Fuc(α1→3)GalNAc-: the major antigenic motif of Schistosoma mansoni glycolipids implicated in infection sera and keyhole-limpet haemocyanin cross-reactivity

Kantelhardt, Sven R.; Wuhrer, Manfred; Dennis, Roger D.; Doenhoff, Michael J.; Bickle, Q. D.; Geyer, Rudolf

doi:10.1042/bj20011678

Cited by 52 publications

(60 citation statements)

References 27 publications

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“…1). In contrast to rabbit hyperimmune serum raised against KLH, antibody binding was mostly eliminated by mild HF treatment, which results in a release of fucose residues leaving the remaining carbohydrate chains largely intact (23). In agreement with previous data (22) it may be, therefore, concluded that cross-reacting oligosaccharides comprise a fucosylated carbohydrate epitope.…”

Section: Isolation Of Klh Oligosaccharides Sharing Carbohydrate Epitosupporting

confidence: 78%

Identification and Characterization of Keyhole Limpet Hemocyanin N-Glycans Mediating Cross-reactivity with Schistosoma mansoni

Geyer

Wuhrer

Resemann

et al. 2005

Journal of Biological Chemistry

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Keyhole limpet hemocyanin (KLH) of the mollusc Megathura crenulata is known to serologically cross-react with Schistosoma mansoni glycoconjugates in a carbohydrate-dependent manner. To elucidate the structural basis for this cross-reactivity, KLH glycans were released from tryptic glycopeptides and fluorescently labeled. Cross-reacting glycans were identified using a polyclonal antiserum reacting with soluble S. mansoni egg antigens, isolated by a threedimensional fractionation scheme and analyzed by different mass spectrometric techniques as well as linkage analysis and exoglycosidase treatment. The results revealed that cross-reacting species comprise ϳ4.5% of released glycans. They all represent novel types of N-glycans with a Fuc(␣1-3)GalNAc(␤1-4)[Fuc(␣1-3)]GlcNAc motif, which is known to occur also in schistosomal glycoconjugates. The tetrasaccharide unit is attached to the 3-linked antenna of a trimannosyl core, which can be further decorated by galactosyl residues, a xylose residue in 2-position of the central mannose and/or a fucose at the innermost N-acetylglucosamine. This study provides for the first time detailed structural data on the KLH carbohydrate entities responsible for cross-reactivity with glycoconjugates from S. mansoni.Hemocyanins act as oxygen-transporting proteins in many arthropod and mollusc species (1). In addition to this biochemical function, the hemocyanin of the Californian giant keyhole limpet Megathura crenulata, a marine gastropod, is known to be a potent immunoactivator (for reviews, see Refs. 2 and 3). Due to these immunostimulatory properties, keyhole limpet hemocyanin (KLH) 2 is widely used in research and clinical studies. Present fields of application are, for example, immunotherapy of bladder cancer (4 -7), based on the assumed expression of Gal(␤1-3)GalNAc determinants as cross-reacting epitopes (8), and its use as a carrier of polysialic acid or low molecular weight haptens, such as synthetic oligosaccharides, gangliosides, or (glyco)peptides, designed for potential application in anticancer therapy (9 -14). Intriguingly, KLH has been further demonstrated to exhibit a carbohydrate-based cross-reactivity with glycoconjugates from Schistosoma mansoni (15) thus allowing the diagnosis of S. mansoni (6, 7, 16), Schistosoma hematobium (17), and Schistosoma japonicum (18) infections by enzymelinked immunosorbent assay (ELISA). Hence, KLH has also been discussed as a potential candidate for vaccination against schistosomiasis (15). On the other hand, agarose beads coated with KLH or KLH-glycopeptides have been shown to mimic in a carbohydrate-dependent manner the hepatic granuloma formation mediated by S. mansoni eggs (19). Therefore, KLH may also be regarded as a model antigen to study the immunopathological mechanisms of schistosome infections.Although it is generally accepted that the oligosaccharide constituents of KLH are of prime significance for its antigenicity and biomedical properties (2), knowledge on the carbohydrate structure of this glycoprotein is still incomp...

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Section: Isolation Of Klh Oligosaccharides Sharing Carbohydrate Epitosupporting

confidence: 78%

Identification and Characterization of Keyhole Limpet Hemocyanin N-Glycans Mediating Cross-reactivity with Schistosoma mansoni

Geyer

Wuhrer

Resemann

et al. 2005

Journal of Biological Chemistry

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“…Given the robust expression via immunostaining in schistosomula and adults (Fig. 6B), it is possible that the glycan epitope also is expressed on glycolipids, which is consistent with previous reports of lipid antigens (13,(63)(64)(65)(66).…”

Section: Preparation Of N-glycans From Schistosoma Mansoni Eggssupporting

confidence: 78%

Identification of Antigenic Glycans from Schistosoma mansoni by Using a Shotgun Egg Glycan Microarray

et al. 2016

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f Infection of mammals by the parasitic helminth Schistosoma mansoni induces antibodies to glycan antigens in worms and eggs, but the differential nature of the immune response among infected mammals is poorly understood. To better define these responses, we used a shotgun glycomics approach in which N-glycans from schistosome egg glycoproteins were prepared, derivatized, separated, and used to generate an egg shotgun glycan microarray. This array was interrogated with sera from infected mice, rhesus monkeys, and humans and with glycan-binding proteins and antibodies to gather information about the structures of antigenic glycans, which also were analyzed by mass spectrometry. A major glycan antigen targeted by IgG from different infected species is the FLDNF epitope [Fuc␣3GalNAc␤4(Fuc␣3)GlcNAc-R], which is also recognized by the IgG monoclonal antibody F2D2. The FLDNF antigen is expressed by all life stages of the parasite in mammalian hosts, and F2D2 can kill schistosomula in vitro in a complement-dependent manner. Different antisera also recognized other glycan determinants, including core ␤-xylose and highly fucosylated glycans. Thus, the natural shotgun glycan microarray of schistosome eggs is useful in identifying antigenic glycans and in developing new anti-glycan reagents that may have diagnostic applications and contribute to developing new vaccines against schistosomiasis. Schistosomiasis is a major health problem in tropical and subtropical areas where it is endemic, with more than 200 million people actively infected and 800 million at risk of contracting the disease (1-3). Current treatment for disease is limited to the drug praziquantel (4), but cases of drug resistance have been reported (5). Decades of research on schistosomiasis vaccines have yielded only two candidates for clinical trials, and no encouraging results have been published yet (6-9). Thus, there is an urgent need to develop more sensitive diagnostic methods and to identify new vaccine candidates.Recent studies have shown that a major part of the host immune response to infection is directed against carbohydrate (glycan) antigens in glycoproteins and glycolipids (10-17). A wide variety of unusual antigenic determinants include glycans containing the LDN, fucosylated LDN sequences (LDNF, LDN-dF, FLDN, and FLDNF), Lewis X (Le x ), poly-Le x , core ␣3 fucose, and core ␤2 xylose structures (Fig. 1) (11, 14, 17), many of which are expressed by all developmental stages of schistosomes (18). Interestingly, monoclonal antibodies (MAbs) specific to these glycans recognize these antigens on the surface of 3-h-old schistosomula, and some anti-glycan antibodies can mediate killing in vitro in a complement-dependent fashion (18-21). Schistosoma mansoniinfected rhesus monkeys, which are known to self-cure after infection, have IgG to many glycan antigens, including Le x , LDN, LDNF, core fucose, and core xylose determinants, and their sera are effective in complement-mediated cytolysis of cells expressing Le x as well as schistosomulum larvae in ...

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“…Although the nature of the oligosaccharides on the surface of KLH has not been completely elucidated, those identified to date have been shown to contain unique oligosaccharide patterns, resulting in novel structures (45,46). These novel structures, together with the high levels of cross-reactivity with a variety of carbohydrate Ags, may contribute to the immunogenicity of KLH (47).…”

Section: Discussionmentioning

confidence: 99%

“…The cross-reactive epitope was identified (49), and clinical trials with Immucothel, a KLH subunit, are reported (50). In addition, a fucose epitope on the surface of S. mansoni has been found to be cross-reactive with KLH (46,51). Immobilized KLH or anti-KLH Abs have been used to diagnose acute vs chronic infections of S. mansoni (52).…”

Section: Discussionmentioning

confidence: 99%

Antibodies to Keyhole Limpet Hemocyanin Cross-React with an Epitope on the Polysaccharide Capsule of Cryptococcus neoformans and Other Carbohydrates: Implications for Vaccine Development

May

Beenhouwer

Scharff

2003

The Journal of Immunology

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Cryptococcus neoformans causes a life-threatening meningoencephalitis in AIDS patients. Mice immunized with a glycoconjugate vaccine composed of the glucuronoxylomannan (GXM) component of the cryptococcal capsular polysaccharide conjugated to tetanus toxoid produce Abs that can be either protective or nonprotective. Because nonprotective Abs block the efficacy of protective Abs, an effective vaccine must focus the Ab response on a protective epitope. Mice immunized with peptide mimetics of GXM conjugated to keyhole limpet hemocyanin (KLH) with glutaraldehyde developed Abs to GXM. However, control peptides P315 and P24 conjugated to KLH also elicited Abs to GXM. GXM-binding Abs from mice immunized with P315-KLH were inhibited by KLH treated with glutaraldehyde (KLH-g), but not by P315. Furthermore, KLH-g inhibited binding of GXM by serum of mice immunized with GXM-TT, indicating that glutaraldehyde treatment of KLH reveals an epitope(s) that cross-reacts with GXM. Vaccination with KLH-g or unmodified KLH elicited Abs to GXM, but did not confer protection against C. neoformans, suggesting the cross-reactive epitope on KLH was not protective. This was supported by the finding that 4H3, a nonprotective mAb, cross-reacted strongly with KLH-g. Sera from mice immunized with either native KLH or KLH-g cross-reacted with several other carbohydrate Ags, many of which have been conjugated to KLH for vaccine development. This study illustrates how mAbs can be used to determine the efficacy of potential vaccines, in addition to describing the complexity of using KLH and glutaraldehyde in the development of vaccines to carbohydrate Ags.

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Fuc(α1→3)GalNAc-: the major antigenic motif of Schistosoma mansoni glycolipids implicated in infection sera and keyhole-limpet haemocyanin cross-reactivity

Cited by 52 publications

References 27 publications

Identification and Characterization of Keyhole Limpet Hemocyanin N-Glycans Mediating Cross-reactivity with Schistosoma mansoni

Identification and Characterization of Keyhole Limpet Hemocyanin N-Glycans Mediating Cross-reactivity with Schistosoma mansoni

Identification of Antigenic Glycans from Schistosoma mansoni by Using a Shotgun Egg Glycan Microarray

Antibodies to Keyhole Limpet Hemocyanin Cross-React with an Epitope on the Polysaccharide Capsule of Cryptococcus neoformans and Other Carbohydrates: Implications for Vaccine Development

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