using INT407 cells, pilS mutants of S. typhi were shown to be deficient in cell
invasion. Adhesion of wild-type S. typhi was prevented by addition of PilS to the
medium but adhesion of S. typhimurium was not affected. Two conclusions were
drawn: (1) the type IVB pili of S. typhi may be involved in adhesion to intestinal
cells; and (2) S. typhi and S. typhimurium adhere to INT407 cells in culture by
different mechanisms.
6 Sukhan A. The invasion-associated type III secretion system of Salmonella
typhimurium: common and unique features. Cell Mol Life Sci 2000; 57:1033±
1049.
7 Kingsley RA, Baumler AJ. Host adaptation and the emergence of infectious
disease: the Salmonella paradigm. Mol Microbiol 2000; 36:1006±1014.
8 Stender S, Friebel A, Linder S, et al. Identification of SopE2 from Salmonella
. typhimurium, a conserved guanine nucleotide exchange factor for Cdc42 of the
host cell. Mol Microbiol 2000; 36:1206±1221.
This paper describes the cloning of S. typhimurium sopE2 and the ability of SopE2
to activate Cdc42. It also observes that S. typhi sopE2 is probably a pseudogene.
9 Bakshi CS, Singh VP, Wood MW, et al. Identification of SopE2, a Salmonella
secreted protein which is highly homologous to SopE and involved in
bacterial invasion of epithelial cells. J Bacteriol 2000; 182:2341±2344.
10 Altier C, Suyemoto M, Ruiz AI, et al. Characterization of two novel regulatory
genes affecting Salmonella invasion gene expression. Mol Microbiol 2000;
35:635±646.
11 Fahlen TF, Mathur N, Jones BD. Identification and characterization of mutants
with increased expression of hilA, the invasion gene transcriptional activator
of Salmonella typhimurium. FEMS Immunol Med Microbiol 2000; 28:25±35.
12 Wilson RL, Libby SJ, Freet AM, et al. Fis, a DNA nucleoid-associated protein,
is involved in Salmonella typhimurium SPI-1 invasion gene expression. Mol
Microbiol 2001; 39:79±88.
13 Lucas RL, Lostroh CP, DiRusso CC, et al. Multiple factors independently
. regulate hilA and invasion gene expression in Salmonella enterica serovar
Typhimurium. J Bacteriol 2000; 182:1872±1882.
This paper gives a good summary of the gene products reported to influence
invasion gene expression. It reports on a S. typhimurium screen for hilA expression
deficient mutants that reveals multiple independent regulatory systems.
14 Iyoda S, Kamidoi T, Hirose K, et al. A flagellar gene fliZ regulates the expression
. of invasion genes and virulence phenotype in Salmonella enterica serovar
Typhimurium. Microb Pathog 2001; 30:81±90.
Identification of FliZ as a regulator of hilA expression in S. typhimurium that may
co-ordinate motility and the expression of invasion genes.
Typhoid fever: pathogenesis and disease House et al. 577
15 Eichelberg K, Galan JE. The flagellar sigma factor FliA (sigma(28)) regulates the
. . expression of Salmonella genes associated with the centisome 63 type III
secretion system. Infect Immun 2000; 68:2735±2743.
This paper reports on the effects of loss of function mutations in genes which
regulate flagella. Mutations in flhDC or fliA dramatically reduced macrophage
toxicity and entry of S. typhi into cultured epithelial cells but this effect was much
reduced with S. typhimurium. The reduced invasiveness of the S. typhi filA mutant
could not be restored by centrifugation and was associated with reduced SPI-1
gene expression. The authors conclude that there is an overlap between the
regulatory mechanisms of flagella and type III secretion systems in S. typhi.
16 Wehland M, Bernhard F. The RcsAB box. Characterization of a new operator
essential for the regulation of exopolysaccharide biosynthesis in enteric
bacteria. J Biol Chem 2000; 275:7013±7020.
17 Zhao L, Ezak T, Li ZY, et al. Vi-suppressed wild strain Salmonella typhi
cultured in high osmolarity is hyperinvasive toward epithelial cells and
destructive of Peyer's patches. Microbiol Immunol 2001; 45:149±158.
18 Scherer CA, Cooper E, Miller SI. The Salmonella type III secretion translocon
protein SspC is inserted into the epithelial cell plasma membrane upon
infection. Mol Microbiol 2000; 37:1133±1345.
19 Hayward RD, McGhie EJ, Koronakis V. Membrane fusion activity of purified
. . SipB, a Salmonella surface protein essential for mammalian cell invasion. Mol
Microbiol 2000; 37:727±739.
Paper observing that SipB, a S. typhimurium translocon component, associates
with epithelial cell membranes and has membrane binding and membrane fusion
activities in vitro, implying that SipB may facilitate the insertion of the SPI-1 TTSS
into eukaryotic cell membranes.
20 Zhou D, Chen LM, Hernandez L, et al. A Salmonella inositol polyphosphatase
acts in conjunction with other bacterial effectors to promote host cell actin
cytoskeleton rearrangements and bacterial internalization. Mol Microbiol
2001; 39:248±259.
21 Steele-Mortimer O, Knodler LA, Marcus SL, et al. Activation of Akt/protein
kinase B in epithelial cells by the Salmonella typhimurium effector sigD. J Biol
Chem 2000; 275:37718±37724.
22 Marcus SL, Wenk MR, Steele-Mortimer O, et al. A synaptojanin-homologous
region of Salmonella typhimurium SigD is essential for inositol phosphatase
activity and Akt activation. FEBS Lett 2001; 494:201±207.
23 Vazquez-Torres A, Jones-Carson J, Baumler AJ, et al. Extraintestinal
dissemination of Salmonella by CD18-expressing phagocytes. Nature
1999; 401:804±808.
24 Wallis TS, Wood M, Watson P, et al. Sips, Sops, and SPIs but not Stn
influence Salmonella enteropathogenesis. Adv Exp Med Biol 1999; 473:275±
280.
25 Murray RA, Lee CA. Invasion genes are not required for Salmonella enterica
. serovar Typhimurium to breach the intestinal epithelium: evidence that
salmonella pathogenicity island 1 has alternative functions during infection.
Infect Immun 2000; 68:5050±5055.
This study uses a competitive index to investigate the role of hilA and SPI-1 in the
establishment of a systemic infection of S. typhimurium in the mouse following an
oral challenge. Loss of function of hilA, but not deletion of SPI-1, resulted in
reduced colonization of systemic organs compared to wild-type.
26 Graham SM, Molyneux EM, Walsh AL, et al. Nontyphoidal Salmonella
infections of children in tropical Africa. Pediatr Infect Dis J 2000; 19:1189±
1196.
27 Fleckenstein JM, Kopecko DJ. Breaching the mucosal barrier by stealth: an
emerging pathogenic mechanism for enteroadherent bacterial pathogens. J
Clin Invest 2001; 107:27±30.
28 Ohl ME, Miller SI. Salmonella: a model for bacterial pathogenesis. Annu Rev
Med 2001; 52:259±274.
29 Gewirtz AT, Simon Jr PO, Schmitt CK, et al. Salmonella typhimurium
. translocates flagellin across intestinal epithelia, inducing a proinflammatory
response. J Clin Invest 2001; 107:99±109.
The secretion of interleukin-8 by epithelial cells in response to infection is well
documented. This study provides further information on the antigens that can
induce this response. They show that the flagellin antigen is translocated across
epithelial cell monolayers where it then interacts with an as yet unidentified
receptor to induce interleukin-8 secretion. Interleukin-8 secretion was not
observed in S. typhimurium in which phoP was constitutively active.
 578 Gastrointestinal infections
30 Lee CA, Silva M, Siber AM, et al. A secreted Salmonella protein induces a
. proinflammatory response in epithelial cells, which promotes neutrophil
migration. Proc Natl Acad Sci U S A 2000; 97:12283±12288.
This study shows that the SPI-1 protein SipA alone is sufficient to elicit neutrophil
transmigration across human polarized epithelial cells. SipA itself is not a
chemoattractant but instead appears to interact with the apical surface of the
epithelial cells, resulting in the activation of PKC and the subsequent trans-
epithelial migration of neutrophils. This demonstrates that some SPI-1 effector
proteins involved in invasion of epithelial cells may have additional roles that do not
require their introduction directly into the cytosol of the host cell.
31 Weinstein DL, O'Neill BL, Metcalf ES. Salmonella typhi stimulation of human
intestinal epithelial cells induces secretion of epithelial cell-derived interleukin-
6. Infect Immun 1997; 65:395±404.
32 Weinstein DL, O'Neill BL, Hone DM, et al. Differential early interactions
between Salmonella enterica serovar Typhi and two other pathogenic
Salmonella serovars with intestinal epithelial cells. Infect Immun 1998;
66:2310±2318.
33 Eckmann L, Smith JR, Housley MP, et al. Analysis by high density cDNA arrays
. . of altered gene expression in human intestinal epithelial cells in response to
infection with the invasive enteric bacteria Salmonella. J Biol Chem 2000;
275:14084±14094.
See ref. [34..].
34 Rosenberger CM, Scott MG, Gold MR, et al. Salmonella typhimurium infection
. . and lipopolysaccharide stimulation induce similar changes in macrophage gene
expression. J Immunol 2000; 164:5894±5904.
These studies (Refs [33..] and [34..]) used high-density cDNA micro-arrays to
investigate the host cell response (epithelial or macrophage) to infection with non-
typhoidal Salmonella. This global approach to the investigation of gene expression
shows that a variety of genes are up-regulated in response to an infection. These
include cytokines and cytokine receptors, adhesion and cell surface molecules,
and proteins associated with the cell cycle and cell death. In the study using
murine macrophages, the host response to S. typhimurium was very similar to that
seen with LPS, suggesting that, under certain circumstances, the immediate host
cell response to this pathogen can be dominated by the somatic antigen. This type
of experimental approach should prove useful in comparisons of different
Salmonella serovars or mutants.
35 Navarre WW, Zychlinsky A. Pathogen-induced apoptosis of macrophages: a
common end for different pathogenic strategies. Cell Microbiol 2000; 2:265±
273.
36 Brennan MA, Cookson BT. Salmonella induces macrophage death by
caspase-1-dependent necrosis. Mol Microbiol 2000; 38:31±40.
37 Watson PR, Gautier AV, Paulin SM, et al. Salmonella enterica serovars
Typhimurium and Dublin can lyse macrophages by a mechanism distinct from
apoptosis. Infect Immun 2000; 68:3744±3747.
38 Boise LH, Collins CM. Salmonella-induced cell death: apoptosis, necrosis or
. programmed cell death? Trends Microbiol 2001; 9:64±67.
This concise review summarizes many of the recent studies of Salmonella induced
apoptosis and cell death.
View publication stats
39 Monack DM, Hersh D, Ghori N, et al. Salmonella exploits caspase-1 to
colonize Peyer's patches in a murine typhoid model. J Exp Med 2000;
192:249±258.
40 Jesenberger V, Procyk KJ, Yuan J, et al. Salmonella-induced caspase-2
activation in macrophages: a novel mechanism in pathogen-mediated
apoptosis. J Exp Med 2000; 192:1035±1046.
41 Schwan WR, Huang XZ, Hu L, et al. Differential bacterial survival, replication,
and apoptosis-inducing ability of salmonella serovars within human and
murine macrophages. Infect Immun 2000; 68:1005±1013.
42 Wain J, Bay PV, Vinh H, et al. Quantitation of bacteria in bone marrow from
patients with typhoid fever: relationship between counts and clinical features.
J Clin Microbiol 2001; 39:1571±1576.
43 Wain J, Diep TS, Ho VA, et al. Quantitation of bacteria in blood of typhoid
fever patients and relationship between counts and clinical features,
transmissibility, and antibiotic resistance. J Clin Microbiol 1998; 36:1683±
1687.
44 Beuzon CR, Meresse S, Unsworth KE, et al. Salmonella maintains the integrity
. . of its intracellular vacuole through the action of SifA. EMBO J 2000; 19:3235±
3249.
This study provides evidence of a role for SifA in the survival of S. typhimurium
within macrophages. SifA appears to localize to the cytosolic face of the
salmonella-containing vacuole where it recruits host membranes and thus allows
the vacuole to increase in size around the replicating bacteria. In-vivo studies of
mixed infections suggest that the expression of SifA is regulated by the SPI-2
regulator SsaV and that the protein is secreted via the SPI-2 TTSS.
45 Brumell JH, Rosenberger CM, Gotto GT, et al. SifA permits survival and
replication of Salmonella typhimurium in murine macrophages. Cell Microbiol
2001; 3:75±84.
46 Miao EA, Miller SI. A conserved amino acid sequence directing intracellular
type III secretion by Salmonella typhimurium. Proc Natl Acad Sci U S A 2000;
97:7539±7544.
47 Tacket CO, Hone DM, Curtiss RD, et al. Comparison of the safety and
immunogenicity of delta aroC delta aroD and delta cya delta crp Salmonella
typhi strains in adult volunteers. Infect Immun 1992; 60:536±541.
48 Kingsley RA, van Amsterdam K, Kramer N, et al. The shdA gene is restricted
to serotypes of Salmonella enterica subspecies I and contributes to efficient
and prolonged fecal shedding. Infect Immun 2000; 68:2720±2727.
49 Dunstan SJ, Stephens HA, Blackwell JM, et al. Genes of the class II and class III
. . major histocompatibility complex are associated with typhoid fever in Vietnam. J
Infect Dis 2001; 183:261±268.
This is one of the first studies to document an association between the host's
genetic background and susceptibility to human typhoid fever. The authors
demonstrate an association between genes within the MHC class II and class III
region (HLA-DR, HLA-DQ, and TNFA) on chromosome 6 and either resistance or
susceptibility to typhoid fever. The study utilized two unrelated sample sets from
geographically distinct locations in southern Vietnam.
50 Dunstan SJ, Ho VA, Duc CM, et al. Typhoid fever and genetic polymorphisms
at the natural resistance-associated macrophage protein 1. J Infect Dis 2001;
183:1156±1160.