Glycosphingolipids with Lewis Blood Group Activity: Uptake by Human Erythrocytes

Marcus, Donald M.; Cass, Louise E.

doi:10.1126/science.164.3879.553

Cited by 264 publications

(89 citation statements)

References 24 publications

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“…LeX and sLeX are members of the Lewis system histo-blood group antigens, which were originally identified on RBCs, where they are acquired from the plasma as glycolipids (22). A common method for assessing Ply activity is by determining the toxin's ability to lyse RBCs in hemolytic assays.…”

Section: Resultsmentioning

confidence: 99%

“…The Lewis antigens were originally identified on human erythrocytes, in plasma, and in mucous secretions, where they were assumed to be glycoproteins. It was later shown that RBCs passively acquire Lewis antigens from plasma, predominantly as glycosphingolipids, and incorporate them into the erythrocyte membrane (22). The type 1 Lewis antigens, LeA and LeB, are important histo-blood group antigens, whereas the type 2 Lewis antigens, LeX and LeY, are expressed at relatively low levels in normal tissues but are found to be overexpressed on the surface of human tumor cells from various sites (27).…”

Section: Discussionmentioning

confidence: 99%

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The cholesterol-dependent cytolysins pneumolysin and streptolysin O require binding to red blood cell glycans for hemolytic activity

Shewell

Harvey

Higgins

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

105

113

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The cholesterol-dependent cytolysin (CDC) pneumolysin (Ply) is a key virulence factor of Streptococcus pneumoniae. Membrane cholesterol is required for the cytolytic activity of this toxin, but it is not clear whether cholesterol is the only cellular receptor. Analysis of Ply binding to a glycan microarray revealed that Ply has lectin activity and binds glycans, including the Lewis histoblood group antigens. Surface plasmon resonance analysis showed that Ply has the highest affinity for the sialyl LewisX (sLeX) structure, with a K d of 1.88 × 10 −5 M. Ply hemolytic activity against human RBCs showed dose-dependent inhibition by sLeX. Flow cytometric analysis and Western blots showed that blocking binding of Ply to the sLeX glycolipid on RBCs prevents deposition of the toxin in the membrane. The lectin domain responsible for sLeX binding is in domain 4 of Ply, which contains candidate carbohydrate-binding sites. Mutagenesis of these predicted carbohydrate-binding residues of Ply resulted in a decrease in hemolytic activity and a reduced affinity for sLeX. This study reveals that this archetypal CDC requires interaction with the sLeX glycolipid cellular receptor as an essential step before membrane insertion. A similar analysis conducted on streptolysin O from Streptococcus pyogenes revealed that this CDC also has glycan-binding properties and that hemolytic activity against RBCs can be blocked with the glycan lacto-N-neotetraose by inhibiting binding to the cell surface. Together, these data support the emerging paradigm shift that pore-forming toxins, including CDCs, have cellular receptors other than cholesterol that define target cell tropism.S treptococcus pneumoniae is a leading cause of morbidity and mortality worldwide. This bacterial pathogen is responsible for a range of diseases, including pneumonia, meningitis, septicemia, and otitis media. One of the major virulence factors of S. pneumoniae is the multifunctional pore-forming toxin pneumolysin (Ply). Ply is produced by virtually all clinical isolates of S. pneumoniae and is a member of the cholesterol-dependent cytolysin (CDC) family of toxins (1). The key feature of the CDCs, which are expressed by a number of pathogenic Grampositive bacteria, is the ability to form pores in cholesterolcontaining cell membranes. The pore-forming mechanism of the CDCs is a multistep process that involves recognition and binding to the cholesterol-containing membrane by domain 4 of the toxin, oligomerization of ∼34-50 soluble monomers on the target cell membrane to form a large prepore complex (2), and penetration of the prepore structure into the membrane to become a transmembrane β-barrel pore (3-5).The cytolytic mechanism of the CDCs depends on the presence of cholesterol in the target cell membrane; hence, it was thought that cholesterol served as the cellular receptor for these toxins. The first suggestion of this cholesterol serving as the receptor occurred in the 1970s, when it was found that preincubation of the CDC of Streptococcus pyogenes, streptolysin O (SL...

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

The cholesterol-dependent cytolysins pneumolysin and streptolysin O require binding to red blood cell glycans for hemolytic activity

Shewell

Harvey

Higgins

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

105

113

View full text Add to dashboard Cite

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“…Compared to other erythrocyte blood-group systems the Lewis blood group system is unique as the Lewis active glycolipids are not synthesised in the erythrocytes but acquired from plasma (Sneath & Sneath, 1959;Marr et al, 1967;Marcus & Cass, 1969). The Lea antigen determinant GlcNAc-R) is synthesised by the action of an al-4-L-fucosyltransferase encoded by the Lewis gene (Shen, Grollman & Ginsburg, 1968), and the activity of this enzyme can be determined in saliva.…”

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confidence: 99%

Frequency and mechanism of Lewis antigen expression in human urinary bladder and colon carcinoma patients

Langkilde¹,

Wolf²,

Meldgård³

et al. 1991

Br J Cancer

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Changes in the expression of Lewis antigens have been associated with cancer diseases, and recent results have pointed at a possible increased risk of cancer development among Lewis negative patients. The frequency of the erythrocyte Lewis phenotypes Le(a- b+), Le(a+ b-) and Le(a- b-) was analysed in patients suffering from urinary bladder cancer (82), colon cancer (21), and benign urological diseases (45). An increased frequency of Lewis negative individuals was found among colon cancer patients (P less than 0.004) and bladder cancer patients (P = 0.05). The Lewis negative phenotype was shown to be associated with unfavourable disease parameters: invasion (P less than 0.02) and high grade of atypia (P less than 0.01) in bladder cancer patients, and high Dukes stage (P less than 0.05) in colon cancer patients. alpha 1-4fucosyltransferase activity (Lewis transferase) was shown to be present in saliva from four out of eight erythrocyte Lewis negative cancer patients, indicating that some patients with advanced cancer disease may have converted from a Lewis positive to a Lewis negative phenotype.

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“…Le a1 and Le b (2, 3) are the major Lewis antigens found on human erythrocytes. These fucosylated glycosphingolipids are synthesized by exocrine epithelial cells (4) and are secondarily passively adsorbed onto erythrocytes in the peripheral circulation giving these blood cells their Lewis phenotype (5).…”

mentioning

confidence: 99%

Significance of Individual Point Mutations, T202C and C314T, in the Human Lewis (FUT3) Gene for Expression of Lewis Antigens by the Human α(1,3/1,4)-Fucosyltransferase, Fuc-TIII

Elmgren¹,

Mollicone²,

Costache³

et al. 1997

Journal of Biological Chemistry

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The Lewis ␣(1,3/1,4)-fucosyltransferase, Fuc-TIII, encoded by the FUT3 gene is responsible for the final synthesis of Le a and Le b antigens. Various point mutations have been described explaining the Lewis negative phenotype, Le(a؊b؊), on erythrocytes and secretions. Two of these, T202C and C314T originally described in a Swedish population, have not been found as single isolated point mutations so far. To define the relative contribution of each of these two mutations to the Lewis negative phenotype, we cloned and made chimeric FUT3 constructs separating the T202C mutation responsible for the amino acid change Trp 68 3 Arg, from the C314T mutation leading to the Thr 105 3 Met shift. COS-7 cells were transfected and the expression of Fuc-TIII enzyme activity and the presence of Lewis antigens were determined. There was no decrease in enzyme activity nor of immunofluorescence staining on cells transfected with the construct containing the isolated C314T mutation compared with cells transfected with a wild type FUT3 allele control. No enzyme activity nor immunoreactivity for Lewis antigens was detected in FUT3 constructs containing both mutations in combination. The T202C mutation alone decreased the enzyme activity to less than 1% of the activity of the wild type FUT3 allele. These results demonstrate, that the Trp 68 3 Arg substitution in human Fuc-TIII is the capital amino acid change responsible for the appearance of the Le(a؊b؊) phenotype on human erythrocytes in individuals homozygous for both the T202C and C314T mutations.The Lewis histo-blood group system comprises different complex carbohydrate structures, which participate in different biological processes such as embryogenesis, tissue differentiation, tumor metastasis, inflammation, and bacterial adhesion (1). Le a1 and Le b (2, 3) are the major Lewis antigens found on human erythrocytes. These fucosylated glycosphingolipids are synthesized by exocrine epithelial cells (4) and are secondarily passively adsorbed onto erythrocytes in the peripheral circulation giving these blood cells their Lewis phenotype (5).It was shown as early as the 1950's, that the Lewis phenotype of erythrocytes is influenced by the ABH secretor status of the individual (6), and the erythrocyte phenotype is the result of the epistatic interaction of the Lewis (Le-le) and the salivary ABH secretor (Se-se) loci (7). Fucosyltransferases (Fuc-Ts) encoded by genes at these loci compete and interact with each other and with other glycosyltransferases to determine the individual's final Lewis and secretor phenotypes.Since 1990, seven human fucosyltransferase genes (FUT1-7) have been cloned, sequenced, and characterized according to acceptor specificities (8 -18). The human ␣(1,2)-Fuc-T gene family comprises FUT1 encoding for the H enzyme and FUT2 encoding for the human secretor ␣(1,2)-fucosyltransferase (9). FUT3-7 encode for ␣(1,3)-Fuc-Ts. Two of these human enzymes, respectively (10,14), also express ␣(1,4)-fucosyltransferase activities (15,19,20).Five main missense mutations have be...

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Glycosphingolipids with Lewis Blood Group Activity: Uptake by Human Erythrocytes

Cited by 264 publications

References 24 publications

The cholesterol-dependent cytolysins pneumolysin and streptolysin O require binding to red blood cell glycans for hemolytic activity

The cholesterol-dependent cytolysins pneumolysin and streptolysin O require binding to red blood cell glycans for hemolytic activity

Frequency and mechanism of Lewis antigen expression in human urinary bladder and colon carcinoma patients

Significance of Individual Point Mutations, T202C and C314T, in the Human Lewis (FUT3) Gene for Expression of Lewis Antigens by the Human α(1,3/1,4)-Fucosyltransferase, Fuc-TIII

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