Monoclonal antibodies as probes for detecting lipopolysaccharide expression on Escherichia coli from different growth conditions

Nelson, David Lee; Bathgate, Andrew; Poxton, Ian R.

doi:10.1099/00221287-137-12-2741

Cited by 26 publications

(12 citation statements)

References 34 publications

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“…Core epitopes are, however, much more accessible on bacterial cells grown in serum or in a magnesium-depleted medium, which is thought to mimic the conditions in the infected host [20]. Core epitopes also become more accessible in bacterial preparations heated at 100°C [21,22].…”

Section: Discussionmentioning

confidence: 99%

Frequencies of Lipopolysaccharide Core Types among Clinical Isolates of Escherichia coli Defined with Monoclonal Antibodies

Gibb¹,

Barclay²,

Poxton³

et al. 1992

Journal of Infectious Diseases

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Mouse monoclonal antibodies (MAbs) specific for the lipopolysaccharide (LPS) core types R1, R2, and R3 of Escherichia coliand a cross-reactive MAb that binds to the LPS core of almost all E. coli were used in ELISA to determine the frequency of cores resembling R1, R2, and R3 in strains of E. coli isolated from clinical samples (blood and urine specimens) and from the feces of asymptomatic individuals. Of the 180 wild-type isolates, 123 were assigned to R1 core type, 14 to R2, and 18 to R3. Twenty-five wild-type E. coliisolates could not be assigned to a particular core type and may have either an R4 or K12 core or a previously unrecognized core type. R1 core type was associated with 0 types 1,4,6,8, and 18 and with K1 or K5 capsules. R3 was associated with 015. 075 isolates could be of either R1 or R2 core type.The lipopolysaccharide (LPS) core region of Escherichia coli has a conserved overall structure, with an inner 2-keto-3-deoxy-octonate-heptose region and an outer hexose region. Most of the variation that does occur is in the outer part of the core, furthest from lipid A. In wild-type smooth E. coli. the core region can occur in at least four different forms, known as R 1, R2, R3, and R4 [1][2][3]. The naturally rough strain Kl2 has a different core region [4].The 0 antigen of smooth LPS may mask some of the epitopes and phage attachment sites in the core region. The chemical, serologic, and phage typing methods that have been used to define the different core types of E. coli are therefore not directly applicable to wild-type smooth strains. The frequency with which these different core types occur in wild-type E. coli is therefore not known.Little is known about the association between core types and 0 serotypes ofE. coli. except that the original R 1 and R2 rough mutants were both derived from 08 strains [I] and that the original R3 mutant was derived from an 0 III strain [2]. There is no published evidence on the relationship of LPS core types to capsular types or to virulence. Information about the distribution of 0 and K antigens has proved to be useful, for example in identifying and understanding enteropathogenic strains [5] and in suggesting novel therapeutic approaches to bacteremia [6]. Information about core types might be equally interesting.We therefore set out to produce a panel ofmouse monoclo- Materials and MethodsBacteria and bacteriophages. E. coli RI (F470), R2 (F576), R3 (F653), R4 (2513) and KI2 (2131) and rough-specific phages FO, BrIO, C21, 6SR, and T4 were obtained from G. Schmidt (Forschunginstitut, Borstel, Germany). E. coli 0 I, 02, 04, 06, 012, 015, 018, and 075 were obtained from A. S. Cross (Walter Reed Army Institute, Washington, DC). Phages specific for E. coli capsular types KI (KIGS) and K5 (K5DG) were obtained from A. P. Roberts (Charing Cross and Westminster Medical School, London). Blood culture and urine isolates were obtained from the Clinical Bacteriology Laboratory, Department of Medical Microbiology, Edinburgh University, from routine clinical specimens. ...

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Section: Discussionmentioning

confidence: 99%

Frequencies of Lipopolysaccharide Core Types among Clinical Isolates of Escherichia coli Defined with Monoclonal Antibodies

Gibb¹,

Barclay²,

Poxton³

et al. 1992

Journal of Infectious Diseases

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“…This has been largely supported by recent work by Amor et al [11], who showed by PCR that the great majority of verocytotoxigenic E. coli (VTEC) strains possessed the R3 core. The complete core oligosaccharide-lipid A (R-form LPS) is naturally expressed on wild-type smooth bacteria and is accessible to antibody; it is visualised as the bottom`rung' of the ladder pattern in SDS-PAGE [12].…”

Section: Introductionmentioning

confidence: 99%

Mucosal and systemic antibody responses to the lipopolysaccharide of Escherichia coli O157 in health and disease*

Currie

McCALLUM

Poxton

2001

Journal of Medical Microbiology

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Mucosal immunity in the gastrointestinal (GI) tract is a primary defence against GI pathogens. We hypothesise that a mucosal response to lipopolysaccharide (LPS), especially to the common (core) determinants of GI pathogenic Escherichia coli strains, is protective. The aims of this study were to investigate the speci®cities, levels and development of humoral responses in health and GI disease to the R3 LPS core and O-polysaccharide of E. coli O157. The purpose was to try to predict whether vaccination or passive immunisation might induce protection. Wherever possible, paired whole gut lavage¯uid (WGLF) and serum samples were collected for comparison of the mucosal and systemic responses. Matched saliva samples were also collected from some study groups. The patient groups included those with acute E. coli O157 disease (serum only), patients convalescing after E. coli O157 infections, and patients undergoing routine investigation for GI conditions but subsequently shown to be immunologically normal. Some samples of WGLF from patients with Crohn's disease (CRO) and ulcerative colitis (UC) were included to allow comparisons with patients with in¯ammatory conditions known to alter antibody secretion in the GI tract. The healthy groups from whom serum and saliva only were taken included blood donors, healthy volunteers and a group of slaughterhouse workers. This latter group was likely to have been exposed regularly to faecal bacteria from animals and antibody speci®cities might have been expected to be different from other healthy individuals. Levels and classes of antibodies were determined by ELISA with microtitration plates coated with polymyxin complexes of whole LPS extracted from E. coli O157 and LPS from the E. coli R3 rough mutant. Antibodies of IgG and IgM classes were measured in serum and IgA was measured in WGLF and saliva. IgG antibodies to the O157 LPS and the R3 core oligosaccharide were detected in the serum of healthy blood donors. Patients with acute E. coli O157 disease showed elevated levels of serum IgM to O157 LPS and R3, with IgG levels raised only to R3. In serum from convalescent patients, IgG to O157 LPS was signi®cantly above the control groups only in the period 6±16 weeks after infection. Total IgA levels were similar in WGLF specimens from all groups, except the patients with UC, whose levels were much higher. Speci®c IgA levels were higher in the E. coli O157 convalescent group, but there were no signi®cant correlations overall. UC patients had signi®cantly lower levels of IgA to O157 and CRO patients had higher O157 IgA levels than UC patients and healthy volunteers. In serum, inhibition of ELISA showed that the response to the O157 LPS was due in part to a response to the R3 oligosaccharide component. This response was much more pronounced in the healthy and non-O157 groups than in convalescent patients. There was no correlation between speci®c IgA antibody levels in saliva and matched specimens of WGLF, and levels in sequential saliva specimens¯uctuated widely. The signi®cant IgG an...

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“…One limitation to using monoclonal antibodies to classify soil streptomycetes is that the lownutrient environment may cause changes in the accessibility and expression of surface antigens, which might affect antibody binding. This has been demonstrated for Gram-negative micro-organisms (Nelson et al, 1991) and there is limited evidence that it may also occur with Gram-positive micro-organisms (Smith et al, 1991). …”

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confidence: 96%

The taxonomy of Streptomyces and related genera.

Anderson¹,

Wellington²

2001

International Journal of Systematic and Evolutionary Microbiology

308

174

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The streptomycetes, producers of more than half of the 10 000 documented bioactive compounds, have offered over 50 years of interest to industry and academia. Despite this, their taxonomy remains somewhat confused and the definition of species is unresolved due to the variety of morphological, cultural, physiological and biochemical characteristics that are observed at both the inter-and the intraspecies level. This review addresses the current status of streptomycete taxonomy, highlighting the value of a polyphasic approach that utilizes genotypic and phenotypic traits for the delimitation of species within the genus.Keywords : streptomycete taxonomy, phylogeny, numerical taxonomy, fingerprinting, bacterial systematics IntroductionThe genus Streptomyces was proposed by Waksman & Henrici (1943) and classified in the family Streptomycetaceae on the basis of morphology and subsequently cell wall chemotype. The development of numerical taxonomic systems, which utilized phenotypic traits helped to resolve the intergeneric relationships within the family Streptomycetaceae and resulted in the reclassification of six additional genera (Actinopycnidium, Actinosporangium, Chainia, Elytrosporangium, Kitasatoa and Microellobosporia) to the Streptomyces genus (Williams et al., 1983a ; Goodfellow et al., 1986a-d). These early numerical systems utilized phenotypic characters, which were fundamentally changed by the incorporation of molecular biological characteristics into classification systems and thus enabled considerable advances for genus delimitation within the Actinobacteria (Stackebrandt et al., 1997). Prior to this, the genera Streptomyces and Streptoverticillium were two distinct genera ; both have cell-wall type 1 (Lechevalier & Lechevalier, 1970), are lysed by the same phages (Wellington & Williams, 1981) and are phylogenetically closely related (Stackebrandt & Woese, 1981). Immunodiffusion studies (Ridell et al., 1986) linked members of the genus Streptoverticillium closely to the Streptomyces lavendulae species group. Ka$ mpfer et al. (1991) also found similarities using physiological tests. Gladek et al. (1985) observed differences in DNA-RNA pairing ; this and the morphological trait of producing whorls were the only detectable differences between the two genera. Witt & Stackebrandt (1990) concluded from 16S and 23S rRNA comparisons that the genus Streptoverticillium should be regarded as a synonym of Streptomyces. Kitasatosporia was also included in the genus Streptomyces, despite having differences in cell wall composition, on the basis of 16S rRNA similarities (Wellington et al., 1992). This was revoked by Zhang et al. (1997), who demonstrated that members of the genus Kitasatosporia always formed a stable monophyletic clade away from streptomycetes when sequences from the entire 16S rRNA genes were compared. Kineosporia and Sporichthya are both rare and share many chemotaxonomic similarities with members of the genus Streptomyces, which led to their incorporation into the genus (Logan, 1994). The Kine...

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Monoclonal antibodies as probes for detecting lipopolysaccharide expression on Escherichia coli from different growth conditions

Cited by 26 publications

References 34 publications

Frequencies of Lipopolysaccharide Core Types among Clinical Isolates of Escherichia coli Defined with Monoclonal Antibodies

Frequencies of Lipopolysaccharide Core Types among Clinical Isolates of Escherichia coli Defined with Monoclonal Antibodies

Mucosal and systemic antibody responses to the lipopolysaccharide of Escherichia coli O157 in health and disease*

The taxonomy of Streptomyces and related genera.

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