Representative Shigella flexneri strains were studied to determine whether plasmids are involved in their virulence. All invasive S. flexneri strains, irrespective of serotype, were found to harbor a large plasmid of approximately 140 megadaltons in size, although some strains carried additional plasmid species. Spontaneous variants of strains of serotypes 1, 2, and 5 had lost this 140-megadalton plasmid and had concomitantly become avirulent, i.e., could neither invade HeLa cell monolayers nor produced keratoconjunctivitis in guinea pigs. To monitor plasmid transfer, the 140-megadalton plasmid of strain M90T (serotype 5) was tagged with the kanamycin resistance transposon Tn5. This tagged plasmid, pWR110, was not self-transmissible, but was mobilized by one of several different conjugative plasmids into avirulent derivatives of the heterologous serotypes 1 and 2 which had lost the comparable large plasmid. Transconjugants of both serotypes which had received pWR110 regained virulence. These data directly demonstrate that this large S. flexneri plasmid encodes or regulates some function(s) required for epithelial cell penetration.

Involvement of a plasmid in the invasive ability of Shigella flexneri.

Sialic acids and their role as biological masks

Structure of bacterial lipopolysaccharides.

Two rat monoclonal antibodies have been generated to plant cell wall (1→4)-β-D-xylans using a penta-1,4-xylanoside-containing neoglycoprotein as an immunogen. The monoclonal antibodies, designated LM10 and LM11, have different specificities to xylans in relation to the substitution of the xylan backbone as indicated by immunodot assays and competitive-inhibition ELISAs. LM10 is specific to unsubstituted or low-substituted xylans, whereas LM11 binds to wheat arabinoxylan in addition to unsubstituted xylans. Immunocytochemical analyses indicated the presence of both epitopes in secondary cell walls of xylem but differences in occurrence in other cell types.

Monoclonal antibodies to plant cell wall xylans and arabinoxylans.

Current data from bacterial pathogens of animals and from bacterial symbionts of plants support some of the more general proposed functions for lipopolysaccharides (LPS) and underline the importance of LPS structural versatility and adaptability. Most of the structural heterogeneity of LPS molecules is found in the O-antigen polysaccharide. In this review, the role and mechanisms of this striking flexibility in molecular structure of the O-antigen in bacterial pathogens and symbionts are illustrated by some recent findings. The variation in O-antigen that gives rise to an enormous structural diversity of O-antigens lies in the sugar composition and the linkages between monosaccharides. The chemical composition and structure of the O-antigen is strain-specific (interstrain LPS heterogeneity) but can also vary within one bacterial strain (intrastrain LPS heterogeneity). Both LPS heterogeneities can be achieved through variations at different levels. First of all, O-polysaccharides can be modified non-stoichiometrically with sugar moieties, such as glucosyl and fucosyl residues. The addition of non-carbohydrate substituents, i.e. acetyl or methyl groups, to the O-antigen can also occur with regularity, but in most cases these modifications are again non-stoichiometric. Understanding LPS structural variation in bacterial pathogens is important because several studies have indicated that the composition or size of the O-antigen might be a reliable indicator of virulence potential and that these important features often differ within the same bacterial strain. In general, O-antigen modifications seem to play an important role at several (at least two) stages of the infection process, including the colonization (adherence) step and the ability to bypass or overcome host defense mechanisms. There are many reports of modifications of O-antigen in bacterial pathogens, resulting either from altered gene expression, from lysogenic conversion or from lateral gene transfer followed by recombination. In most cases, the mechanisms underlying these changes have not been resolved. However, in recent studies some progress in understanding has been made. Changes in O-antigen structure mediated by lateral gene transfer, O-antigen conversion and phase variation, including fucosylation, glucosylation, acetylation and changes in O-antigen size, will be discussed. In addition to the observed LPS heterogeneity in bacterial pathogens, the structure of LPS is also altered in bacterial symbionts in response to signals from the plant during symbiosis. It appears to be part of a molecular communication between bacterium and host plant. Experiments ex planta suggest that the bacterium in the rhizosphere prepares its LPS for its roles in symbiosis by refining the LPS structure in response to seed and root compounds and the lower pH at the root surface. Moreover, modifications in LPS induced by conditions associated with infection are another indication that specific structures are important. Also during the differentiation from bacterium to bacteroid, the LPS of Rhizobium undergoes changes in the composition of the O-antigen, presumably in response to the change of environment. Recent findings suggest that, during symbiotic bacteroid development, reduced oxygen tension induces structural modifications in LPS that cause a switch from predominantly hydrophilic to predominantly hydrophobic molecular forms. However, the genetic mechanisms by which the LPS epitope changes are regulated remain unclear. Finally, the possible roles of O-antigen variations in symbiosis will be discussed.

http://dzumenvis.nic.in/Microbes%20and%20Metals%20Interaction/pdf/O-antigen%20structural%20variation.pdf

O-antigen structural variation: mechanisms and possible roles in animal/plant-microbe interactions.

The structures of the O-antigens of all known serotypes and subserotypes of Shigella flexneri have been reinvestigated. The results support the assumption that these antigens are composed of a basic tetrasaccharide repeating unit (1), to which alpha-D-glucopyranosyl and/or O-acetyl groups are attached at different positions. Leads to 3)-beta-D-GlcNAcp-(1 leads to 2)-alpha-L-Rhap-(1 leads to 2)-alpha-L-Rhap-(1 leads to 3)-alpha-L-Rhap-(1 leads to The immunological determinants responsible for O-factors I, II, IV, V and 7, 8 contain alpha-D-gluco-pyranosyl groups, the locations of which have been determined. O-Factor 6 is due to O-acetyl groups, linked to O-2 of the 3-substituted alpha-L-rhamnopyranosyl residue in unit 1 and O-factor III seems to be due to the same groups. The chemical natures of the determinants responsible for O-factors 4 and 3, 4 are still obscure. The structural studies indicate that the immunological classification of Sh. flexneri serotypes and subserotypes, as regards these O-factors, may need revision.

https://febs.onlinelibrary.wiley.com/doi/pdf/10.1111/j.1432-1033.1978.tb20963.x

Structural studies of Shigella flexneri O-antigens.

Structural studies of the O-specific side-chains of the shigella sonnei phase I lipopolysaccharide

The rate of coupling of oligosaccharides having aldose end groups to protein by reductive amination was significantly increased by changing the temperature and pH of the reaction, and even more sig...

Improved procedures for the conjugation of oligosaccharides to protein by reductive amination

Capsular polysaccharides of types Ia, Ib, II and IIIGroup B Streptococcus and groups B and C Neisseria meni ngi ti di S contai n termi nal si al i c aci d i n di ffe rent molecular environments. Experimentation has identified sialic acid as an important factor in the virulence of these organisms and in the human antibody response to their capsular polysacchari de anti gens . Al though termi nal si al i c aci d i s not normally immunogenic it controls the determinants which are responsible for the production of protective antibodies. Using immunological and NMR spectroscopic techniques on the native and specifically modified polysaccharides, a number of these si al I c aci d-control 1 ed determi nants have been identified and located. These determinants are only formed in structures which can accommodate long—range interactions between sialic acid and other remote glycosyl residues. The carboxylate group of si ali c acid is essential for these interactions to occur.

Structure, conformation and immunology of sialic acid-containing polysaccharides of human pathogenic bacteria

The review presents the structures of the O-specific polysaccharides (O-antigens) of the lipopolysaccharides isolated from over 25 Citrobacter strains, which represent different species and serogroups. The correlation between O-antigen structure and immunospecificity as well as numerous cross-reactions between Citrobacter and other enterobacterial species are discussed.

/pdf/structures-and-serology-of-the-o-specific-polysaccharides-of-34z8atado9.pdf

Ewa Katzenellenbogen

Papers

Structural studies of Shigella flexneri O-antigens.

Structural studies of the O-specific side-chains of the shigella sonnei phase I lipopolysaccharide

Improved procedures for the conjugation of oligosaccharides to protein by reductive amination

Structure, conformation and immunology of sialic acid-containing polysaccharides of human pathogenic bacteria

Structures and serology of the O-specific polysaccharides of bacteria of the genus Citrobacter.