A new monoclonal antibody (A46-B/B10) highly specific for the blood group H type 2 epitope: Generation, epitope analysis, serological and histological evaluation

Karsten, Uwe; Pilgrim, G; Hanisch, Franz‐Georg; Uhlenbruck, G.; Kasper, Maren; Stosiek, P; Papsdorf, G; Pasternak, G

doi:10.1038/bjc.1988.187

Cited by 13 publications

(9 citation statements)

References 23 publications

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“…2-dGal also interferes with the maintenance of long-term potentiation (LTP), a form of synaptic plasticity that is closely associated with learning and memory (4). Moreover, injection of a monoclonal antibody specific for Fuc␣(1-2)Gal has been found to impair memory formation in animals, presumably by blocking the Fuc␣(1-2)Gal epitope (5,6). These studies suggest important roles for Fuc␣(1-2)Gal carbohydrates and their associated proteins in modulating neuronal communication in the brain.…”

mentioning

confidence: 99%

Protein fucosylation regulates synapsin Ia/Ib expression and neuronal morphology in primary hippocampal neurons

Murrey

Gama

Kalovidouris

et al. 2005

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

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Although fucose-␣(1-2)-galactose [Fuc␣(1-2)Gal] carbohydrates have been implicated in cognitive processes such as long-term memory, the molecular mechanisms by which these sugars influence neuronal communication are not well understood. Here, we present molecular insights into the functions of Fuc␣(1-2)Gal sugars, demonstrating that they play a role in the regulation of synaptic proteins and neuronal morphology. We show that synapsins Ia and Ib, synapse-specific proteins involved in neurotransmitter release and synaptogenesis, are the major Fuc␣(1-2)Gal glycoproteins in mature cultured neurons and the adult rat hippocampus. Fucosylation has profound effects on the expression and turnover of synapsin in cells and protects synapsin from degradation by the calcium-activated protease calpain. Our studies suggest that defucosylation of synapsin has critical consequences for neuronal growth and morphology, leading to stunted neurite outgrowth and delayed synapse formation. We also demonstrate that Fuc␣(1-2)Gal carbohydrates are not limited to synapsin but are found on additional glycoproteins involved in modulating neuronal architecture. Together, our studies identify important roles for Fuc␣(1-2)Gal sugars in the regulation of neuronal proteins and morphological changes that may underlie synaptic plasticity., which exists as a terminal carbohydrate modification to N-and O-linked glycoproteins, has been implicated in cognitive processes such as learning and memory. For instance, preventing formation of Fuc␣(1-2)Gal linkages by incorporation of 2-deoxy-D-galactose (2-dGal) into glycan chains has been shown to cause reversible amnesia in animals (1-3). 2-dGal also interferes with the maintenance of long-term potentiation (LTP), a form of synaptic plasticity that is closely associated with learning and memory (4). Moreover, injection of a monoclonal antibody specific for Fuc␣(1-2)Gal has been found to impair memory formation in animals, presumably by blocking the Fuc␣(1-2)Gal epitope (5, 6). These studies suggest important roles for Fuc␣(1-2)Gal carbohydrates and their associated proteins in modulating neuronal communication in the brain.Interestingly, evidence suggests that protein fucosylation is regulated in response to synaptic activity. Both task-specific learning and LTP have been shown to induce the fucosylation of proteins at the synapse (7,8), and addition of exogenous fucose or 2Јfuco-syllactose was found to enhance LTP in hippocampal slices (9). The activity of fucosyltransferases, enzymes involved in the transfer of fucose to glycoproteins, has also been demonstrated to increase substantially during synaptogenesis (10) and upon passive avoidance training in animals (11). Together, these studies suggest that protein fucosylation may be a highly regulated process in the brain and may contribute to neuronal development and synaptic plasticity. Despite these intriguing observations, little is known about the molecular mechanisms by which Fuc␣(1-2)Gal sugars influence neuronal communication. Surprisingly, no Fuc␣(...

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confidence: 99%

Protein fucosylation regulates synapsin Ia/Ib expression and neuronal morphology in primary hippocampal neurons

Murrey

Gama

Kalovidouris

et al. 2005

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

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“…Therefore, it is not surprising that some of them are also tumour markers, or more precisely oncofetal antigens [37]. The Lewis antigens, and in particular Lewis Y, Lewis X and some H‐related structures belong to this category [30, 31, 38]. Among these antigens, Lewis Y is of special interest as a tumour‐associated antigen with potential applications in diagnosis and therapy.…”

Section: Discussionmentioning

confidence: 99%

“…Although there were some variations in the amino acids influencing the canonical structures of the CDRs, they were not exceeding the allowed constrained variation for any of the CDRs, leaving the binding specificity only affected by the side chain type [35]. The antibodies MSL5 [19] and A46-B ⁄ B10 [30] are almost completely identical and constitute a third sequence set.…”

Section: Discussionmentioning

confidence: 99%

“…Antibody production. A70‐A/A9 (IgG1, κ) recognizing Lewis Y and partially Lewis b [29], A46‐B/B10 (IgM, κ) recognizing Lewis Y and H type 2 [30, 31] and A70‐C/C8 (IgM, κ) recognizing Lewis Y [31] have been published. A51‐B/A6 (IgA, κ) recognizing Lewis Y, H disaccharide and H type 2 was generated using a sequence of different glycoconjugates and MCF‐7 cells as immunogens.…”

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Modifying Antibody Specificity by Chain Shuffling of V_H / V_L between Antibodies with Related Specificities

et al. 2008

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Histo‐blood group antigens are important markers of developmental stages and as such also often of tumours. Generation of antibodies towards these carbohydrate structures is still a challenging task as they may lack specificity, affinity or are only of the IgM class. We have examined four own antibodies to Lewis Y/H type 2 for their fine specificities using a large panel of mono‐ and oligosaccharides. Sequence alignment to other antibodies with similar specificity revealed an overall limited variation, and that our antibodies constitute a novel set. Based on produced and analysed chimeric mouse–human antibodies, extensive chain shuffling experiments were performed in order to analyse influences of the respective H and L chains on the specificity of the antibodies, and to generate modified antibodies with improved properties. One chIgG1 out of the shuffled antibodies revealed improved specificity and markedly enhanced functional affinity to Lewis Y compared to the parental chIgG1 antibodies. Therefore, the combinatorial approach of chain shuffling provides a platform to improve specificity and/or affinity of anti‐carbohydrate antibodies.

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“…Wrapped slides could be stored at −80°C until use. For immunocytochemistry, cells were fixed with cold (−20°C) acetone for 15 min, blocked with 2% bovine serum albumin (BSA) for 30 min, and incubated with CD44 mAb (G44-26, mouse IgG2b, BD Biosciences, Franklin Lakes, NJ, USA) together with mAbs CD173 (A46-B/B10, mouse IgM) [11] or CD174 (A70-C/C8, mouse IgM), both from Glycotope GmbH (Berlin, Germany) for 60 min. The slides were subsequently incubated with a mixture of fluorescein isothiocyanate (FITC)-conjugated anti-mouse IgM (μ chain-specific; F9259, Sigma) and Cy3-conjugated goat anti-mouse IgG (γ chain-specific; #69732, Jackson Laboratories, West Grove, PA, USA).…”

Section: Immunocytochemistrymentioning

confidence: 99%

Co-expression of CD173 (H2) and CD174 (Lewis Y) with CD44 suggests that fucosylated histo-blood group antigens are markers of breast cancer-initiating cells

et al. 2010

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Histo-blood group antigens CD173 (H2) and CD174 (Lewis Y) are known to be developmentally regulated carbohydrate antigens which are expressed to a varying degree on many human carcinomas. We hypothesized that they might represent markers of cancer-initiating cells (or cancer stem cells, CSC). In order to test this hypothesis, we examined the co-expression of CD173 and CD174 with stem cell markers CD44 and CD133 by flow cytometry analysis, immunocytochemistry, and immunohistochemistry on cell lines and tissue sections from breast cancer. In three breast cancer cell lines, the percentage of CD173(+)/CD44(+) cells ranged from 17% to >60% and of CD174(+)/CD44(+) from 21% to 57%. In breast cancer tissue sections from 15 patients, up to 50% of tumor cells simultaneously expressed CD173, CD174, and CD44 antigens. Co-expression of CD173 and CD174 with CD133 was also observed, but to a lesser percentage. Co-immunoprecipitation and sandwich ELISA experiments on breast cancer cell lines suggested that CD173 and CD174 are carried on the CD44 molecule. The results show that in these tissues CD173 (H2) and CD174 (LeY) are associated with CD44 expression, suggesting that these carbohydrate antigens are markers of cancer-initiating cells or of early progenitors of breast carcinomas.

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A new monoclonal antibody (A46-B/B10) highly specific for the blood group H type 2 epitope: Generation, epitope analysis, serological and histological evaluation

Cited by 13 publications

References 23 publications

Protein fucosylation regulates synapsin Ia/Ib expression and neuronal morphology in primary hippocampal neurons

Protein fucosylation regulates synapsin Ia/Ib expression and neuronal morphology in primary hippocampal neurons

Modifying Antibody Specificity by Chain Shuffling of V_H / V_L between Antibodies with Related Specificities

Co-expression of CD173 (H2) and CD174 (Lewis Y) with CD44 suggests that fucosylated histo-blood group antigens are markers of breast cancer-initiating cells

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A new monoclonal antibody (A46-B/B10) highly specific for the blood group H type 2 epitope: Generation, epitope analysis, serological and histological evaluation

Cited by 13 publications

References 23 publications

Protein fucosylation regulates synapsin Ia/Ib expression and neuronal morphology in primary hippocampal neurons

Protein fucosylation regulates synapsin Ia/Ib expression and neuronal morphology in primary hippocampal neurons

Modifying Antibody Specificity by Chain Shuffling of VH / VL between Antibodies with Related Specificities

Co-expression of CD173 (H2) and CD174 (Lewis Y) with CD44 suggests that fucosylated histo-blood group antigens are markers of breast cancer-initiating cells

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Modifying Antibody Specificity by Chain Shuffling of V_H / V_L between Antibodies with Related Specificities