The structures of Escherichia coli O-polysaccharide antigens
Roland Stenutz1, Andrej Weintraub2 & Göran Widmalm1
1
Department of Organic Chemistry, Arrhenius Laboratory, Stockholm University; and 2Karolinska Institutet, Department of Laboratory Medicine,
Division of Clinical Bacteriology, Karolinska University Hospital, Stockholm, Sweden
Correspondence: Andrej Weintraub,
Karolinska Institutet, Department of
Laboratory Medicine, Division of Clinical
Bacteriology, F82, Karolinska University
Hospital, Huddinge, S-14186 Stockholm,
Sweden. Tel.: 146 858 587831; fax: 146 871
13918; e-mail: [email protected]
Received 31 August 2005; revised 15 November
2005; accepted 21 November 2005.
First published online 9 February 2006.
doi:10.1111/j.1574-6976.2006.00016.x
Editor: Simon Cutting
Keywords
Enterobacteriacea, serotype, O-antigen,
structure, NMR, database.
Abstract
Escherichia coli is usually a non-pathogenic member of the human colonic flora.
However, certain strains have acquired virulence factors and may cause a variety of
infections in humans and in animals. There are three clinical syndromes caused by
E. coli: (i) sepsis/meningitis; (ii) urinary tract infection and (iii) diarrhoea.
Furthermore the E. coli causing diarrhoea is divided into different ‘pathotypes’
depending on the type of disease, i.e. (i) enterotoxigenic; (ii) enteropathogenic;
(iii) enteroinvasive; (iv) enterohaemorrhagic; (v) enteroaggregative and (vi)
diffusely adherent. The serotyping of E. coli based on the somatic (O), flagellar
(H) and capsular polysaccharide antigens (K) is used in epidemiology. The
different antigens may be unique for a particular serogroup or antigenic
determinants may be shared, resulting in cross-reactions with other serogroups of
E. coli or even with other members of the family Enterobacteriacea. To establish
the uniqueness of a particular serogroup or to identify the presence of common
epitopes, a database of the structures of O-antigenic polysaccharides has
been created. The E. coli database (ECODAB) contains structures, nuclear
magnetic resonance chemical shifts and to some extent cross-reactivity relationships. All fields are searchable. A ranking is produced based on similarity,
which facilitates rapid identification of strains that are difficult to serotype
(if known) based on classical agglutinating methods. In addition, results pertinent
to the biosynthesis of the repeating units of O-antigens are discussed. The
ECODAB is accessible to the scientific community at http://www.casper.organ.
su.se/ECODAB/.
Introduction
Escherichia coli is the type species of the genus Escherichia
that contains mostly motile Gram-negative bacilli that fall
within the family Enterobacteriaceae. It is the predominant
facultative anaerobe of the human colonic flora. The organism typically colonizes the infant gastro-intestinal tract
within hours after birth, and E. coli and the host derive
mutual benefit for the rest of the host’s life (Kaper et al.,
2004). However, several E. coli clones have acquired specific
virulence factors which increase their ability to adapt to new
niches and allow them to cause a broad spectrum of diseases.
Three general clinical syndromes can result from infection
with pathogenic E. coli strains: enteric/diarrhoeal disease;
urinary tract infection; and sepsis/meningitis (Nataro &
Kaper, 1998). As long as these bacteria do not acquire
genetic elements encoding for virulence factors, they remain
benign commensals. Strains that acquire bacteriophage or
2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
c
plasmid DNA encoding enterotoxins or invasion factors
become virulent. Among the E. coli causing intestinal
diseases, there are six well-described pathotypes: enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC),
enteroinvasive E. coli (EIEC), enterohaemorrhagic E. coli
(EHEC), enteroaggregative E. coli (EAEC) and diffusely
adherent E. coli (DAEC) (Nataro & Kaper, 1998). These
pathotypes have virulence attributes that help bacteria to
cause diseases by different mechanisms.
Enteric/diarrhoeal Escherichia coli
Enteropathogenic Escherichia coli (EPEC)
Enteropathogenic Escherichia coli was the first pathotype of
Escherichia coli to be described. Large outbreaks of infant
diarrhoea in UK led Bray, in 1945, to describe a group of
serologically distinct E. coli strains that were isolated from
FEMS Microbiol Rev 30 (2006) 382–403
383
Escherichia coli O-polysaccharide antigens
children with diarrhoea but not from healthy children
(Kaper et al., 2004). The hallmark of infections due to EPEC
is the attaching-and-effacing histopathology, which can be
observed in intestinal biopsy specimens from patients or
infected animals (Nataro & Kaper, 1998). The most prevalent serogroups within this group of E. coli are: O18ac,
O20, O25, O26, O44, O55, O86, O91, O111, O114, O119,
O125ac, O126, O127, O128, O142 and O158 (Nataro &
Kaper, 1998).
Enteropathogenic Escherichia coli infection is primarily a
disease of infants younger than 2 years (Nataro & Kaper,
1998). EPEC primarily causes acute diarrhoea, although
many cases of persistent EPEC diarrhoea have been reported
(Nataro & Kaper, 1998; Scaletsky et al., 1996). In addition to
watery diarrhoea, vomiting and low-grade fever are common symptoms of EPEC infection. EPEC plays a more
important role in developing countries where it is the
foremost cause of diarrhoea. Many case-control studies have
found EPEC to be more frequently isolated from children
with diarrhoea than from the controls. Studies in Brazil,
Mexico, and South Africa have shown that 30–40% of infant
diarrhoea can be attributed to EPEC (Robins-Browne et al.,
1980; Cravioto et al., 1988, 1990; Gomes et al., 1989, 1991).
Recently, the pathogenesis of EPEC has been reviewed from
the historical point of view and although the pathotype has
been described in the 1940s, the exact mechanism of the
disease is not completely understood (Chen & Frankel,
2005).
Enterotoxigenic Escherichia coli (ETEC)
Enterotoxigenic Escherichia coli is a common cause of
infectious diarrhoea (Black, 1993), especially in tropical
climates, where uncontaminated water is not readily available. Most of the illnesses, in terms of both numbers of cases
and severity of symptoms, occur in infants and young
children after weaning. This pathogen may express heatlabile and/or heat-stable toxins. Heat-labiles are a class of
enterotoxins that are closely related in structure and function to cholera enterotoxin, which is expressed by Vibrio
cholerae O1 and O139 (Sixma et al., 1993). The genes
encoding heat-labile and heat-stable toxins are carried on
plasmids. ETEC colonizes the surface of the small bowel
mucosa and elaborates enterotoxins, which give rise to
intestinal secretion. Colonization is mediated by one or
more proteinaceous fimbrial or fimbrillar adhesins termed
colonization factor antigens (CFA) (Kaper et al., 2004). A
single plasmid often carries a toxin and CFA, for example,
heat-stable toxin and CFA/I (Reis et al., 1980; McConnell
et al., 1981; Murray et al., 1983), heat-labile and heat-stable
toxins and CFA/II (Penaranda et al., 1983; Smith et al.,
1983), and heat-stable toxin and CFA/IV (Thomas et al.,
1987). The clinical features of ETEC diarrhoea are consistent
FEMS Microbiol Rev 30 (2006) 382–403
with the pathogenic mechanism of ETEC enterotoxins.
ETEC diarrhoea may be mild, brief, and self-limiting or
may be as severe as that seen in V. cholerae infection (Levine
et al., 1977; Wolf, 1997). The percentage of ETEC in children
with diarrhoea varies from 10% to 30% (Albert et al., 1992;
Mangia et al., 1993; Hoque et al., 1994; Flores Abuxapqui
et al., 1999). Several studies suggest that 20–60% of travellers
from developed countries experience diarrhoea when visiting the areas where ETEC infection is endemic; 20–40% of
the cases are due to ETEC (Black, 1990; Arduino & DuPont,
1993; DuPont & Ericsson, 1993). The most common ETEC
serogroups are: O6, O8, O11, O15, O20, O25, O27, O78,
O128, O148, O149, O159 and O173.
Enteroinvasive Escherichia coli (EIEC)
Enteroinvasive Escherichia coli is a pathogenic form of E. coli
that can cause dysentery (Nataro & Kaper, 1998). EIEC
strains are biochemically, genetically and pathogenically
closely related to Shigella spp. The precise pathogenic
scheme of EIEC has yet to be elucidated. However, pathogenesis studies of EIEC suggest that its pathogenic features
are virtually identical to those of Shigella spp. (Goldberg &
Sansonetti, 1993; Parsot & Sansonetti, 1996). Genes necessary for invasiveness are carried on a 120-MDa plasmid in
Shigella sonnei and a 140-MDa plasmid in other Shigella
species and in EIEC (Baudry et al., 1987; Small & Falkow,
1988; Sasakawa et al., 1992). EIEC penetrates the intestinal
mucosa, predominantly that lining the large intestine, to
cause inflammation and mucosal ulceration that are characteristic of bacillary dysentery.
The most severe manifestation of infection with Shigella
spp. and EIEC is bacillary dysentery, a syndrome characterized by frequent small-volume stools with blood and mucus.
The disease is responsible for a substantial proportion of
acute diarrhoeal diseases worldwide. However, most persons
infected with Shigella spp. or EIEC experience watery
diarrhoea that may or may not be followed by dysentery
(Snyder et al., 1984; Nataro et al., 1998; Taylor et al., 1988).
In most cases, EIEC elicits watery diarrhoea that is indistinguishable from that caused by other E. coli pathotypes
(Nataro et al., 1998). EIEC can cause outbreaks of gastroenteritis. In sporadic cases, EIEC may be misidentified as
Shigella spp. or non-pathogenic E. coli strains. EIEC outbreaks are usually food-borne or waterborne (Nataro et al.,
1998). The most common EIEC serogroups are: O28ac,
O29, O112ac, O124, O136, O143, O144, O152, O159, O164
and O167.
Enterohaemorrhagic Escherichia coli (EHEC)
Enterohaemorrhagic Escherichia coli is an etiological agent
of diarrhoea with life-threatening complications. EHEC
belongs to a group of E. coli called VTEC (‘verotoxigenic
2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
c
384
E. coli’ or ‘Vero cytotoxin-producing E. coli’) or STEC
(‘Shiga toxin-producing E. coli’), formerly SLTEC (‘Shigalike toxin producing E. coli’). It is believed that this
pathotype adheres to the colon and distal small intestine;
however, typical lesions have not been demonstrated (Kehl,
2002). The best-characterized adherence phenotype is the
intimate or attaching and effacing adherence mediated by
the eaeA gene. STEC isolates that possess the eaeA gene are
capable of producing diarrhoea. However, the pathological
lesions associated with haemorrhagic colitis and haemorrhagic uremic syndrome are due to the action of Shiga toxin
(Stx) with endothelial cells. The term ‘enterohaemorrhagic
E. coli’ (EHEC) was originally coined to denote strains that
cause haemorrhagic colitis and haemorrhagic uremic syndrome, express Stx, cause attaching-and-effacing lesions on
epithelial cells, and possess a c. 60-MDa plasmid (Levine &
Edelman, 1984; Levine, 1987). Thus, EHEC denotes a subset
of STEC. Whereas not all STEC strains are believed to be
pathogens, all EHEC strains by the above definition are
considered to be pathogens. EHEC can cause nonbloody
diarrhoea, bloody diarrhoea, and haemorrhagic uremic
syndrome in all age groups, but the young and the elderly
are the most susceptible. The most notorious E. coli serotype
associated with EHEC is O157:H7, which has been the cause
of several large outbreaks of disease in North America,
Europe and Japan (Boyce et al., 1995; Grimm et al., 1995;
Kaper, 1998; Ozeki et al., 2003; Ezawa et al., 2004). The most
common EHEC serogroups are: O4, O5, O16, O26, O46,
O48, O55, O91, O98, O111ab, O113, O117, O118, O119,
O125, O126, O128, O145, O157 and O172. Recently, several
new EHEC serogroups have been described: O176, O177,
O178, O179, O180 and O181 (Scheutz et al., 2004). In
addition, many of the EHEC serogroups are also identified
as EPEC.
Enteroaggregative Escherichia coli (EAEC)
Enteroaggregative Escherichia coli is defined as E. coli that do
not secrete heat-labile or heat-stable enterotoxins and adhere to HEp-2 cells in an aggregative pattern (Nataro &
Kaper, 1998; Nataro et al., 1998). The basic strategy of EAEC
seems to comprise colonization of the intestinal mucosa,
probably predominantly that of the colon, followed by
secretion of enterotoxins and cytotoxins (Nataro et al.,
1998). Studies on human intestinal specimens indicate that
EAEC induces mild, but significant, mucosal damage (Hicks
et al., 1996). The clinical features of EAEC diarrhoea are
increasingly well defined in outbreaks, sporadic cases and
the volunteer model. A growing number of studies have
supported the association of EAEC with diarrhoea in developing countries, most prominently in association with
persistent diarrhoea (Bhan et al., 1989a–c; Fang et al., 1995;
Lima et al., 1992). Previous studies in children less than 5
2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
c
R. Stenutz et al.
years of age, all with diarrhoea or acute diarrhoea, have
shown a significant difference in the EAEC prevalence
compared to the controls (Nataro et al., 1987; Cravioto
et al., 1991; Bhatnagar et al., 1993; Bouzari et al., 1994;
Gonzalez et al., 1997). The increasing number of such
reports and the rising proportion of diarrhoeal cases in
which EAEC is implicated suggest that this pathotype is an
important emerging agent of paediatric diarrhoea. The
serogroups that have been identified within the EAEC group
are O3, O7, O15, O44, O77, O86, O111, O126 and O127.
Diffusely adherent Escherichia coli (DAEC)
Diffusely adherent Escherichia coli is a category of E. coli that
produces a diffuse adherence in the HEp-2 cell assay (Nataro
et al., 1998). Little is known about the pathogenesis of
DAEC. A surface of fimbria that mediates diffuse adherence
phenotype has been cloned and characterized (Bilge et al.,
1993a, b, 1989; Kerneis et al., 1991). The gene encoding the
fimbria can be found on either the bacterial chromosome or
a plasmid. Few epidemiological and clinical studies have
been carried out to be able to describe adequately the
epidemiology and clinical aspect of diarrhoea caused by
DAEC. In one study, the patients with DAEC had watery
diarrhoea without blood and faecal leukocytes (Poitrineau
et al., 1995). The association of DAEC with diarrhoea has
been shown in some studies (Giron et al., 1991; Jallat et al.,
1993; Levine et al., 1993) but not in others (Gunzburg et al.,
1993; Germani et al., 1996; Scaletsky et al., 2002).
Urinary tract infections
Uropathogenic Escherichia coli (UPEC)
The urinary tract is among the most common sites of
bacterial infection and Escherichia coli is by far the most
common infecting agent at this site. The subset of E. coli that
causes uncomplicated cystitis and acute pyelonephritis is
distinct from the commensal E. coli strains that make up
most of the E. coli populating the lower colon of humans.
E. coli from a small number of O serogroups – O4, O6, O14,
O22, O75 and O83 – cause 75% of these urinary tract
infections. Furthermore, they have phenotypes that are
epidemiologically associated with cystitis and acute pyelonephritis in the normal urinary tract. Clonal groups and
epidemic strains that are associated with urinary tract
infections have been identified (Phillips et al., 1988; Manges
et al., 2001). Although many urinary tract infection isolates
seem to be clonal, there is no single phenotypic profile that
causes urinary tract infections. Specific adhesins, including
P (Pap), type 1 and other fimbriae, seem to aid in colonization (Phillips et al., 1988; Nowicki et al., 1989; Johnson,
1991; Manges et al., 2001).
FEMS Microbiol Rev 30 (2006) 382–403
385
Escherichia coli O-polysaccharide antigens
Sepsis/meningitis
Meningitis/sepsis associated Escherichia coli
(MNEC)
This Echerichia coli pathotype is the most common cause of
Gram-negative neonatal meningitis, with a case fatality rate
of 15–40% and severe neurological defects in many of the
survivors (Unhanand et al., 1993; Dawson et al., 1999). A
majority (80%) of the E. coli strains that cause meningitis
possess the K1 capsular polysaccharide.
Other potential Escherichia coli pathotypes
Several other potential E. coli pathotypes have been described, but none of these is as well established as the
pathotypes described above. Among the most intriguing of
these potential pathogens are strains of E. coli that are
associated with Crohn’s disease and are known as adherentinvasive E. coli (Darfeuille-Michaud, 2002). An inflammatory process and necrosis of the intestinal epithelium are
characteristics of necrotizing enterocolitis (NEC), an important cause of mortality and long-term morbidity in preterm infants. Necrotoxic E. coli (NTEC) have been associated with disease in both humans and animals (De Rycke
et al., 1999). The relationships among the NEC-associated
strains, NTEC and strains associated with Crohn’s disease
have not yet been clearly established. A poorly characterized
subset of E. coli infections outside the gastrointestinal or
urinary tract is a group implicated in intra-abdominal
infections, including abscesses, wounds, appendicitis and
peritonitis.
Typing of Escherichia coli
There have been several available assays to identify different
categories of diarrhoeagenic Escherichia coli. Isolation and
identification of E. coli based on the biochemical properties
are widely used in most microbiological laboratories as they
do not require sophisticated equipment or complicated
protocols. E. coli can be easily recovered from clinical
samples on general or selective media at 37 1C under aerobic
conditions. E. coli are usually identified by biochemical
reactions. In general, the different pathotypes cannot be
identified based on biochemical criteria alone, as in most
cases they are indistinguishable from non-pathogenic E. coli.
In addition to the biochemical tests, serology is commonly used. It is based on Kauffmann’s scheme for the
serologic classification of E. coli, which is extensively
reviewed in (Orskov & Orskov, 1984; Ewing, 1986). Serotyping E. coli is performed on the basis of their O (somatic),
H (flagellar), and K (capsular) surface antigen profile. More
than 180 O, 60 H, and 80 K antigens have been proposed
(Whitfield & Roberts, 1999; Robins-Browne & Hartland,
FEMS Microbiol Rev 30 (2006) 382–403
2002). Each O antigen defines a serogroup. E. coli of specific
serogroups can be associated with certain clinical syndromes
(Nataro & Kaper, 1998; Campos et al., 2004). A specific
combination of O and H antigens defines the ‘serotype’ of
an isolate. One pathotype can comprise several serogroups
and one serogroup may belong to several pathotypes and
even to non-pathogenic E. coli (Nataro & Kaper, 1998;
Campos et al., 2004). Due to the limited sensitivity and
specificity, and the various combinations of antigens, serotyping is tedious and expensive and is performed reliably
only by a small number of reference laboratories.
Among the most useful methods to diagnose different
pathotypes of E. coli are phenotypic assays, which are based
on the virulence characteristics. Of them, the HEp-2 adherence assay is useful to identify the adherence patterns of
diarrhoeagenic E. coli. It remains the ‘gold standard’ for the
diagnosis of EAEC and DAEC (Vial et al., 1990; Nataro
et al., 1998; Donnenberg & Nataro, 1995). Identification of
ETEC has relied on the detection of heat-labile and/or heatstable enterotoxins. The classical phenotypic assay for EIEC
identification is the Sereny (guinea pig keratoconjunctivitis)
test, which correlates with the ability of the strain to invade
epithelial cells and spread from cell to cell (Kopecko, 1994).
Molecular genetic methods remain the most popular and
most reliable techniques for differentiating pathogenic
strains from non-pathogenic members. The assays are based
on nucleic acid probes and PCR and have been extensively
used. The advantages of PCR include its high sensitivity in
detection of target templates and both rapid and reliable
results due to its high specificity (Schultsz et al., 1994;
Ramotar et al., 1995; Stacy-Phipps et al., 1995; Kai et al.,
2000; Dutta et al., 2001; Pulz et al., 2003; Gioffre et al.,
2004).
Shigellae
Shigellae are Gram-negative, non-motile, facultative anaerobic rods. Shigella are differentiated from the closely related
E. coli on the basis of pathogenicity, physiology (failure to
ferment lactose or decarboxylate lysine) and serology (Samuel, 1996). The genus is divided into four species with
multiple serotypes: Shigella dysenteriae (12 serotypes), Shigella flexneri (6 serotypes), Shigella boydii (18 serotypes) and
S. sonnei (1 serotype) (Samuel, 1996). Shigella enterotoxin 1
(ShET1) is found in S. flexneri 2a, but it is only occasionally
found in other serotypes. In contrast, ShET2 is more widespread and detectable in 80% of Shigella representing all
four species. Shigella dysenteriae serotype 1 expresses Shiga
toxin, an extremely potent, ricin-like cytotoxin that inhibits
protein synthesis in susceptible mammalian cells. This toxin
also has enterotoxic activity in rabbit ileal loops, but its
role in human diarrhoea is unclear. Shiga toxin is associated
with haemorrhagic uremic syndrome, a complication of
2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
c
386
infections with S. dysenteriae serotype 1. Closely related
toxins are expressed by EHEC strains including the potentially lethal, food-borne O157:H7 serotype (Samuel, 1996).
The four Shigella species cause varying degrees of dysentery, characterized by fever, abdominal cramps and diarrhoea containing blood and mucous. Shigellosis is endemic
in developing countries where sanitation is poor. In developed countries, single-source, food or water-borne outbreaks occur sporadically, and pockets of endemic
shigellosis can be found in institutions and in remote areas
with substandard sanitary facilities. Isolation and identification of Shigella spp. is usually based on culture, biochemical
tests, and serotyping. Molecular methods can be used to
determine some target genes.
Lipopolysaccharide
Lipopolysaccharide (LPS), also known as endotoxin, is
anchored in the outer membrane of the Gram-negative
bacterium. It consists of three parts: lipid A, which is the
toxic component; the core region, which can be divided into
an inner and an outer part; and finally the O-antigen
polysaccharide, which is specific for each serogroup (Fig. 1)
(Brade et al., 1999). The sugar residues in lipid A and the
core region are decorated to a varying extent with phosphate
groups or phosphodiester-linked derivatives, which ensures
microheterogeneity in each strain. The lipid A part is highly
conserved in Escherichia coli. The core, however, contains
five different basic structures, denoted R1 to R4 and K12.
The O-polysaccharide is linked to a sugar in the outer core.
The O-antigen usually consists of 10–25 repeating units
containing two to seven sugar residues. Thus, the molecular
mass of the LPS present in smooth strains will be up to
25 kDa.
The present scheme of E. coli O-antigens comprises O1 to
O181. The following O groups have been removed: O31,
O47, O67, O72, O93, O94 and O122. The O93 strain,
however, will probably be re-introduced (Scheutz et al.,
2004). Escherichia coli strain 73-1 has been typed as E. coli
O73:K :H33 and strain 62D1 was suggested to belong to
the genus Erwinia herbicola (Scheutz, 2004). In several cases
the O-antigens of E. coli are identical or nearly identical to
those of other bacteria (Table 1).
Fig. 1. Schematic structure of an enterobacterial lipopolysaccharide
molecule. The lipids are depicted by curved lines and the sugar residues
are as follows: GlcN (’), Kdo (.), heptose (m), hexose (^), and Oantigen components (), most commonly hexose.
2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
c
R. Stenutz et al.
Structural determination of O-antigens
from strains that are difficult to type or of
nontyped strains
The serotyping of clinical isolates of Escherichia coli is under
constant development and usually it is possible to identify
the isolated strains. In some cases, however, it is not possible
properly to characterize the strain with available monospecific polyclonal antisera, either due to auto agglutination or
because the isolated E. coli strain is novel and appropriate
antisera have not been raised. Under such circumstances it is
of great interest to have a procedure that rapidly could
indicate, independently of immunological tests, the serotype
of the isolated strain.
Since immunochemical tests require cultivation of the
strain, we obtain sufficient material for analysis by other
methods.
Nuclear magnetic resonance spectroscopy is a powerful
tool that is used for studying biomolecules, including
bacterial polysaccharides. In structural studies of these
polysaccharides, NMR signals from the polymers may be
observed from live bacteria preparations or of the extracted
LPS. In the structural determination of the O-antigen
polysaccharide part of an LPS, the O-polysaccharide is often
released from the lipid A part by treatment with dilute acid
and purified by gel permeation chromatography. These steps
are laborious and should be omitted, in particular, if only
typing of the strain is required.
As it is easy to perform the phenol-water extraction from
the cultivated bacterial isolates to obtain LPS, we focus on a
procedure that rapidly can identify the most probable Oantigenic formula from a crude LPS preparation. A 1H NMR
spectrum of the LPS in D2O can be obtained in a few
minutes. Such a spectrum contains a number of characteristic signals, even though most of them are not resolved (Fig.
2). To utilize the information contained in a 1H NMR
spectrum, a database with the structures of the E. coli Oantigen polysaccharides was implemented. Each structure
has published NMR data associated with it as well as
described cross-reactivity when present. Links to the original publications are also provided in the web-based implementation. In the following approach we often enter sugar
components of the O-polysaccharide as some or all can be
determined in a few hours by chemical derivatization and
analysis with gas-liquid chromatography/mass spectrometry
(GLC-MS), high performance liquid chromatography
(HPLC) or electrophoresis techniques from a hydrolysate of
the polymer, where the choice of technique for practical
reasons is the one used in each investigator’s laboratory.
We will now exemplify the approach by analysis of E. coli
isolates that were not possible to serotype. Two clinical
isolates of E. coli from children with diarrhoea in León,
Nicaragua, termed strains 97RN and 121RN, showed
FEMS Microbiol Rev 30 (2006) 382–403
387
Escherichia coli O-polysaccharide antigens
Table 1. Escherichia coli O-antigens identical or nearly identical to
other bacterial polysaccharides
Serogroup
Identical to
Ref.
O8
Klebsiella pneumoniae O5
Serratia marcescens S3255
Hafnia alvei PCM 1223
Jansson et al. (1985)
Aucken & Pitt (1991)
Katzenellenbogen et al.
(2001)
Prehm et al. (1976)
Oxley & Wilkinson (1986)
Staaf et al. (1999a)
Rundlöf et al. (1998)
Kenne et al. (1983b)
Dmitriev et al. (1977)
Perry & MacLean (1987)
Marsden et al. (1994)
O9
O18
O21
O35
O55
O58
O97
O98
O104
O105
O111
O121
O124
O143
O147
O157
Klebsiella pneumoniae O3
Serratia marcescens O8
Hafnia alvei O39
Salmonella enterica O62
Salmonella enterica O50
Shigella dysenteriae type 5
Yersinia enterocolitica O5,27
Yersinia enterocolitica
O11,24
Escherichia coli K9
Shigella boydii type 11
Salmonella enterica O:35
Shigella dysenteriae type 7
Shigella dysenteriae type 3
Shigella boydii type 8
Shigella flexneri type 6
Citrobacter sedlakii NRCC
46070
Citrobacter freundii F90
Citrobacter freundii OCU158
Salmonella enterica O30
Gamian et al. (1992)
L’vov et al. (1991)
Kenne et al. (1983b)
Parolis et al. (1997)
Dmitriev et al. (1976)
Landersjö et al. (1996)
Hygge Blackeman et al.
(1998)
Vinogradov et al. (2000)
Bettelheim et al. (1993)
Nishiuchi et al. (2000,
2002)
Bundle et al. (1986)
identical 1H NMR spectra (cf. Fig. 2) and contained glucose,
galactose and glucosamine according to GLC analysis. These
sugar components together with selected 1H NMR data were
entered to the web-based search interface, which then selects
a best fit to the records in the database. The results of this
search gave a close match to the O-antigen structure of
E. coli O21 (and E. coli strain 105). Further inspection and
comparison of NMR data confirmed the identity between
Fig. 2. 1H nuclear magnetic resonance spectrum of the lipopolysaccharide from Escherichia coli strain 97RN in D2O solution.
FEMS Microbiol Rev 30 (2006) 382–403
the strains. Thus, the procedure rapidly revealed the serogroup of these two strains and no further structural
investigation was necessary.
Biosynthesis considerations
The biosynthesis of an LPS molecule and its transport to the
outer membrane of Gram-negative bacteria depend on
several complex events taking place at different locations in
the bacterium (Raetz & Whitfield, 2002; Samuel & Reeves,
2003). For the synthesis of the O-chain part, two of the three
reported pathways are present in Escherichia coli, namely, the
Wzy-polymerase-dependent pathway present in most cases
and typical for heteropolysaccharides and the ABC-transporter-dependent pathway, typical for homopolymers. Once
the nucleotide sugars have been synthesized they can be
incorporated into the growing O-chain. In the Wzy-dependent pathway a glycosyl-1-phosphoryl residue is transferred
to an undecaprenyl phosphate acceptor to form an undecaprenyl-PP-sugar intermediate. Subsequent transfer of additional sugars to this acceptor results in an undecaprenyl-PPoligosaccharide intermediate in which the sequence of
sugars is related to the biological repeating unit to be formed
in the O-chain. Translocation of this intermediate occurs
from the cytoplasmic side of the membrane to the periplasmic side in a Wzx-dependent process. The Wzy-dependent
polymerization of the O-antigen occurs at the reducing end
of the nascent chain being formed, meaning that the Ochain on the undecaprenol-PP carrier is transferred to the
most recently synthesized undecaprenol-PP-oligosaccharide. The extent of polymerization, i.e. the chain-length
modality, is determined by the Wzz product. The action of
the Wzy-polymerase from a linear undecaprenol-PP-oligosaccharide to produce a branched structure with a sidechain offers several possibilities just at this step to produce
different structures with regard to anomeric configuration,
linkage position and sugar residue.
The ABC-transporter pathway utilizes the b-D-GlcNAcPP-undecaprenol entity as a primer for the chain elongation
taking place on the cytoplasmic side of the membrane. In E.
coli O9, a homopolymer of mannose, an adaptor (a-D-Man)
is (1 ! 3)-linked to the N-acetylglucosamine residue. Subsequent chain growth occurs by processive glycosyl transfer
to the non-reducing terminus. In E. coli O8, also a homopolymer of mannose, the O-chain is terminated by a 3-Omethyl-D-Man residue. Although the sugars are added one
by one, sodium dodecyl sulphate-polyacrylamide electrophoresis (SDS-PAGE) analysis of these LPS molecules reveal
distributions of distinct bands. It is therefore reasonable to
describe, also in this case, the repeating units of the O-chain
in the context of biological repeating units. The undecaprenyl-linked polymer depends on Wzm and Wzt for transfer
to the periplasmic face of the membrane. For both these
2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
c
388
pathways the O-chain-PP-undecaprenyl entity is ligated to
the Lipid A-core acceptor and subsequently translated to the
outer membrane.
In Shigella flexneri the O-antigens have different structures
as a result of acquisition of genetic material from bacteriophages via transduction (Lerouge & Vanderleyden, 2001).
The glucosyl residues, present as side-chains in the repeating
unit, are proposed to be transferred to the growing
O-antigen chain on the periplasmic side of the membrane.
A similar pathway could be possible for some of the E. coli
O-antigens, as indicated by their substituent sugars and their
location within the repeating unit of the polymer (vide infra).
We also note that a gene has been identified for a
glucosylphosphate transferase, which then is responsible for
the formation of the phosphodiester-linked glycosyl residue
within the repeating unit of the O-antigen of E. coli O172
(Guo et al., 2004). Thus, this finding indicates that the
‘phospho-sugar’ is transferred en bloc in the biosynthesis.
O-antigen repeating units: characteristics
and statistics of the structures
In humans, only a handful of different sugar residues are
utilized in most glycoconjugates such as glycolipids and
glycoproteins (Varki et al., 1999). In bacteria, however, a
large number of different sugars are found and the Oantigens of Escherichia coli contain a great variety of them
(Table 2). In addition, a number of unusual sugars are found
in these polymers (Scheme 1), including pentoses, deoxyhexoses, lactyl substituted hexoses, heptoses and nonuloses.
The number of sugar residues in the O-antigen repeating
unit ranges from two to seven and the topology of the
repeats may be described as linear, branched or double
branched. We have analysed the topology based on the
number of sugar residues in the backbone (Table 3). By far,
the most common topology contains four sugars in the
backbone being linear or containing a single terminal
residue in the side-chain. The 3- and 5-residue backbones
are also common, whereas the 2- and 6-residue backbones
are only present in a few cases.
Each sugar residue is found in either the a- or the bconfiguration at the anomeric centre. The common sugars
(including ring form) of E. coli O-antigens, viz., D-Glcp, DGlcpNAc, D-Galp, D-GalpNAc, D-Manp, and L-Rhap are all
found with both anomeric configurations. Other sugars, e.g.
L-FucpNAc or D-Quip4NAc, have hitherto only been found
in one of the anomeric configurations, namely the a- or the
b-configuration, respectively. Some of the sugars in the sidechains are present only as nonterminal residues, e.g. DGlcpNAc, whereas others are only found at a terminal
position in the biological repeating unit, e.g. Colp. The
unusual groups are then highly accessible and consequently
specific for that particular E. coli serogroup.
2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
c
R. Stenutz et al.
Table 2. Abundance of glycosyl residues in Escherichia coli O-antigens
Anomers
Sidechains
Sugar
a1b
a
b
Anyw
Terminalz
Internal‰
Colp
L-Fucp
L-FucpNAc
D-Glcp
D-GlcpA
D-GlcpNAc
D-Galf
D-Galp
D-GalpA
D-GalpNAc
D-GalpNAcA
D-Manp
Neu5Ac
D-Quip3NAc
D-Quip4NAc
L-Rhap
D-Ribf
L-6dTalp
Other k
1
3
2
11
2
19
2
12
2
13
1
10
1
1
1
12
1
1
5
1
3
2
7
o1
7
o1
6
1
8
1
6
1
o1
0
10
0
1
3
0z
o 1z
0
3
2
13
2
7
1
5
0
4
0
1
1
2
1
0
2
6
10
13
13
0
8
32
2
8
2
18
25
4
0
4
13
38
0
15
0
23
4
0
2
12
2
2
4
21
4
4
8
4
0
0
4
In percent of 361 residues.
w
Abundance in any sidechain. Percent of 50 residues.
Abundance in sidechains in which the glycosyl group is directly linked to
a branch-point residue, which itself is adjacent to and substituted by a
2-acetamido-D-hexose. Percent of 24 residues.
‰
Abundance in side chains not included in footnote z. Percent of 26
residues.
z
0, not found; o 1, less than 1%.
k
All remaining residues that occur fewer than three times.
z
The O-antigens synthesized by the ABC-transporterdependent pathway (see above) or herein tentatively assigned to that pathway are homopolymers or have only two
sugar residues in the backbone of the repeating unit (Table
4). In 1994 it was shown that in E. coli O7 (Table 5), having a
Wzy-dependent pathway, the repeating unit of the O-antigen had an N-acetylglucosamine residue at its reducing end
(Alexander & Valvano, 1994) and the authors proposed that
this pattern should also be found in other O-antigen
structures. By arranging the E. coli O-antigen structures
hitherto determined (Tables 5 and 6) with the D-GlcNAc
residue at the reducing end one readily observes that this
pattern is quite reasonable. In cases when D-GlcNAc is not
present in the polymer, D-GalNAc takes its place, in agreement with the observation that WecA can transfer either of
the N-acetylhexosamine sugars (Marolda et al., 2004). In
several of the O-antigens both amino sugars are components
of the repeating unit. In just two strains, D-FucNAc has been
found and is expected to be the sugar at the reducing end of
the repeating unit. As noted above, one of these, strain 62D1,
was recently identified as a non-E. coli species. In all but a
few cases it is possible to identify that the amino sugar at the
reducing end is 3-substituted. The other cases being the
FEMS Microbiol Rev 30 (2006) 382–403
389
Escherichia coli O-polysaccharide antigens
HO
OH
OH
O
OH
O
O
OH
HO
OH
OH
OH
OH
OH
OH
CH3
D-ribofuranose
D-fucofuranose
D-threo-pentulofuranose
OH
NHCOCH3
O
OH
O
HO
HCOHN
O
OH
OH
OH
HO
OH
OH
6-deoxy-L-talopyranose
3,6-dideoxy-L-xylo-hexose
(Colitose)
2-acetamido-2,3,6trideoxy-3-formamido-D-mannose
COOH
OH
OH
H3C
H
O
O
HO
OH
O
HO
O
AcHN
OH
OH
HO
O
OH
H
OH
COOH
H3C
4-[(R)-1-carboxyethyl]-Dglucopyranose
3-[(R)-1-carboxyethyl]-Lrhamnopyranose
4-acetamido-4,6-dideoxy-D-glucopyranose
CH3
CH2OH
COOH
COOH
HO
HO
OH
HO
O
HO
6-deoxy-D-manno-heptopyranose
H
H
OH
O
OH
HO
AcHN
H
O
OH
AcHN
OH
H
OH
OH
AcHN
N-acetyl-neuraminic acid
5,7-diacetamido-3,5,7,9tetradeoxy-L-glycero-Lmanno-nonulosonic acid
(Pseudaminic acid)
Scheme 1. Unusual glycosyl residues in Escherichia coli O-antigens.
O1A, O2 and possibly O149 antigens, where D-GlcNAc is 4substituted by a b-L-Rhap residue, or in O83 and O136,
where it is substituted by a b-D-Galp residue, i.e. the
structural element is N-acetyl-lactosamine. These results are
in good agreement with the few examples when the biological repeating unit has been determined by NMR spectroscopy, e.g. in semi-rough type of LPS containing only one
repeating unit as for E. coli O6 (Grozdanov et al., 2002). The
FEMS Microbiol Rev 30 (2006) 382–403
biological repeating unit has also been determined on
medium-sized O-antigens with a degree of polymerization
of 13 for E. coli O126 and 10 for E. coli O91 (Larsson
et al., 2004; Lycknert & Widmalm, 2004). The three-substituted D-GlcNAc residue was present in these three Opolysaccharides at the reducing end of the repeating unit.
Genetic analysis of E. coli O26 and O172 has revealed
that the second sugar is added to the D-GlcNAc-PP2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
c
390
R. Stenutz et al.
Table 3. Topology of the O-antigen repeating units
Topology
Abundance (%)
2-residue backbone
5
3
1
1
3-residue backbone
27
6
8
3
7
3
4-residue backbone
52
22
27
2
1
5-residue backbone
15
11
4
6-residue backbone
1
1
undecaprenol carrier by a UDP-L-FucNAc transferase to
form an a-(1 ! 3)-linkage (Guo et al., 2004; D’Souza
et al., 2002). Analysis of the O-antigen structures hitherto
determined indicates that in the serogroups O4, O25 and
O172 the third sugar to be added is an a-(1 ! 3)-linked
glucosyl residue, i.e. the backbone, or part of it, has the
following structure: ! X)-a-D-Glc-(1 ! 3)-a-L-FucNAc(1 ! 3)-b-D-GlcNAc(1 ! , where X represents different
linkage positions. Further genetic similarities may be present, e.g. in O4:K52, ! 2)-a-L-Rha-(1 ! 6)-a-D-Glc(1 ! 3)-a-L-FucNAc-(1 ! 3)-b-D-GlcNAc(1 ! , and O26
(e.g. without the glucosyl residue), where the last sugar is an
a-linked rhamnosyl residue, which is possibly also the case
for O25. The latter strain carries an additional D-Glc residue
that forms a substituted branch-point residue. In analogy to
the hypothesis described above, close structural relationships are observed between, for example, O6, O17, O44,
O58, O77, O78 and O88, having a Man-Man-GlcNAc
sequence at the reducing end. Although in E. coli the
numbering of serogroups is chronological, at least with
newly described strains, with the most recent ones covering
O174-O181 (Scheutz et al., 2004), subgroups are present in
some cases based on cross-reactivity, e.g. in O1, O18 and
most recently in O5, (Urbina et al., 2005) similar to the
Danish serotyping scheme for Streptococcus pneumoniae
capsular polysaccharides, which is based on cross-reactivity,
in contrast to the American system for which up to almost
100 different CPS serotypes have been described (Tomasz,
2000). In the future one may also type E. coli based on
genetic resemblance between the strains which then should
explain both structural and cross-reactivity relationships.
Furthermore, other structural similarities such as those of
blood-group determinants are present for the O86, O90,
O127 and O128 O-antigens, and these strains presumably
utilize the concept of molecular mimicry, thereby evading
the immune system of the human host (Moran et al., 1996).
In some of the E. coli strains the O-antigen structures
contain terminal glucosyl or N-acetylglycosamine residues,
e.g. in O23A, O139 and O142, as side-chains. Whether these
residues are added by a phage-induced glycosyl transferase
Table 4. O-antigens synthesised by the ABC-transporter-dependent pathway
Serogroup
O8
O9a
O9
O20ab
O20ac
O52
O97z
O101
Structure
!
!
!
!
2)-a-D-Man-(1 ! 2)-a-D-Man-(1 ! 3)-b-D-Man-(1 !
2)-a-D-Man-(1 ! 2)-a-D-Man-(1 ! 3)-a-D-Man-(1 ! 3)-a-D-Man-(1 !
2)-[a-D-Man-(1 ! 2)]2-a-D-Man-(1 ! 3)-a-D-Man-(1 ! 3)-a-D-Man-(1 !
2)-b-D-Ribf-(1 ! 4)-a-D-Gal-(1 !
a-D-Gal-(1 ! 3)
|
! 2)-b-D-Ribf-(1 ! 4)-a-D-Gal-(1 !
! 3)-b-D-Fucf-(1 ! 3)-b-D-6dmanHep2Ac-(1 !
! 3)-a-L-Rha-(1 ! 3)-b-L-Rha-(1 !
|
|
b-D-Xulf-(2 ! 2)b-D-Xulf-(2 ! 2)
! 6)-a-D-GlcNAc-(1 ! 4)-a-D-GalNAc-(1 !
Ref.
Jansson et al. (1985)
Parolis et al. (1986)
Prehm et al. (1976)
Vasil’ev & Zakharova (1976)
Vasil’ev & Zakharova (1982)
Feng et al. (2004a)
Perry & MacLean (1987)
Staaf et al. (1997)
Biosynthetic pathway proven experimentally.
w
b-D-6dmanHep2Ac is 2-O-acetyl-6-deoxy-b-D-manno-heptopyranosyl.
b-D-Xulf is b-D-threo-pentofuranosyl.
z
2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
c
FEMS Microbiol Rev 30 (2006) 382–403
FEMS Microbiol Rev 30 (2006) 382–403
c
O18A, O18ac
O16
O17
O10
O7
O6:K54
O5ab
O5ac
(strain 180/C3)
O6:K2; K13; K15
O4:K52
O4:K6
O3
O2
O1C
O1B
! 4)-a-D-GalNAc-(1 ! 3)-b-D-Man-(1 ! 4)-b-D-Man-(1 ! 3)-a-D-GlcNAc-(1 !
|
b-D-Glc-(1 ! 2)
! 4)-a-D-GalNAc-(1 ! 3)-b-D-Man-(1 ! 4)-b-D-Man-(1 ! 3)-a-D-GlcNAc-(1 !
|
b-D-GlcNAc-(1 ! 2)
a-L-Rha-(1 ! 3)
|
! 3)-b-D-Qui4NAc-(1 ! 2)-a-D-Man-(1 ! 4)-b-D-Gal-(1 ! 3)-a-D-GlcNAc-(1 !
! 3)-a-L-Rha-(1 ! 3)-a-L-Rha-(1 ! 3)-a-D-Gal-(1 ! 3)-b-D-GlcNAc-(1 !
|
a-D-Fuc4NAcyl-(1 ! 2)
Acyl = acetyl (60%) or (R)-3-hydroxybutyryl (40%)
! 2)-b-D-Galf-(1 ! 6)-a-D-Glc-(1 ! 3)-a-L-Rha2Ac-(1 ! 3)-a-D-GlcNAc-(1 !
a-D-Glc-(1 ! 6)
|
! 6)-a-D-Man-(1 ! 2)-a-D-Man-(1 ! 2)-b-D-Man-(1 ! 3)-a-D-GlcNAc(1 !
! 2)-a-L-Rha-(1 ! 6)-a-D-Glc-(1 ! 4)-a-D-Gal-(1 ! 3)-a-D-GlcNAc-(1 !
|
b-D-GlcNAc-(1 ! 3)
!
!
!
Jansson et al. (1989)
Jann et al. (1994b)
Masoud & Perry (1996)
Kenne et al. (1986)
L’vov et al. (1984)
Jann et al. (1994c)
Jansson et al. (1984)
MacLean & Perry (1997)
Urbina et al. (2005)
Jann et al. (1993)
Jann et al. (1993)
Medina et al. (1994)
Jansson et al. (1987a)
Gupta et al. (1992)
Gupta et al. (1992)
Baumann et al. (1991)
Jann et al. (1992b)
! 3)-a-L-Rha-(1 ! 3)-a-L-Rha-(1 ! 3)-b-L-Rha-(1 ! 4)-b-D-GlcNAc-(1
|
b-D-ManNAc-(1 ! 2)
! 3)-a-L-Rha-(1 ! 2)-a-L-Rha-(1 ! 2)-a-D-Gal-(1 ! 3)-b-D-GlcNAc-(1
|
b-D-ManNAc-(1 ! 2)
! 3)-a-L-Rha-(1 ! 2)-a-L-Rha-(1 ! 3)-a-D-Gal-(1 ! 3)-b-D-GlcNAc-(1
|
b-D-ManNAc-(1 ! 2)
! 3)-a-L-Rha-(1 ! 2)-a-L-Rha-(1 ! 3)-b-L-Rha-(1 ! 4)-b-D-GlcNAc-(1
|
a-D-Fuc3NAc-(1 ! 2)
b-L-RhaNAc(1 ! 4)
a-D-Glc-(1 ! 4)
|
|
! 3)-b-D-GlcNAc-(1 ! 3)-a-D-Gal-(1 ! 3)-b-D-GlcNAc-(1 !
! 2)-a-L-Rha-(1 ! 6)-a-D-Glc-(1 ! 3)-a-L-FucNAc-(1 ! 3)-b-D-GlcNAc(1 !
a-D-Glc-(1 ! 3)
|
! 2)-a-L-Rha-(1 ! 6)-a-D-Glc-(1 ! 3)-a-L-FucNAc-(1 ! 3)-b-D-GlcNAc(1 !
! 4)-b-D-Qui3NAc-(1 ! 3)-b-D-Ribf-(1 ! 4)-b-D-Gal-(1 ! 3)-a-D-GalNAc(1 !
! 2)-b-D-Qui3NAc-(1 ! 3)-b-D-Ribf-(1 ! 4)-b-D-Gal-(1 ! 3)-a-D-GalNAc(1 !
O1A, O1A1
!
Ref.
Structure
Serogroup
Table 5. O-antigens synthesised by the polymerase-dependent pathway with four or less residues in the backbone
Escherichia coli O-polysaccharide antigens
391
2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
c
2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
O56
O45
O45rel
O55
O26
O28
O44
O25
O24
O23A
O21
O18B1
Gamian et al. (1994)
Jann et al. (1995)
Jann et al. (1995)
Lindberg et al. (1981)
Manca et al. (1996)
Rundlöf et al. (1996)
Staaf et al. (1995)
Kenne et al. (1983a)
Torgov et al. (1995)
Bartelt et al. (1993)
Staaf et al. (1999a)
Jann et al. (1992a)
Jann et al. (1992a)
Jann et al. (1992a)
a-D-Glc-(1 ! 6)
|
! 2)-a-L-Rha-(1 ! 6)-a-D-Glc-(1 ! 4)-a-D-Gal-(1 ! 3)-a-D-GlcNAc-(1 !
|
b-D-GlcNAc-(1 ! 3)
! 3)-a-L-Rha-(1 ! 6)-a-D-Glc-(1 ! 4)-a-D-Gal-(1 ! 3)-a-D-GlcNAc-(1 !
|
b-D-Glc-(1 ! 3)
a-D-Glc-(1 ! 4)
|
! 3)-a-L-Rha-(1 ! 6)-a-D-Glc-(1 ! 4)-a-D-Gal-(1 ! 3)-a-D-GlcNAc-(1 !
|
b-D-Glc-(1 ! 3)
b-D-Gal-(1 ! 4)
|
! 3)-b-D-Gal-(1 ! 4)-b-D-Glc-(1 ! 3)-b-D-GalNAc-(1 !
|
b-D-GlcNAc-(1 ! 2)
a-D-Glc-(1 ! 6)
|
! 6)-a-D-Glc-(1 ! 4)-b-D-Gal-(1 ! 3)-a-D-GalNAc-(1 ! 3)-b-D-GlcNAc-(1 !
|
b-D-GlcNAc(1 ! 3)
! 7)-a-Neu5Ac-(2 ! 3)-b-D-Glc-(1 ! 3)-b-D-GalNAc-(1 !
|
a-D-Glc-(1 ! 2)
b-D-Glc-(1 ! 6)
|
! 4)-a-D-Glc-(1 ! 3)-a-L-FucNAc-(1 ! 3)-b-D-GlcNAc-(1 !
|
a-L-Rha-(1 ! 3)
! 3)-a-L-Rha-(1 ! 4)-a-L-FucNAc-(1 ! 3)-b-D-GlcNAc-(1 !
! 2)-(R)-Gro-1-P ! 4)-b-D-GlcNAc-(1 ! 3)-b-D-Galf2Ac-(1 ! 3)-a-D-GlcNAc-(1 !
a-D-Glc-(1 ! 4)
|
! 6)-a-D-Man-(1 ! 2)-a-D-Man-(1 ! 2)-b-D-Man-(1 ! 3)-a-D-GlcNAc(1 !
! 2)-b-D-Glc-(1 ! 3)-a-L-6dTal2Ac-(1 ! 3)-a-D-FucNAc-(1 !
! 2)-b-D-Glc-(1 ! 3)-a-L-6dTal2Ac-(1 ! 3)-b-D-GlcNAc-(1 !
! 6)-b-D-GlcNAc-(1 ! 3)-a-D-Gal-(1 ! 3)-b-D-GalNAc-(1 !
|
a-Col-(1 ! 2)-b-D-Gal-(1 ! 3)
! 7)-a-Neu5Ac-(2 ! 3)-b-D-Glc-(1 ! 3)-b-D-GlcNAc-(1 !
|
a-D-Gal-(1 ! 2)
O18A1
O18B
Ref.
Structure
Serogroup
Table 5. Continued.
392
R. Stenutz et al.
FEMS Microbiol Rev 30 (2006) 382–403
FEMS Microbiol Rev 30 (2006) 382–403
c
O125
O121
O124
O114
O119
O113
O90
O98
O104
O111
O88
O77
O78
O86
O75
O69
O73
(Strain 73-1)
Kjellberg et al. (1996)
Parolis et al. (1997)
Dmitriev et al. (1976)
Dmitriev et al. (1983)
Anderson et al. (1992)
Parolis & Parolis (1995)
Ratnayake et al. (1994b)
Marsden et al. (1994)
Gamian et al. (1992)
Eklund et al. (1978)
Torgov et al. (1996)
Yildirim et al. (2001)
Jansson et al. (1987b)
Andersson et al. (1989)
Erbing et al. (1978)
Erbing et al. (1977)
Weintraub et al. (1993)
Perry et al. (1993)
3-O-[(R)-1-carboxyethyl]-a-L-Rha -(1 ! 3)
|
! 4)-a-D-Man-(1 ! 4)-a-D-Man2Ac-(1 ! 3)-b-D-GlcNAc-(1 !
b-D-Gal-(1 ! 6)
|
! 3)-a-D-ManNAc-(1 ! 3)-b-D-GlcA-(1 ! 3)-b-D-Gal-(1 ! 3)-b-D-GlcNAc(1 !
! 2)-a-L-Rha-(1 ! 2)-a-L-Rha-(1 ! 2)-a-D-Gal-(1 ! 3)-b-D-GlcNAc-(1 !
a-D-Glc-(1 ! 3)
|
! 4)-a-D-Man-(1 ! 2)-a-D-Man-(1 ! 2)-b-D-Man-(1 ! 3)-a-D-GalNAc(1 !
b-D-Man-(1 ! 4)
|
! 3)-a-D-Gal-(1 ! 4)-a-L-Rha-(1 ! 3)-b-D-GlcNAc-(1 !
! 6)-a-D-Man-(1 ! 2)-a-D-Man-(1 ! 2)-b-D-Man-(1 ! 3)-a-D-GlcNAc(1 !
! 4)-b-D-GlcNAc-(1 ! 4)-b-D-Man-(1 ! 4)-a-D-Man-(1 ! 3)-b-D-GlcNAc-(1 !
a-D-Gal-(1 ! 3)
|
! 4)-a-L-Fuc-(1 ! 2)-b-D-Gal-(1 ! 3)-a-D-GalNAc-(1 ! 3)-b-D-GalNAc-(1 !
a-L-6dTal-(1 ! 3)
|
! 4)-a-D-Man-(1 ! 3)-a-D-Man-(1 ! 3)-b-D-GlcNAc-(1 !
! 4)-a-L-Fuc2/3Ac-(1 ! 2)-b-D-Gal-(1 ! 3)-a-D-GalNAc-(1 ! 3)-b-D-GalNAc-(1 !
! 3)-a-L-QuiNAc-(1 ! 4)-a-D-GalNAcA-(1 ! 3)-a-L-QuiNAc-(1 ! 3)-b-D-GlcNAc-(1 !
! 4)-a-D-Gal-(1 ! 4)-a-Neu5,7,9Ac3-(2 ! 3)-b-D-Gal-(1 ! 3)-b-D-GalNAc-(1 !
a-Col-(1 ! 6)
|
! 4)-a-D-Glc-(1 ! 4)-a-D-Gal-(1 ! 3)-b-D-GlcNAc-(1 !
|
a-Col-(1 ! 3)
! 4)-a-D-GalNAc-(1 ! 4)-a-D-GalA-(1 ! 3)-a-D-Gal-(1 ! 3)-b-D-GlcNAc-(1 !
|
b-D-Gal-(1 ! 3)
! 4)-b-D-Qui3N(N-acetyl-L-seryl)-(1 ! 3)-b-D-Ribf-(1 ! 4)-b-D-Gal-(1 ! 3)-a-D-GlcNAc(1 !
b-D-RhaNAc3NFo-(1 ! 3)
|
! 2)-b-D-Man-(1 ! 3)-a-D-Gal-(1 ! 4)-a-L-Rha-(1 ! 3)-a-D-GlcNAc-(1 !
! 3)-b-D-Qui4N(N-acetyl-glycyl)-(1 ! 4)-a-D-GalNAc3AcA6N-(1 ! 4)-a-D-GalNAcA-(1 ! 3)-a-D-GlcNAc-(1 !
4-O-[(R)-1-carboxyethyl]-b-D-Glc-(1 ! 6)-a-D-Glc(1 ! 4)
|
! 3)-a-D-Gal-(1 ! 6)-b-D-Galf-(1 ! 3)-b-D-GalNAc-(1 !
a-D-Glc-(1 ! 3)
|
! 4)-b-D-GalNAc-(1 ! 2)-a-D-Man-(1 ! 3)-a-L-Fuc-(1 ! 3)-a-D-GalNAc-(1 !
|
b-D-Gal-(1 ! 3)
O58
O64
Ref.
Dmitriev et al. (1977)
Structure
Serogroup
Escherichia coli O-polysaccharide antigens
393
2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
c
2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
62D1
O173
O164
O159
O158
O157
O152
O147
O149
O143
O142
O138
O141
O136
a-D-Glc-(1 ! 6)
|
! 4)-a-D-Glc-(1 ! 3)-a-D-GalNAc-(1 ! 3)-b-D-GalNAc-(1 !
|
a-L-Rha-(1 ! 3)
a-L-Fuc-(1 ! 4)
|
! 3)-b-D-GlcNAc-(1 ! 4)-a-D-GalA-(1 ! 3)-a-L-Fuc-(1 ! 3)-b-D-GlcNAc-(1 !
b-D-Glc-(1 ! 6)-a-D-Glc(1 ! 4)
|
! 3)-b-D-Gal-(1 ! 6)-b-D-Galf-(1 ! 3)-b-D-GalNAc-(1 !
a-L-Fuc-(1 ! 4)
|
! 3)-a-D-Glc-(1-P ! 6)-a-D-Glc-(1 ! 2)-b-D-Glc-(1 ! 3)-b-D-GlcNAc-(1 !
a-D-Gal(1 ! 6)
|
! 2)-b-D-Qui3NAc-(1 ! 3)-a-L-Rha-(1 ! 3)-b-D-Gal-(1 ! 3)-a-D-FucNAc-(1 !
Suggested as Erwinia herbicola
! 2)-b-D-Man-(1 ! 3)-b-D-Gal-(1 ! 3)-a-D-GlcNAc-(1 ! 3)-b-D-GlcNAc-(1 !
|
a-L-Fuc-(1 ! 2)
! 2)-a-L-Fuc-(1 ! 2)-b-D-Gal-(1 ! 3)-a-D-GalNAc-(1 ! 3)-a-D-GalNAc-(1 !
a-L-Fuc-(1 ! 2)
|
! 6)-b-D-Gal-(1 ! 3)-b-D-GalNAc-(1 ! 4)-a-D-Gal-(1 ! 3)-b-D-GalNAc-(1 !
! 4)-b-Pse5Ac7Ac-(2 ! 4)-b-D-Gal-(1 ! 4)-b-D-GlcNAc-(1 !
b-Pse5Ac7Ac = 5,7-diacetamido-3,5,7,9-tetradeoxy-L-glycero-b-L-manno-nonulosonic acid
! 2)-a-L-Rha-(1 ! 3)-a-L-Rha-(1 ! 4)-a-D-GalNAcA-(1 ! 3)-b-D-GlcNAc-(1 !
a-L-Rha-(1 ! 3)
|
! 4)-a-D-Man-(1 ! 3)-a-D-Man6Ac-(1 ! 3)-b-D-GlcNAc-(1 !
|
b-D-GlcA-(1 ! 2)
! 2)-a-L-Rha-(1 ! 6)-a-D-GalNAc-(1 ! 4)-a-D-GalNAc-(1 ! 3)-a-D-GalNAc-(1 !
|
b-D-GlcNAc-(1 ! 3)
! 2)-b-D-GalA6R3,4Ac-(1 ! 3)-a-D-GalNAc-(1 ! 4)-b-D-GlcA-(1 ! 3)-b-D-GlcNAc-(1 !
R = 1,3-dihydroxy-2-propylamino
! 2)-a-L-Rha-(1 ! 2)-a-L-Rha-(1 ! 4)-b-D-GalA-(1 ! 3)-b-D-GalNAc-(1 !
! 3)-b-D-GlcNAc-(S)-4,6Py-(1 ! 3)-b-L-Rha-(1 ! 4)-b-D-GlcNAc-(1 !
(S)-4,6Py = 4,6-O-[(S)-1-carboxyethylidene]b-L-Rha-(1 ! 4)
|
! 3)-a-D-GlcNAc-(1-P ! 6)-a-D-Glc-(1 ! 2)-b-D-Glc-(1 ! 3)-b-D-GlcNAc-(1 !
! 2)-a-D-Rha4NAc-(1 ! 3)-a-L-Fuc-(1 ! 4)-b-D-Glc-(1 ! 3)-a-D-GalNAc-(1 !
O126
O127
O128
Structure
Serogroup
Table 5. Continued.
Staaf et al. (1999b)
Linnerborg et al. (1999b)
Linnerborg et al. (1999a)
Linnerborg et al. (1999c)
Datta et al. (1999)
Nishiuchi et al. (2002)
Nishiuchi et al. (2000)
Perry et al. (1986)
Olsson et al. (2005)
Hygge Blackeman et al. (1998)
Adeyeye et al. (1988)
Landersjö et al. (1996)
Landersjö et al. (1997)
Linnerborg et al. (1997a)
Färnbäck et al. (1998)
Staaf et al. (1999c)
Widmalm & Leontein, (1993)
Sengupta et al. (1995)
Larsson et al. (2004)
Ref.
394
R. Stenutz et al.
FEMS Microbiol Rev 30 (2006) 382–403
FEMS Microbiol Rev 30 (2006) 382–403
O172
O153
O167
O116
O117
O139
O105
Landersjö et al. (2001)
Ratnayake et al. (1994a)
Linnerborg et al. (1997b)
Leslie et al. (1999)
Leslie et al. (2000)
Marie et al. (1998)
Tao et al. (2004)
Perry & MacLean (1999)
Jann et al. (1995)
Jann et al. (1994a)
Kjellberg et al. (1999)
! 6)-a-D-Glc-(1 ! 4)-b-D-GlcA-(1 ! 4)-b-D-GalNAc3Ac-(1 ! 3)-a-D-Gal-(1 ! 3)-b-D-GalNAc-(1 !
! 3)-a-L-Rha-(1 ! 2)-a-L-Rha-(1 ! 3)-a-L-Rha-(1 ! 2)-a-L-Rha-(1 ! 3)-b-D-GlcNAc-(1 !
|
a-D-GalNAcA6N-(1 ! 2)
! 2)-b-D-Qui3NAc-(1 ! 4)-a-D-GalA6N-(1 ! 4)-a-D-GalNAc-(1 ! 4)-b-D-GalA-(1 ! 3)-a-D-GlcNAc-(1 !
! 2)-b-D-Man-(1 ! 3)-a-D-GlcNAc-(1 ! 2)-b-D-Glc3Ac-(1 ! 3)-a-L-6dTal-(1 ! 3)-a-D-GlcNAc(1 !
! 6)-a-D-Glc-(1 ! 4)-b-D-GlcA-(1 ! 6)-b-D-Gal-(1 ! 4)-b-D-Gal-(1 ! 4)-b-D-GlcNAc-(1 !
! 4)-a-D-Qui3NAcyl-(1 ! 4)-b-D-Gal-(1 ! 4)-b-D-GlcNAc-(1 ! 4)-b-D-GlcA6NGly-(1 ! 3)-b-D-GlcNAc-(1 !
Acyl = (R)-3-hydroxybutyryl
b-D-Ribf-(1 ! 3)
|
! 4)-a-D-GlcA2Ac3Ac-(1 ! 2)-a-L-Rha4Ac-(1 ! 3)-b-L-Rha-(1 ! 4)-b-L-Rha-(1 ! 3)-b-D-GlcNAc6Ac-(1 !
! 2)-b-D-Qui4NAc-(1 ! 6)-a-D-GlcNAc-(1 ! 4)-a-D-GalNAc-(1 ! 4)-a-D-GalA-(1 ! 3)-b-D-GlcNAc-(1 !
! 4)-b-D-GalNAc-(1 ! 3)-a-L-Rha-(1 ! 4)-a-D-Glc-(1 ! 4)-b-D-Gal-(1 ! 3)-a-D-GalNAc-(1 !
b-D-Glc-(1 ! 3)
|
! 3)-a-L-Rha-(1 ! 4)-a-D-GalA-(1 ! 2)-a-L-Rha-(1 ! 3)-a-L-Rha-(1 ! 2)-a-L-Rha-(1 ! 3)-a-D-GlcNAc-(1 !
! 2)-b-D-Ribf-(1 ! 4)-b-D-Gal-(1 ! 4)-a-D-GlcNAc-(1 ! 4)-b-D-Gal-(1 ! 3)-a-D-GlcNAc-(1 !
a-D-Galf-(1 ! 4)
|
! 2)-b-D-GalA6N(L)Ala-(1 ! 3)-a-D-GlcNAc-(1 ! 2)-b-D-Galf-(1 ! 5)-b-D-Galf-(1 ! 3)-b-D-GlcNAc-(1 !
! 3)-a-L-FucNAc-(1 ! 4)-a-D-Glc6Ac-(1-P ! 4)-a-D-Glc-(1 ! 3)-a-L-FucNAc-(1 ! 3)-a-D-GlcNAc-(1 !
O22
O35
O65
O66
O83
O91
Ref.
Bartelt et al. (1994)
Rundlöf et al. (1998)
Structure
Serogroup
Table 6. O-antigens synthesized by the polymerase-dependent pathway with five or six residues in the backbone
Escherichia coli O-polysaccharide antigens
395
c
2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
396
machinery or by another mechanism is of great interest for
future genetic studies as the positioning of the side-chain
onto the structure differs and sometimes leads to a doubly
branched residue, e.g. in O141. In many cases the repeating
unit is formed and structurally determined by the polymerization process, e.g. in O55 and O164, often occurring at the
penultimate sugar residue of the linear undecaprenol-PPoligosaccharide leading to a single sugar residue in the sidechain, e.g. in O35, O113, O152, O159 and O167.
The O-antigen of Shigella boydii type 13 was recently both
structurally and genetically characterized (Feng et al.,
2004b). Although this strain is more distantly related to E.
coli and other Shigella species, its O-antigen shows a quite
close structural resemblance to that of E. coli O172. The
linear pentasaccharide of S. boydii type 13 has the following
chemical structure: ! 3)-a-L-QuipNAc-(1 ! 4)-a-D-Glcp(1 ! P-4)-a-D-GlcpNAc-(1 ! 3)-a-L-QuipNAc-(1 ! 3)-aD-GlcpNAc-(1 ! , in which the 4-linked GlcNAc residue is
6-O-acetylated to 15%. Based on biosynthetic considerations where an N-acetylglycosamine residue should be present
at the reducing end, two possibilities were suggested for the
biological repeating unit, i.e. the one above or the frameshifted one with the 4-linked GlcNAc residue at the reducing
end. From the structural results presented in this article we
propose that the biological repeating unit of the O-antigen
from S. boydii type 13 has a 3-linked GlcNAc residue at the
reducing end as presented in the above structure. In both the
E. coli O172 and the S. boydii type 13 O-antigens the glucosyl1-phosphoryl residue is the penultimate one (as presented) in
the assembled linear undecaprenol-PP-oligosaccharide. The
O-antigens of E. coli O152 and O173 have branched structures with one sugar residue as the side-chain and, most
notably, the branching sugar is a glycosyl-1-phosphoryl
residue being the penultimate one, suggesting similar biosynthetic pathways. Future detailed investigations will clarify the
whole assembly of these E. coli O-antigen units and that of S.
boydii type 13.
Concluding remarks
Members of the species Escherichia coli range from completely harmless to life-threatening microorganisms. The differences are based on particular virulence factors that certain
strains may have acquired. These factors may be toxins or
surface structures that enable the bacterium to adhere to
mammalian cells or to evade the immune system. The
typing of E. coli is often based on detection of different
surface molecules using specific antibodies. The O-antigen
present in the lipopolysaccharide is one of the molecules
used in serotyping. As of today, more than 180 different Oserotypes have been described but not even half of them
have been structurally elucidated.
2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
c
R. Stenutz et al.
The E. coli database implemented facilitates a more rapid
identification of strains that are difficult to type or suggests
similarities to previously determined O-antigens in the case
of novel isolates. Analysis of the O-antigen structures
revealed that 3-linked N-acetylglucosamine or N-acetylgalactosamine residues should be present at the reducing end
of the biological repeating unit, in accordance with NMR
spectroscopy and genetic data. In a limited number of cases,
a 4-linked N-acetylglucosamine residue is instead observed.
The topology with four sugar residues in the backbone of
the O-antigen is present in half of the hitherto determined
structures. Future structural studies should be combined
with genetic analysis of the O-antigen cluster to facilitate
insight into structural patterns and biosynthetic pathways.
As part of this effort, amino acid sequences of flippases and
polymerases are being added as elements of the entries in the
database.
Note
This review covers structures reported up to early 2005. In
addition, recently reported E. coli O-antigen structures are
those of serogroups O178 (Ali et al., 2005) and O145 (Feng
et al., 2005). Noteworthy is also the fact that phenotypically
rough E. coli K12 have been genetically complemented to
produce its O-antigen, an O16 variant with cross-reactivity
to O17 (Liu & Reeves, 1994; Stevenson et al., 1994).
Acknowledgements
This work was supported by grants from the Swedish
Research Council and from the Swedish Agency for Research
Cooperation with Developing Countries SIDA/SAREC.
References
Adeyeye A, Jansson PE, Lindberg B, Abaas S & Svenson SB (1988)
Structural studies of the Escherichia coli O-149 O-antigen
polysaccharide. Carbohydr Res 176: 231–236.
Albert MJ, Faruque SM, Ansaruzzaman M, Islam MM, Haider K,
Alam K, Kabir I & Robins-Browne R (1992) Sharing of
virulence-associated properties at the phenotypic and genetic
levels between enteropathogenic Escherichia coli and Hafnia
alvei. J Med Microbiol 37: 310–314.
Alexander DC & Valvano MA (1994) Role of the rfe gene in the
biosynthesis of the Escherichia coli O7-specific
lipopolysaccharide and other O-specific polysaccharides
containing N-acetylglucosamine. J Bacteriol 176: 7079–7084.
Ali T, Urbina F, Weintraub A & Widmalm G (2005) Structural
studies of the O-antigenic polysaccharides from the
enteroaggregative Escherichia coli strain 522/C1 and the
international type strain from Escherichia coli O178. Carbohydr
Res 340: 2010–2014.
FEMS Microbiol Rev 30 (2006) 382–403
397
Escherichia coli O-polysaccharide antigens
Andersson M, Carlin N, Leontein K, Lindquist U & Slettengren K
(1989) Structural studies of the O-antigenic polysaccharide of
Escherichia coli O86, which possesses blood-group B activity.
Carbohydr Res 185: 211–223.
Anderson AN, Richards JC & Perry MB (1992) Structure of the
O-antigen of Escherichia coli O119 lipopolysaccharide.
Carbohydr Res 237: 249–262.
Arduino RC & DuPont HL (1993) Travellers’ diarrhoea. Baillieres
Clin Gastroenterol 7: 365–385.
Aucken HM & Pitt TL (1991) Serological relationships of the O
antigens of Klebsiella pneumoniae O5, Escherichia coli O8 and a
new O serotype of Serratia marcescens. FEMS Microbiol Lett 64:
93–97.
Bartelt M, Shashkov AS, Kochanowski H, Jann B & Jann K (1993)
Structure of the O-specific polysaccharide of the O23 antigen
(LPS) from Escherichia coli O23:K?H16. Carbohydr Res 248:
233–240.
Bartelt M, Shashkov AS, Kochanowski H, Jann B & Jann K (1994)
Structure of the O-specific polysaccharide of the O22-antigen
(LPS) from Escherichia coli O22:K13. Carbohydr Res 254:
203–212.
Baudry B, Maurelli AT, Clerc P, Sadoff JC & Sansonetti PJ (1987)
Localization of plasmid loci necessary for the entry of Shigella
flexneri into HeLa cells, and characterization of one locus
encoding four immunogenic polypeptides. J Gen Microbiol
133: 3403–3413.
Baumann H, Jansson PE, Kenne L & Widmalm G (1991)
Structural studies of the Escherichia coli O1A Opolysaccharide, using the computer program CASPER.
Carbohydr Res 211: 183–190.
Bettelheim KA, Evangelidis H, Pearce JL, Sowers E & Strockbine
NA (1993) Isolation of a Citrobacter freundii strain which
carries the Escherichia coli O157 antigen. J Clin Microbiol 31:
760–761.
Bhan MK, Bhandari N, Sazawal S, Clemens J, Raj P, Levine MM &
Kaper JB (1989a) Descriptive epidemiology of persistent
diarrhoea among young children in rural northern India. Bull
World Health Organ 67: 281–288.
Bhan MK, Raj P, Levine MM, Kaper JB, Bhandari N, Srivastava R,
Kumar R & Sazawal S (1989b) Enteroaggregative Escherichia
coli associated with persistent diarrhea in a cohort of rural
children in India. J Infect Dis 159: 1061–1064.
Bhan MK, Sazawal S, Raj P, Bhandari N, Kumar R, Bhardwaj Y,
Shrivastava R & Bhatnagar S (1989c) Aggregative
Escherichia coli, Salmonella, and Shigella are associated
with increasing duration of diarrhea. Indian J Pediatr 56:
81–86.
Bhatnagar S, Bhan MK, Sommerfelt H, Sazawal S, Kumar R &
Saini S (1993) Enteroaggregative Escherichia coli may be a new
pathogen causing acute and persistent diarrhea. Scand J Infect
Dis 25: 579–583.
Bilge SS, Clausen CR, Lau W & Moseley SL (1989) Molecular
characterization of a fimbrial adhesin, F1845, mediating
diffuse adherence of diarrhea-associated Escherichia coli to
hep-2 cells. J Bacteriol 171: 4281–4289.
FEMS Microbiol Rev 30 (2006) 382–403
Bilge SS, Apostol JM Jr, Aldape MA & Moseley SL (1993a) mRNA
processing independent of RNase III and RNase E in the
expression of the F1845 fimbrial adhesin of Escherichia coli.
Proc Natl Acad Sci USA 90: 1455–1459.
Bilge SS, Apostol JM Jr, Fullner KJ & Moseley SL (1993b)
Transcriptional organization of the F1845 fimbrial adhesin
determinant of Escherichia coli. Mol Microbiol 7: 993–1006.
Black RE (1990) Epidemiology of travelers’ diarrhea and relative
importance of various pathogens. Rev Infect Dis 12(Suppl. 1):
S73–S79.
Black RE (1993) Epidemiology of diarrhoeal disease: implications
for control by vaccines. Vaccine 11: 100–106.
Bouzari S, Jafari A, Farhoudi-Moghaddam AA, Shokouhi F &
Parsi M (1994) Adherence of non-enteropathogenic
Escherichia coli to HeLa cells. J Med Microbiol 40: 95–97.
Boyce TG, Swerdlow DL & Griffin PM (1995) Escherichia coli
O157:H7 and the hemolytic-uremic syndrome. N Engl J Med
333: 364–368.
Brade H, Opal SM, Vogel SN & Morrison DC (eds) (1999)
Endotoxin in Health and Disease. Marcel Dekker, Inc.,
New York.
Bundle DR, Gerken M & Perry MB (1986) Two-dimensional
nuclear magnetic resonance at 500 MHz: the structural
elucidation of a Salmonella serogroup N polysaccharide
antigen. Can J Chem 64: 255–264.
Campos LC, Franzolin MR & Trabulsi LR (2004) Diarrheagenic
Escherichia coli categories among the traditional
enteropathogenic E. coli O serogroups. Mem Inst Oswaldo Cruz
99: 545–552.
Chen HD & Frankel G (2005) Enteropathogenic Escherichia coli:
unravelling pathogenesis. FEMS Microbiol Rev 29: 83–98.
Cravioto A, Reyes RE, Ortega R, Fernandez G, Hernandez R &
Lopez D (1988) Prospective study of diarrhoeal disease in a
cohort of rural Mexican children: incidence and isolated
pathogens during the first two years of life. Epidemiol Infect
101: 123–134.
Cravioto A, Reyes RE, Trujillo F, Uribe F, Navarro A, De La Roca
JM, Hernandez JM, Perez G & Vazquez V (1990) Risk of
diarrhea during the first year of life associated with initial and
subsequent colonization by specific enteropathogens. Am J
Epidemiol 131: 886–904.
Cravioto A, Tello A, Navarro A, Ruiz J, Villafan H, Uribe F &
Eslava C (1991) Association of Escherichia coli HEp-2
adherence patterns with type and duration of diarrhoea.
Lancet 337: 262–264.
Darfeuille-Michaud A (2002) Adherent-invasive Escherichia coli:
a putative new E. coli pathotype associated with Crohn’s
disease. Int J Med Microbiol 292: 185–193.
Datta AK, Basu S & Roy N (1999) Chemical and
immunochemical studies of the O-antigen from
enteropathogenic Escherichia coli O158 lipopolysaccharide.
Carbohydr Res 322: 219–227.
Dawson KG, Emerson JC & Burns JL (1999) Fifteen years of
experience with bacterial meningitis. Pediatr Infect Dis J 18:
816–822.
2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
c
398
De Rycke J, Milon A & Oswald E (1999) Necrotoxic Escherichia
coli (NTEC): two emerging categories of human and animal
pathogens. Vet Res 30: 221–233.
Dmitriev BA, Lvov VL, Kochetkov NK, Jann B & Jann K (1976)
Cell-wall lipopolysaccharide of the ‘Shigella-like’ Escherichia
coli O124. Structure of the polysaccharide chain. Eur J Biochem
64: 491–498.
Dmitriev BA, Knirel YA, Kochetkov NK, Jann B & Jann K (1977)
Cell-wall lipopolysaccharide of the ‘Shigella-like’ Escherichia
coli O58. Structure of the polysaccharide chain. Eur J Biochem
79: 111–115.
Dmitriev BA, Lvov V, Tochtamysheva NV, Shashkov AS,
Kochetkov NK, Jann B & Jann K (1983) Cell-wall
lipopolysaccharide of Escherichia coli O114:H2. Structure of
the polysaccharide chain. Eur J Biochem 134: 517–521.
Donnenberg MS & Nataro JP (1995) Methods for studying
adhesion of diarrheagenic Escherichia coli. Methods Enzymol
253: 324–336.
D’Souza JM, Wang L & Reeves P (2002) Sequence of the
Escherichia coli O26 O antigen gene cluster and identification
of O26 specific genes. Gene 297: 123–127.
DuPont HL & Ericsson CD (1993) Prevention and treatment of
traveler’s diarrhea. N Engl J Med 328: 1821–1827.
Dutta S, Chatterjee A, Dutta P, Rajendran K, Roy S, Pramanik KC
& Bhattacharya SK (2001) Sensitivity and performance
characteristics of a direct PCR with stool samples in
comparison to conventional techniques for diagnosis of
Shigella and enteroinvasive Escherichia coli infection in
children with acute diarrhoea in Calcutta, India. J Med
Microbiol 50: 667–674.
Eklund K, Garegg PJ, Kenne L, Lindberg AA & Lindberg B (1978)
Structural studies on the Escherichia coli O111
lipopolysaccharide. Abstracts of the IXth International
Symposium of Carbohydrate Chemistry, London.
Erbing C, Kenne L & Lindberg B (1977) Structural studies of the
O-specific side-chains of the cell-wall lipopolysaccharide from
Escherichia coli O69. Carbohydr Res 56: 371–376.
Erbing C, Kenne L, Lindberg B & Hammarström S (1978)
Structure of the O-specific side-chain of the Escherichia coli
O75 lipopolysaccharide: a revision. Carbohydr Res 60:
400–403.
Ewing WH (1986) The genus Escherichia. Edwards and Ewing’s
identification of Enterobacteriaceae, 4th edn (Edwards PR &
Ewing WH, eds) pp. 93–134. Elsevier Science Publishing Co.,
Inc., New York.
Ezawa A, Gocho F, Saitoh M, Tamura T, Kawata K, Takahashi T &
Kikuchi N (2004) A three-year study of enterohemorrhagic
Escherichia coli O157 on a farm in Japan. J Vet Med Sci 66:
779–784.
Fang GD, Lima AA, Martins CV, Nataro JP & Guerrant RL (1995)
Etiology and epidemiology of persistent diarrhea in
northeastern Brazil: a hospital-based, prospective, case-control
study. J Pediatr Gastroenterol Nutr 21: 137–144.
2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
c
R. Stenutz et al.
Färnbäck M, Weintraub A & Widmalm G (1998) Structural
determination of the O-antigenic polysaccharide from
Escherichia coli O141. Eur J Biochem 254: 168–171.
Feng L, Senchenkova SN, Yang J, et al. (2004a) Synthesis of the
heteropolysaccharide O antigen of Escherichia coli O52
requires an ABC transporter: structural and genetic evidence.
J Bacteriol 186: 4510–4519.
Feng L, Senchenkova SN, Yang J, Shashkov AS, Tao J, Guo H,
Zhao G, Knirel YA, Reeves P & Wang L (2004b) Structural and
genetic characterization of the Shigella boydii type 13 O
antigen. J Bacteriol 186: 383–392.
Feng L, Senchenkova SN, Tao J, Shashkov AS, Liu B, Shevelev SD,
Reeves PR, Xu J, Knirel YA & Wang L (2005) Structural and
genetic characterization of enterohemorrhagic Escherichia coli
O145 O antigen and development of an O145 serogroupspecific PCR assay. J Bacteriol 187: 758–764.
Flores Abuxapqui JJ, Suarez Hoil GJ, Heredia Navarrete MR, Puc
Franco MA & Vivas Rosel ML (1999) Four biochemical tests
for identification of probable enteroinvasive Escherichia coli
strains. Rev Latinoam Microbiol 41: 259–261.
Gamian A, Romanowska E, Ulrich J & Defaye J (1992) The
structure of the sialic acid-containing Escherichia coli O104 Ospecific polysaccharide and its linkage to the core region in
lipopolysaccharide. Carbohydr Res 236: 195–208.
Gamian A, Kenne L, Mieszala M, Ulrich J & Defaye J (1994)
Structure of the Escherichia coli O24 and O56 O-specific sialicacid-containing polysaccharides and linkage of these
structures to the core region in lipopolysaccharides. Eur J
Biochem 225: 1211–1220.
Germani Y, Begaud E, Duval P & Le Bouguenec C (1996)
Prevalence of enteropathogenic, enteroaggregative, and
diffusely adherent Escherichia coli among isolates from
children with diarrhea in new Caledonia. J Infect Dis 174:
1124–1126.
Gioffre A, Meichtri L, Zumarraga M, Rodriguez R & Cataldi A
(2004) Evaluation of a QIAamp DNA stool purification kit for
Shiga-toxigenic Escherichia coli detection in bovine fecal swabs
by PCR. Rev Argent Microbiol 36: 1–5.
Giron JA, Jones T, Millan-Velasco F, et al. (1991) Diffuse-adhering
Escherichia coli (DAEC) as a putative cause of diarrhea in
Mayan children in Mexico. J Infect Dis 163: 507–513.
Goldberg MB & Sansonetti PJ (1993) Shigella subversion of the
cellular cytoskeleton: a strategy for epithelial colonization.
Infect Immun 61: 4941–4946.
Gomes TA, Blake PA & Trabulsi LR (1989) Prevalence of
Escherichia coli strains with localized, diffuse, and aggregative
adherence to HeLa cells in infants with diarrhea and matched
controls. J Clin Microbiol 27: 266–269.
Gomes TA, Rassi V, Mac Donald KL, et al. (1991)
Enteropathogens associated with acute diarrheal disease in
urban infants in Sao Paulo, Brazil. J Infect Dis 164: 331–337.
Gonzalez R, Diaz C, Marino M, Cloralt R, Pequeneze M & PerezSchael I (1997) Age-specific prevalence of Escherichia coli with
localized and aggregative adherence in Venezuelan infants with
acute diarrhea. J Clin Microbiol 35: 1103–1107.
FEMS Microbiol Rev 30 (2006) 382–403
399
Escherichia coli O-polysaccharide antigens
Grimm LM, Goldoft M, Kobayashi J, Lewis JH, Alfi D, Perdichizzi
AM, Tarr PI, Ongerth JE, Moseley SL & Samadpour M (1995)
Molecular epidemiology of a fast-food restaurant-associated
outbreak of Escherichia coli O157:H7 in Washington State. J
Clin Microbiol 33: 2155–2158.
Grozdanov L, Zähringer U, Blum-Oehler G, et al. (2002) A single
nucleotide exchange in the wzy gene is responsible for the
semirough O6 lipopolysaccharide phenotype and serum
sensitivity of Escherichia coli strain Nissle 1917. J Bacteriol 184:
5912–5925.
Gunzburg ST, Chang BJ, Elliott SJ, Burke V & Gracey M (1993)
Diffuse and enteroaggregative patterns of adherence of enteric
Escherichia coli isolated from aboriginal children from the
Kimberley region of Western Australia. J Infect Dis 167:
755–758.
Guo H, Feng L, Tao J, Zhang C & Wang L (2004) Identification of
Escherichia coli O172 O-antigen gene cluster and development
of a serogroup-specific PCR assay. J Appl Microbiol 97:
181–190.
Gupta DS, Shashkov AS, Jann B & Jann K (1992) Structures of the
O1B and O1C lipopolysaccharide antigens of Escherichia coli. J
Bacteriol 174: 7963–7970.
Hicks S, Candy DC & Phillips AD (1996) Adhesion of
enteroaggregative Escherichia coli to pediatric intestinal
mucosa in vitro. Infect Immun 64: 4751–4760.
Hoque SS, Faruque AS, Mahalanabis D & Hasnat A (1994)
Infectious agents causing acute watery diarrhoea in infants and
young children in Bangladesh and their public health
implications. J Trop Pediatr 40: 351–354.
Hygge Blackeman K, Weintraub A & Widmalm G (1998)
Structural determination of the O-antigenic polysaccharide
from the enterotoxigenic Escherichia coli O147. Eur J Biochem
251: 534–537.
Jallat C, Livrelli V, Darfeuille-Michaud A, Rich C & Joly B (1993)
Escherichia coli strains involved in diarrhea in France: high
prevalence and heterogeneity of diffusely adhering strains. J
Clin Microbiol 31: 2031–2037.
Jann B, Shashkov AS, Gupta DS & Jann K (1992a) The O18
antigens (lipopolysaccharides) of Escherichia coli. Structural
characterization of the O18A, O18A1, O18B, O18B1-specific
polysaccharides. Eur J Biochem 210: 241–248.
Jann B, Shashkov AS, Gupta DS, Panasenko SM & Jann K (1992b)
The O1 antigen of Escherichia coli: structural characterization
of the O1A1-specific pdysaccharide. Carbohydr Polymers 18:
51–57.
Jann B, Shashkov AS, Kochanowski H & Jann K (1993) Structural
comparison of the O4-specific polysaccharides from E. coli
O4:K6 and E. coli O4:K52. Carbohydr Res 248: 241–250.
Jann B, Shashkov AS, Hahne M, Kochanowski H & Jann K
(1994a) Structure of the O83-specific polysaccharide of
Escherichia coli O83:K24:H31. Carbohydr Res 261: 215–222.
Jann B, Shashkov AS, Kochanowski H & Jann K (1994b) Structure
of the O16 polysaccharide from Escherichia coli O16:K1: an
NMR investigation. Carbohydr Res 264: 305–311.
FEMS Microbiol Rev 30 (2006) 382–403
Jann B, Shashkov AS, Kochanowski H & Jann K (1994c)
Structural comparison of the O6 specific polysaccharides from
E. coli O6:K2:H1, E. coli O6:K13:H1, and E. coli O6:K54:H10.
Carbohydr Res 263: 217–225.
Jann B, Shashkov A, Torgov V, Kochanowski H, Seltmann G &
Jann K (1995) NMR investigation of the 6-deoxy-L-talosecontaining O45, O45-related (O45rel), and O66
polysaccharides of Escherichia coli. Carbohydr Res 278:
155–165.
Jansson PE, Lindberg B, Lönngren J, Ortega C & Svenson SB
(1984) Structural studies of the Escherichia coli O-antigen 6.
Carbohydr Res 131: 277–283.
Jansson PE, Lönngren J, Widmalm G, Leontein K, Slettengren K,
Svenson SB, Wrangsell G, Dell A & Tiller PR (1985) Structural
studies of the O-antigen polysaccharides of Klebsiella O5 and
Escherichia coli O8. Carbohydr Res 145: 59–66.
Jansson PE, Lennholm H, Lindberg B, Lindquist U & Svenson SB
(1987a) Structural studies of the O-specific side-chains of the
Escherichia coli O2 lipopolysaccharide. Carbohydr Res 161:
273–279.
Jansson PE, Lindberg B, Widmalm G & Leontein K (1987b)
Structural studies of the Escherichia coli O78 O-antigen
polysaccharide. Carbohydr Res 165: 87–92.
Jansson PE, Kenne L & Widmalm G (1989) Structure of the Oantigen polysaccharide from Escherichia coli O18ac: a revision
using computer-assisted structural analysis with the program
CASPER. Carbohydr Res 193: 322–325.
Johnson JR (1991) Virulence factors in Escherichia coli urinary
tract infection. Clin Microbiol Rev 4: 80–128.
Kai E, Ikebukuro K, Hoshina S, Watanabe H & Karube I (2000)
Detection of PCR products of Escherichia coli O157:H7 in
human stool samples using surface plasmon resonance (SPR).
FEMS Immunol Med Microbiol 29: 283–288.
Kaper JB (1998) Enterohemorrhagic Escherichia coli. Curr Opin
Microbiol 1: 103–108.
Kaper JB, Nataro JP & Mobley HL (2004) Pathogenic Escherichia
coli. Nat Rev Microbiol 2: 123–140.
Katzenellenbogen E, Kocharova NA, Zatonsky GV, Kübler-Kielb
J, Gamian A, Shashkov AS, Knirel YA & Romanowska E (2001)
Structural and serological studies on Hafnia alvei O-specific
polysaccharide of a-D-mannan type isolated from the
lipopolysaccharide of strain PCM 1223. FEMS Immunol Med
Microbiol 30: 223–227.
Kehl SC (2002) Role of the laboratory in the diagnosis of
enterohemorrhagic Escherichia coli infections. J Clin Microbiol
40: 2711–2715.
Kenne L, Lindberg B, Madden JK, Lindberg AA & Gemski P Jr
(1983a) Structural studies of the Escherichia coli O-antigen 25.
Carbohydr Res 122: 249–256.
Kenne L, Lindberg B, Söderholm E, Bundle DR & Griffith DW
(1983b) Structural studies of the O-antigens from Salmonella
greenside and Salmonella adelaide. Carbohydr Res 111:
289–296.
2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
c
400
Kenne L, Lindberg B, Lugowski C & Svenson SB (1986) Structural
studies of the O-specific side-chains of the Escherichia coli O10
lipopolysaccharide. Carbohydr Res 151: 349–358.
Kerneis S, Bilge SS, Fourel V, Chauviere G, Coconnier MH &
Servin AL (1991) Use of purified F1845 fimbrial adhesin to
study localization and expression of receptors for diffusely
adhering Escherichia coli during enterocytic differentiation of
human colon carcinoma cell lines HT-29 and Caco-2 in
culture. Infect Immun 59: 4013–4018.
Kjellberg A, Urbina F, Weintraub A & Widmalm G (1996)
Structural analysis of the O-antigenic polysaccharide from the
enteropathogenic Escherichia coli O125. Eur J Biochem 239:
532–538.
Kjellberg A, Weintraub A & Widmalm G (1999) Structural
determination and biosynthetic studies of the O-antigenic
polysaccharide from the enterohemorrhagic Escherichia coli
O91 using 13C-enrichment and NMR spectroscopy.
Biochemistry 38: 12205–12211.
Kopecko DJ (1994) Experimental keratoconjunctivitis (Sereny)
assay. Methods Enzymol 235: 39–47.
Landersjö C, Weintraub A & Widmalm G (1996) Structure
determination of the O-antigen polysaccharide from the
enteroinvasive Escherichia coli (EIEC) O143 by component
analysis and NMR spectroscopy. Carbohydr Res 291: 209–216.
Landersjö C, Weintraub A & Widmalm G (1997) Structural
analysis of the O-antigenic polysaccharide from the
enteropathogenic Escherichia coli O142. Eur J Biochem 244:
449–453.
Landersjö C, Weintraub A & Widmalm G (2001) Structural
analysis of the O-antigen polysaccharide from the Shiga toxinproducing Escherichia coli O172. Eur J Biochem 268:
2239–2245.
Larsson EA, Urbina F, Yang Z, Weintraub A & Widmalm G (2004)
Structural and immunochemical relationship between the Oantigenic polysaccharides from the enteroaggregative
Escherichia coli strain 396/C-1 and Escherichia coli O126.
Carbohydr Res 339: 1491–1496.
Lerouge I & Vanderleyden J (2001) O-antigen structural
variation: mechanisms and possible roles in animal/
plant–microbe interactions. FEMS Microbiol Rev 26: 17–47.
Leslie MR, Parolis H & Parolis LA (1999) The structure of the Oantigen of Escherichia coli O116:K1:H10. Carbohydr Res 321:
246–256.
Leslie MR, Parolis H & Parolis LA (2000) The structure of the Ospecific polysaccharide of Escherichia coli O117:K98:H4.
Carbohydr Res 323: 103–110.
Levine MM, Caplan ES, Waterman D, Cash RA, Hornick RB &
Snyder MJ (1977) Diarrhea caused by Escherichia coli that
produce only heat-stable enterotoxin. Infect Immun 17: 78–82.
Levine MM & Edelman R (1984) Enteropathogenic Escherichia
coli of classic serotypes associated with infant diarrhea:
epidemiology and pathogenesis. Epidemiol Rev 6: 31–51.
Levine MM (1987) Escherichia coli that cause diarrhea:
enterotoxigenic, enteropathogenic, enteroinvasive,
2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
c
R. Stenutz et al.
enterohemorrhagic, and enteroadherent. J Infect Dis 155:
377–389.
Levine MM, Ferreccio C, Prado V, et al. (1993) Epidemiologic
studies of Escherichia coli diarrheal infections in a low
socioeconomic level peri-urban community in Santiago, Chile.
Am J Epidemiol 138: 849–869.
Lima AA, Fang G, Schorling JB, de Albuquerque L, Mc Auliffe JF,
Mota S, Leite R & Guerrant RL (1992) Persistent diarrhea in
northeast Brazil: etiologies and interactions with malnutrition.
Acta Paediatr Suppl 381: 39–44.
Lindberg B, Lindh F & Lönngren J (1981) Structural studies of the
O-specific side-chain of the lipopolysaccharide from
Escherichia coli O55. Carbohydr Res 97: 105–112.
Linnerborg M, Weintraub A & Widmalm G (1997a) Structural
studies of the O-antigen polysaccharide from Escherichia coli
O138. Eur J Biochem 247: 567–571.
Linnerborg M, Wollin R & Widmalm G (1997b) Structural
studies of the O-antigenic polysaccharide from Escherichia coli
O167. Eur J Biochem 246: 565–573.
Linnerborg M, Weintraub A & Widmalm G (1999a) Structural
studies of the O-antigen polysaccharide from the
enteroinvasive Escherichia coli O164 cross-reacting with
Shigella dysenteriae type 3. Eur J Biochem 266: 460–466.
Linnerborg M, Weintraub A & Widmalm G (1999b) Structural
studies of the O-antigen polysaccharide from the
enteroinvasive Escherichia coli O173. Carbohydr Res 320:
200–208.
Linnerborg M, Weintraub A & Widmalm G (1999c) Structural
studies utilizing 13C-enrichment of the O-antigen
polysaccharide from the enterotoxigenic Escherichia coli O159
cross-reacting with Shigella dysenteriae type 4. Eur J Biochem
266: 246–251.
Liu D & Reeves PR (1994) Escherichia coli K12 regains its O
antigen. Microbiology 140: 49–57.
L’vov VL, Shashkov AS, Dmitriev BA, Kochetkov NK, Jann B &
Jann K (1984) Structural studies of the O-specific side chain of
the lipopolysaccharide from Escherichia coli O:7. Carbohydr
Res 126: 249–259.
L’vov VL, Iakovlev AP, Shashkov AS & Dmitriev BA (1991)
Antigenic polysaccharides of Shigella bacteria. Structure of the
polysaccharide chain of the lipopolysaccharide from Shigella
boydii, type 11. Bioorg Khim 17: 111–120.
Lycknert K & Widmalm G (2004) Dynamics of the Escherichia coli
O91 O-antigen polysaccharide in solution as studied by
carbon-13 NMR relaxation. Biomacromolecules 5: 1015–1020.
MacLean LL & Perry MB (1997) Structural characterization of
the serotype O:5 O-polysaccharide antigen of the
lipopolysaccharide of Escherichia coli O:5. Biochem Cell Biol 75:
199–205.
Manca MC, Weintraub A & Widmalm G (1996) Structural studies
of the Escherichia coli O26 O-antigen polysaccharide.
Carbohydr Res 281: 155–160.
Manges AR, Johnson JR, Foxman B, O’Bryan TT, Fullerton KE &
Riley LW (2001) Widespread distribution of urinary tract
FEMS Microbiol Rev 30 (2006) 382–403
401
Escherichia coli O-polysaccharide antigens
infections caused by a multidrug-resistant Escherichia coli
clonal group. N Engl J Med 345: 1007–1013.
Mangia AH, Duarte AN, Duarte R, Silva LA, Bravo VL & Leal MC
(1993) Aetiology of acute diarrhoea in hospitalized children in
Rio de Janeiro City, Brazil. J Trop Pediatr 39: 365–367.
Marie C, Weintraub A & Widmalm G (1998) Structural studies of
the O-antigenic polysaccharide from Escherichia coli O139. Eur
J Biochem 254: 378–381.
Marolda CL, Vicarioli J & Valvano MA (2004) Wzx proteins
involved in biosynthesis of O antigen function in association
with the first sugar of the O-specific lipopolysaccharide
subunit. Microbiology 150: 4095–4105.
Marsden BJ, Bundle DR & Perry MB (1994) Serological and
structural relationships between Escherichia coli O:98 and
Yersinia enterocolitica O:11,23 and O:11,24 lipopolysaccharide
O-antigens. Biochem Cell Biol 72: 163–168.
Masoud H & Perry MB (1996) Structural characterization of the
O-antigenic polysaccharide of Escherichia coli serotype O17
lipopolysaccharide. Biochem Cell Biol 74: 241–248.
McConnell M.M., Smith H.R., Willshaw G.A., Field A.M. & Rowe
B. (1981) Plasmids coding for colonization factor antigen I and
heat-stable enterotoxin production isolated from
enterotoxigenic Escherichia coli: comparison of their
properties. Infect Immun 32: 927–936.
Medina EC, Widmalm G, Weintraub A, Vial PA, Levine MM &
Lindberg AA (1994) Structural studies of the O-antigenic
polysaccharides of Escherichia coli O3 and the
enteroaggregative Escherichia coli strain 17-2. Eur J Biochem
224: 191–196.
Moran AP, Prendergast MM & Appelmelk BJ (1996) Molecular
mimicry of host structures by bacterial lipopolysaccharides
and its contribution to disease. FEMS Immunol Med Microbiol
16: 105–115.
Murray BE, Evans DJ Jr, Penaranda ME & Evans DG (1983) CFA/
I-heat-stable plasmids: comparison of enterotoxigenic
Escherichia coli (ETEC) of serogroups O25, O63, O78, and
O128 and mobilization from an R factor-containing epidemic
ETEC isolate. J Bacteriol 153: 566–570.
Nataro JP & Kaper JB (1998) Diarrheagenic Escherichia coli. Clin
Microbiol Rev 11: 142–201.
Nataro JP, Kaper JB, Robins-Browne R, Prado V, Vial P & Levine
MM (1987) Patterns of adherence of diarrheagenic Escherichia
coli to HEp-2 cells. Pediatr Infect Dis J 6: 829–831.
Nataro JP, Steiner T & Guerrant RL (1998) Enteroaggregative
Escherichia coli. Emerg Infect Dis 4: 251–261.
Nishiuchi Y, Doe M, Hotta H & Kobayashi K (2000) Structure
and serologic properties of O-specific polysaccharide from
Citrobacter freundii possessing cross-reactivity with Escherichia
coli O157:H7. FEMS Immunol Med Microbiol 28: 163–171.
Nishiuchi Y, Doe M, Hotta H & Kobayashi K (2002) Addendum
to: ‘‘Structure and serological properties of O-specific
polysaccharide from Citrobacter freundii possessing crossreactivity with Escherichia coli O157:H7’’ [FEMS Immunol.
Med. Microbiol. 28 (2000) 163–171]. Med. Microbiol 28:
163–171].
FEMS Microbiol Rev 30 (2006) 382–403
Nowicki B, Svanborg-Eden C, Hull R & Hull S (1989) Molecular
analysis and epidemiology of the Dr hemagglutinin of
uropathogenic Escherichia coli. Infect Immun 57: 446–451.
Olsson U, Lycknert K, Stenutz R, Weintraub A & Widmalm G
(2005) Structural analysis of the O-antigen polysaccharide
from Escherichia coli O152. Carbohydr Res 340: 167–171.
Orskov F & Orskov I (1984) Serotyping of Escherichia coli.
Methods in Microbiology, Vol. 14 (Bergan T, ed.), pp. 43–112.
Academic Press Inc., London.
Oxley D & Wilkinson SG (1986) Structure of the O-specific
polysaccharide from the lipopolysaccharide of Serratia
marcescens O8. Eur J Biochem 156: 597–601.
Ozeki Y, Kurazono T, Saito A, Kishimoto T & Yamaguchi M
(2003) A diffuse outbreak of enterohemorrhagic Escherichia
coli O157:H7 related to the Japanese-style pickles in Saitama,
Japan. Kansenshogaku Zasshi 77: 493–498.
Parolis H & Parolis LA (1995) The structure of the O-specific
polysaccharide from Escherichia coli O113 lipopolysaccharide.
Carbohydr Res 267: 263–269.
Parolis LA, Parolis H & Dutton GG (1986) Structural studies of
the O-antigen polysaccharide of Escherichia coli O9a.
Carbohydr Res 155: 272–276.
Parolis H, Parolis LA & Olivieri G (1997) Structural studies on the
Shigella-like Escherichia coli O121 O-specific polysaccharide.
Carbohydr Res 303: 319–325.
Parsot C & Sansonetti PJ (1996) Invasion and the pathogenesis of
Shigella infections. Curr Top Microbiol Immunol 209: 25–42.
Penaranda ME, Evans DG, Murray BE & Evans DJ, Jr. (1983)
ST:LT:CFA/II plasmids in enterotoxigenic Escherichia coli
belonging to serogroups O6, O8, O80, O85, and O139. J
Bacteriol 154: 980–983.
Perry MB & Mac Lean LL (1987) Structure of the
lipopolysaccharide O-chain of Yersinia enterocolitica serotype
O:5,27. Biochem Cell Biol 65: 1–7.
Perry MB & Mac Lean LL (1999) Structural characterization of
the antigenic O-chain of the lipopolysaccharide of Escherichia
coli serotype O65. Carbohydr Res 322: 57–66.
Perry MB, Mac Lean LL & Brisson JR (1993) The characterization
of the O-antigen of Escherichia coli O64:K99
lipopolysaccharide. Carbohydr Res 248: 277–284.
Perry MB, Mac Lean L & Griffith DW (1986) Structure of the Ochain polysaccharide of the phenol-phase soluble
lipopolysaccharide of Escherichia coli O:157:H7. Biochem Cell
Biol 64: 21–28.
Phillips I, Eykyn S, King A, Gransden WR, Rowe B, Frost JA &
Gross RJ (1988) Epidemic multiresistant Escherichia coli
infection in West Lambeth Health District. Lancet 1:
1038–1041.
Poitrineau P, Forestier C, Meyer M, Jallat C, Rich C, Malpuech G
& De Champs C (1995) Retrospective case-control study of
diffusely adhering Escherichia coli and clinical features in
children with diarrhea. J Clin Microbiol 33: 1961–1962.
Prehm P, Jann B & Jann K (1976) The O9 antigen of Escherichia
coli. Structure of the polysaccharide chain. Eur J Biochem 67:
53–56.
2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
c
402
Pulz M, Matussek A, Monazahian M, Tittel A, Nikolic E,
Hartmann M, Bellin T, Buer J & Gunzer F (2003) Comparison
of a shiga toxin enzyme-linked immunosorbent assay and two
types of PCR for detection of shiga toxin-producing
Escherichia coli in human stool specimens. J Clin Microbiol 41:
4671–4675.
Raetz CR & Whitfield C (2002) Lipopolysaccharide endotoxins.
Annu Rev Biochem 71: 635–700.
Ramotar K, Waldhart B, Church D, Szumski R & Louie TJ (1995)
Direct detection of verotoxin-producing Escherichia coli in
stool samples by PCR. J Clin Microbiol 33: 519–524.
Ratnayake S, Weintraub A & Widmalm G (1994a) Structural
studies of the enterotoxigenic Escherichia coli (ETEC) O153
O-antigenic polysaccharide. Carbohydr Res 265: 113–120.
Ratnayake S, Widmalm G, Weintraub A & Medina EC (1994b)
Structural studies of the Escherichia coli O90 O-antigen
polysaccharide. Carbohydr Res 263: 209–215.
Reis MH, Heloiza M, Affonso T, Trabulsi LR, Mazaitis AJ, Maas R
& Maas WK (1980) Transfer of a CFA/I-heat-stable plasmid
promoted by a conjugative plasmid in a strain of Escherichia
coli of serotype O128ac:H12. Infect Immun 29: 140–143.
Robins-Browne RM & Hartland EL (2002) Escherichia coli as a
cause of diarrhea. J Gastroenterol Hepatol 17: 467–475.
Robins-Browne RM, Still CS, Miliotis MD, Richardson NJ,
Koornhof HJ, Freiman I, Schoub BD, Lecatsas G & Hartman E
(1980) Summer diarrhoea in African infants and children.
Arch Dis Child 55: 923–928.
Rundlöf T, Weintraub A & Widmalm G (1996) Structural studies
of the enteroinvasive Escherichia coli (EIEC) O28 O-antigenic
polysaccharide. Carbohydr Res 291: 127–139.
Rundlöf T, Weintraub A & Widmalm G (1998) Structural
determination of the O-antigenic polysaccharide from
Escherichia coli O35 and cross-reactivity to Salmonella arizonae
O62. Eur J Biochem 258: 139–143.
Samuel B (1996) Medical Microbiology, pp. 303–310). The
University of Texas Medical Branch at Galveston, Texas.
Samuel G & Reeves P (2003) Biosynthesis of O-antigens: genes
and pathways involved in nucleotide sugar precursor synthesis
and O-antigen assembly. Carbohydr Res 338: 2503–2519.
Sasakawa C, Buysse JM & Watanabe H (1992) The large virulence
plasmid of Shigella. Curr Top Microbiol Immunol 180: 21–44.
Scaletsky IC, Pedroso MZ & Fagundes-Neto U (1996) Attaching
and effacing enteropathogenic Escherichia coli o18ab invades
epithelial cells and causes persistent diarrhea. Infect Immun 64:
4876–4881.
Scaletsky IC, Fabbricotti SH, Carvalho RL, Nunes CR, Maranhao
HS, Morais MB & Fagundes-Neto U (2002) Diffusely adherent
Escherichia coli as a cause of acute diarrhea in young children
in Northeast Brazil: a case–control study. J Clin Microbiol 40:
645–648.
Scheutz F (2004) Personal communication.
Scheutz F, Cheasty T, Woodward D & Smith HR (2004)
Designation of O174 and O175 to temporary O groups OX3
and OX7, and six new E. coli O groups that include
2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
c
R. Stenutz et al.
Verocytotoxin-producing E. coli (VTEC): O176, O177, O178,
O179, O180 and O181. Apmis 112: 569–584.
Schultsz C, Pool GJ, van Ketel R, de Wever B, Speelman P &
Dankert J (1994) Detection of enterotoxigenic Escherichia coli
in stool samples by using nonradioactively labeled
oligonucleotide DNA probes and PCR. J Clin Microbiol 32:
2393–2397.
Sengupta P, Bhattacharyya T, Shashkov AS, Kochanowski H &
Basu S (1995) Structure of the O-specific side chain of the
Escherichia coli O128 lipopolysaccharide. Carbohydr Res 277:
283–290.
Sixma TK, Kalk KH, van Zanten BA, Dauter Z, Kingma J, Witholt
B & Hol WG (1993) Refined structure of Escherichia coli heatlabile enterotoxin, a close relative of cholera toxin. J Mol Biol
230: 890–918.
Small PL & Falkow S (1988) Identification of regions on a 230kilobase plasmid from enteroinvasive Escherichia coli that are
required for entry into HEp-2 cells. Infect Immun 56: 225–229.
Smith HR, Scotland SM & Rowe B (1983) Plasmids that code for
production of colonization factor antigen II and enterotoxin
production in strains of Escherichia coli. Infect Immun 40:
1236–1239.
Snyder JD, Wells JG, Yashuk J, Puhr N & Blake PA (1984)
Outbreak of invasive Escherichia coli gastroenteritis on a cruise
ship. Am J Trop Med Hyg 33: 281–284.
Staaf M, Urbina F, Weintraub A & Widmalm G (1997) Structure
determination of the O-antigenic polysaccharide from the
enterotoxigenic Escherichia coli (ETEC) O101. Carbohydr Res
297: 297–299.
Staaf M, Widmalm G, Weintraub A & Nataro JP (1995) Structural
elucidation of the O-antigenic polysaccharide from Escherichia
coli O44:H18. Eur J Biochem 233: 473–477.
Staaf M, Urbina F, Weintraub A & Widmalm G (1999a) Structural
elucidation of the O-antigenic polysaccharides from
Escherichia coli O21 and the enteroaggregative Escherichia coli
strain 105. Eur J Biochem 266: 241–245.
Staaf M, Urbina F, Weintraub A & Widmalm G (1999b) Structure
elucidation of the O-antigenic polysaccharide from the
enteroaggregative Escherichia coli strain 62D1. Eur J Biochem
262: 56–62.
Staaf M, Weintraub A & Widmalm G (1999c) Structure
determination of the O-antigenic polysaccharide from the
enteroinvasive Escherichia coli O136. Eur J Biochem 263:
656–661.
Stacy-Phipps S, Mecca JJ & Weiss JB (1995) Multiplex PCR assay
and simple preparation method for stool specimens detect
enterotoxigenic Escherichia coli DNA during course of
infection. J Clin Microbiol 33: 1054–1059.
Stevenson G, Neal B, Liu D, Hobbs M, Packer NH, Batley M,
Redmond JW, Lindquist L & Reeves P (1994) Structure of the
O antigen of Escherichia coli K-12 and the sequence of its rfb
gene cluster. J Bacteriol 176: 4144–4156.
Tao J, Feng L, Guo H, Li Y & Wang L (2004) The O-antigen gene
cluster of Shigella boydii O11 and functional identification of
its wzy gene. FEMS Microbiol Lett 234: 125–132.
FEMS Microbiol Rev 30 (2006) 382–403
403
Escherichia coli O-polysaccharide antigens
Taylor DN, Echeverria P, Sethabutr O, Pitarangsi C, Leksomboon
U, Blacklow NR, Rowe B, Gross R & Cross J (1988) Clinical
and microbiologic features of Shigella and enteroinvasive
Escherichia coli infections detected by DNA hybridization. J
Clin Microbiol 26: 1362–1366.
Thomas LV, Rowe B & Mc Connell MM (1987) In strains of
Escherichia coli O167 a single plasmid encodes for the coli
surface antigens CS5 and CS6 of putative colonization factor
PCF8775, heat-stable enterotoxin, and colicin Ia. Infect Immun
55: 1929–1931.
Tomasz A (ed) (2000) Streptococcus pneumoniae: Molecular
Biology and Mechanisms of Disease. Mary Ann Liebert, New
York.
Torgov VI, Shashkov AS, Jann B & Jann K (1995) NMR
reinvestigation of two N-acetylneuraminic acid-containing Ospecific polysaccharides (O56 and O24) of Escherichia coli.
Carbohydr Res 272: 73–90.
Torgov VI, Shashkov AS, Kochanowski H, Jann B & Jann K (1996)
NMR analysis of the structure of the O88 polysaccharide (O88
antigen) of Escherichia coli O88:K:H25. Carbohydr Res 283:
223–227.
Unhanand M, Mustafa MM, Mc Cracken GHJr & Nelson JD
(1993) Gram-negative enteric bacillary meningitis: a twentyone-year experience. J Pediatr 122: 15–21.
Urbina F, Nordmark E-L, Yang Z, Weintraub A, Scheutz F &
Widmalm G (2005) Structural elucidation of the O-antigen
polysaccharide from the enteroaggregative Escherichia coli
strain 180/C3 and its immunochemical relationship with E.
coli O5 and O65. Carbohydr Res 340: 645–650.
Varki A, Cummings R, Esko J, Freeze H, Hart G & Marth J (1999)
Essentials of Glycobiology. Cold Spring Harbor Laboratory
Press, New York.
FEMS Microbiol Rev 30 (2006) 382–403
Vasil’ev VN & Zakharova IY (1976) Structure of the determinant
group of the O-specific polysaccharide of E. coli O20:K84:H34
(145). Bioorg Khim 2: 199–206.
Vasil’ev VN & Zakharova IY (1982) Structure of the O-specific
polysaccharides of Escherichia coli Str. O20ab:K84:H34 (145)
and O20ac:K61:H according to 13C NMR spectroscopy.
Bioorg Khim 8: 120–125.
Vial PA, Mathewson JJ, DuPont HL, Guers L & Levine MM
(1990) Comparison of two assay methods for patterns of
adherence to HEp-2 cells of Escherichia coli from patients with
diarrhea. J Clin Microbiol 28: 882–885.
Vinogradov E, Conlan JW & Perry MB (2000) Serological crossreaction between the lipopolysaccharide O-polysaccharaide
antigens of Escherichia coli O157:H7 and strains of Citrobacter
freundii and Citrobacter sedlakii. FEMS Microbiol Lett 190:
157–161.
Weintraub A, Leontein K, Widmalm G, Vial PA, Levine MM &
Lindberg AA (1993) Structural studies of the O-antigenic
polysaccharide of an enteroaggregative Escherichia coli strain.
Eur J Biochem 213: 859–864.
Whitfield C & Roberts IS (1999) Structure, assembly and
regulation of expression of capsules in Escherichia coli. Mol
Microbiol 31: 1307–1319.
Widmalm G & Leontein K (1993) Structural studies of the
Escherichia coli O127 O-antigen polysaccharide. Carbohydr Res
247: 255–262.
Wolf MK (1997) Occurrence, distribution, and associations of O
and H serogroups, colonization factor antigens, and toxins of
enterotoxigenic Escherichia coli. Clin Microbiol Rev 10:
569–584.
Yildirim H, Weintraub A & Widmalm G (2001) Structural studies
of the O-polysaccharide from the Escherichia coli O77
lipopolysaccharide. Carbohydr Res 333: 179–183.
2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
c