Neuere Ergebnisse zur Biochemie der Zellwand‐Lipopolysaccharide von Salmonella‐Bakterien

Lüderitz, Otto

doi:10.1002/ange.19700821802

Cited by 16 publications

(7 citation statements)

References 54 publications

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“…These antibodies are mainly of GIcNAc specificity and are restricted predominantly to the IgG2/K isotype (7). Anti-GlcNAc antibodies may have been induced not only by A-CHO, but also by other bacterial polysaccharides containing terminal GIcNAc moieties in a 1,3-glycosidic linkage, such as the O substance of gram-negative bacteria (30). Moreover, terminal GIcNAc is exposed transiently during synthesis or degradation of blood group substance (P. Hanfland, personal communication) and might act as an antigenic stimulus.…”

Section: To Demonstrate That the Increase Of Anti-a-cho Antibodies Wamentioning

confidence: 99%

Human immune response to group A streptococcal carbohydrate (A-CHO). I. Quantitative and qualitative analysis of the A-CHO-specific B cell population responding in vitro to polyclonal and specific activation.

Emmrich¹,

Schilling²,

Eichmann³

1985

The Journal of Experimental Medicine

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The immune response to the group-specific carbohydrate of group A streptococci (A-CHO) provides an informative in vitro model for the investigation of several aspects of human anticarbohydrate immune responses. A-CHO-specific B cells can be polyclonally activated by pokeweed mitogen (PWM), and, specifically, by in vitro immunization with streptococcal vaccine. High levels of A-CHO-specific antibodies, mainly directed to the immunodominant side chain N-acetyl-D-glucosamine (GlcNAc), occur in healthy adult individuals. Serum antibody levels are reflected in high frequencies of precursor B cells among peripheral blood lymphocytes. In one particular case, greater than 15% of all B cells activated by PWM for IgM production were found to produce IgM anti-A-CHO antibodies, as determined in limiting dilution experiments, as well as by analyzing Ig concentrations in bulk culture experiments. The case with the lowest proportion observed had 0.3% A-CHO-specific B cells among IgM-producing B cells. Preferential PWM activation of anti-A-CHO-producing B cells could be excluded. The comparison of the proportions of anti-A-CHO IgM produced in vivo, and of B cells producing antibodies of this specificity in peripheral blood, suggests a similar distribution of specific precursor B cells in the antibody-producing lymphoid tissue compartments and in peripheral blood. However, nearly all specific antibodies produced in vitro belong to the IgM isotype, whereas IgG anti-A-CHO in high amounts, mostly exceeding the specific IgM, was found only among anti-A-CHO antibodies produced in vivo. Low anti-A-CHO IgG production was seen in polyclonally activated as well as in antigen-activated cultures, whereas, in contrast, total IgG was produced in considerable amounts after polyclonal activation. This suggests a different distribution pattern, and/or diverse differentiation requirements for anti-A-CHO-producing B cells, compared with other B cell species.

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Section: To Demonstrate That the Increase Of Anti-a-cho Antibodies Wamentioning

confidence: 99%

Human immune response to group A streptococcal carbohydrate (A-CHO). I. Quantitative and qualitative analysis of the A-CHO-specific B cell population responding in vitro to polyclonal and specific activation.

Emmrich¹,

Schilling²,

Eichmann³

1985

The Journal of Experimental Medicine

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“…Fraction P+ is, therefore, present as such in the lipopolysaccharide and does not originate as a degradation product of fraction PPN. DISCUSSION Although much has been learned in the past decade about the structure of O-specific chains and the hexose region of the core of Salmonella cell wall lipopolysaccharides [26], there is a great lack of knowledge concerning the inner part of the core. This region, which contains heptose and KDO, and which connects the polysaccharide chain with lipid A, has been investigated only in a special class of R mutants, the so-called P-mutants [3,27].…”

Section: Identification Of Fraction Ppnmentioning

confidence: 99%

The Linkage of Pyrophosphorylethanolamine to Heptose in the Core of Salmonella minnesota Lipopolysaccharides

Lehmann¹,

Lüderitz²,

Westphal³

1971

European Journal of Biochemistry

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Mild acid hydrolysis of the P+ lipopolysaccharide ol' Salmonella minnesota mRz, a UDPglucose synthetase-less mutant, leads to the formation of 5 degradation products whose structures were studied. Free 2-keto-3-deoxyoctonate (KDO), KDO-7-phosphorylethanolamine, and fraction P-(Hep-Hep-KDO) have been identified previously as degradation products of a Plipopolysaccharide (glycolipid). Fractions P+ (Hep-(Hep-4-phosphate)-KDO) and P P N (Hep-(Hep-4-pyrophosphorylethanolamine)-KDO) occur only i i i the P+ mutant. It was shown that fraction Pf is not an artifact which was formed from fraction PPN during partial hydrolysis. The simultaneous occurrence of fractions P+ and PPN reflects microheterogenicity of the lipopolysaccharide. Possible reasons for this are discussed.Salmonella R forms synthesize defective lipopolysaccharides consisting of lipid A to which are linked smaller or larger parts of the core polysaccharide [I]. Two classes of mutants, designated as P-and P f , have been identified [I, 21. Lipopolysaccharides of Pmutants, which were designated as glycolipids [Z], can be recognized by their relatively lower phosphate content. On acid hydrolysis free heptose and ethanolamine (and only small amounts of phosphorylethanolamine) are liberated. It has been shown that this ethanolamine derives a t least partly from the core where it is present as a phosphorylethanolamine group linked to the 7-position of a 2-keto-3-deoxyoctonate residue [3].I n contrast, lipopolysaccharides of P+ mutants contain relatively more phosphate, and on hydrolysis heptosephosphate and phosphorylethanolamine (besides ethanolamine) are liberated.The present paper describes investigations performed with lipopolysaccharides of Salmonella Pf mutants. The results show that the excess of phosphate is present as phosphate or pyrophosphorylethanolamine groups which are linked to the 4 position of heptose I units of the core. MATERIAL AND METHODS Bacteria and LipopolysaccharidesThe Salmonella minnesota R mutant mRz is a UDP-glucose synthetase-less mutant of the chemotype Rd,P+ [4]. The S. minnesota R mutant mR7 belongs to the chemotype RdL,P-due to defects in the transfer of glucose I and of phosphate groups to heptose units [2,4].Unusual Abbreviation. KDO, 2-keto-3-deoxyoctonate.P+ lipopolysaccharides and P-glycolipids were isolated by the phenol-chloroform-petroleum ether and purified by ultracentrifugation [5,6]. Analytical MethodsFor :full details of these methods see [3]. Heptose and maiinose were determined with H,SO,/cysteine [7], in s'ome cases mannose was also estimated with mannose isomerase according to Palleroni and Doudoroff [El]. Glucose was determined with glucose oxidase, 2-keto-3-deoxy-octonate (KDO) with periodate/tliiobarbituric acid [9] N NaOH [17]. After I0 min a t room temperature 100 pl of 0.1 N NaOH were added and the mixture allowed to stand at; room temperature for a further 30min. The solution was adjusted to pH 4 with acetic acid and extracted twice with chloroform. The water phase wi%s evaporated to dryness and salt re...

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“…The S. minnesota mutants mR5 (RcP-) and mR7 (RdP-) were isolated by J. Schlosshardt (Zentrallaboratorium fur Bakterielle Darminfektionen, Berlin-Pankow, DDR). For definition of chemotypes, Rc, Rd, etc., see [1,15]. The core structures and defects of these mutants are listed in Table 1.…”

Section: Organismsmentioning

confidence: 99%

Biosynthesis of Salmonella Lipopolysaccharide

Mühlradt¹

1971

European Journal of Biochemistry

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Several transfer reactions involved in the biosynthesis of the core part of Salmonella lipopolysaccharide have been studied in a cell free system. This system contained three components: (1) enzymes which were solubilised by repeated washings of EDTA‐lysozyme spheroplasts; (2) acceptor which consisted of either a heated cell wall fraction sedimenting at 20 000 x g, or heated, partially lysed, EDTA‐lysozyme treated cells; and (3) radioactively labelled precursors ([32P]ATP, UDP‐[14C]Glc, UDP‐[14C]Gal). The following results were obtained with this transfer system: (1) only the γ‐phosphate group of ATP is transferred to the heptose moiety of the core; (2) polyphosphate which is synthesised enzymatically in a a side‐reaction by the system is not involved in the heptose phosphorylation; (3) there is a definite sequence of addition of the core substituents glucose, phosphate, and galactose to the incomplete core stub. This sequence is as follows: (Hep)2‐(KDO, ethanolamine, phosphate, lipid A) → Glc‐(Hep)2‐(KDO, ethanolamine, phosphate, lipid A) → Glc‐(Hep)2P‐(KDO, ethanolamine, phosphate, lipid A) → (Gal)2‐Glc‐(Hep)2P‐(KDO, ethanolamine, phosphate, lipid A).

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Neuere Ergebnisse zur Biochemie der Zellwand‐Lipopolysaccharide von Salmonella‐Bakterien

Cited by 16 publications

References 54 publications

Human immune response to group A streptococcal carbohydrate (A-CHO). I. Quantitative and qualitative analysis of the A-CHO-specific B cell population responding in vitro to polyclonal and specific activation.

Human immune response to group A streptococcal carbohydrate (A-CHO). I. Quantitative and qualitative analysis of the A-CHO-specific B cell population responding in vitro to polyclonal and specific activation.

The Linkage of Pyrophosphorylethanolamine to Heptose in the Core of Salmonella minnesota Lipopolysaccharides

Biosynthesis of Salmonella Lipopolysaccharide

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