A structurally diversified linker enhances the immune response to a small carbohydrate hapten

Lipiński, Tomasz; Luu, Thanh; Kitov, Pavel I.; Szpacenko, Adam; Bundle, David R.

doi:10.1007/s10719-011-9331-8

Cited by 27 publications

(31 citation statements)

References 41 publications

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“…Interspecies (goat vs. mouse, rabbit vs. mouse) as well as mouse inter-strain differences in the ability to mount a glycan-specific antibody response to both native glycoproteins and synthetic glycoconjugates have been previously noted in the literature 17,32–37 . One explanation for the observed differences between rabbits and mice could be a differentially tolerized immune repertoire due to differences between murine and rabbit milk oligosaccharide composition.…”

Section: Discussionmentioning

confidence: 90%

Intact Reducing Glycan Promotes the Specific Immune Response to Lacto-N-neotetraose-BSA Neoglycoconjugates

et al. 2015

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The mammalian immune system responds to eukaryotic glycan antigens during infections, cancer, and autoimmune disorders, but the immunological bases for such responses are unclear. Conjugate vaccines containing bacterial polysaccharides linked to carrier proteins (neoglycoconjugates) have proven successful, but these often contain repeating epitopes and the reducing end of the glycan is less important, unlike typical glycan determinants in eukaryotes, which are shorter in length and may include the reducing end. Here we have compared the effects of two linkage methods, one that opens the ring at the reducing end of the glycan, and one that leaves the reducing end closed, on the glycan specificity of the vaccine response in rabbits and mice. We immunized rabbits and mice with bovine serum albumin (BSA) conjugates of synthetic open- and closed-ring forms (OR versus CR) of a simple tetrasaccharide lacto-N-neo-tetraose (LNnT, Galβ1-4GlcNAcβ1-3Galβ1-4Glc), and tested reactivity to the immunogens and several related glycans in both OR and CR versions on glycan microarrays. We found that in rabbits the immune response to the CR conjugate was directed toward the glycan, whereas the OR conjugate elicited antibodies to the reducing end of the glycan and linker region but not specifically to the glycan itself. Unexpectedly, mice did not generate a glycan-specific response to the CR conjugate. Our findings indicate that the reducing end of the sugar is crucial for generation of a glycan-specific response to some eukaryotic vaccine epitopes, and that there are species-specific differences in the ability to make a glycan-specific response to some glycoconjugates. These findings warrant further investigation with regard to rational design of glycoconjugate vaccines.

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Section: Discussionmentioning

confidence: 90%

Intact Reducing Glycan Promotes the Specific Immune Response to Lacto-N-neotetraose-BSA Neoglycoconjugates

et al. 2015

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“…This was in sharp contrast to the poor immunogenicity of b-mannotriose-tetanus toxoid conjugate in mice (32,33,37). These observations prompted us to consider DC targeting as a means to enhance the antigenicity of the b-mannan-tetanus toxoid conjugate.…”

Section: Discussionmentioning

confidence: 99%

“…This vaccine induced high titers of b-mannan-specific Abs, in the range of 20,000-80,000, after only two injections using alum as adjuvant (32). Disappointingly, the same vaccine given to mice produced corresponding titers of 10,000 or lower, and these Ab levels were not significantly improved by more aggressive immunization protocols that employed powerful adjuvants such as CFA (33).…”

mentioning

confidence: 99%

“…A conjugate vaccine that is composed of synthetic b-mannan trisaccharide linked to tetanus toxoid (11,30,31) achieved significant reductions in fungal counts in an immunocompromised rabbit model of disseminated candidiasis (32). This vaccine induced high titers of b-mannan-specific Abs, in the range of 20,000-80,000, after only two injections using alum as adjuvant (32).…”

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confidence: 99%

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Enhanced Immunogenicity of a Tricomponent Mannan Tetanus Toxoid Conjugate Vaccine Targeted to Dendritic Cells via Dectin-1 by Incorporating β-Glucan

Lipiński

Fitieh

St‐Pierre

et al. 2013

The Journal of Immunology

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In a previous attempt to generate a protective vaccine against Candida albicans, a β-mannan tetanus toxoid conjugate showed poor immunogenicity in mice. To improve the specific activation toward the fungal pathogen, we aimed to target Dectin-1, a pattern-recognition receptor expressed on monocytes, macrophages, and dendritic cells. Laminarin, a β-glucan ligand of Dectin-1, was incorporated into the original β-mannan tetanus toxoid conjugate providing a tricomponent conjugate vaccine. A macrophage cell line expressing Dectin-1 was employed to show binding and activation of Dectin-1 signal transduction pathway by the β-glucan–containing vaccine. Ligand binding to Dectin-1 resulted in the following: 1) activation of Src family kinases and Syk revealed by their recruitment and phosphorylation in the vicinity of bound conjugate and 2) translocation of NF-κB to the nucleus. Treatment of immature bone marrow–derived dendritic cells (BMDCs) with tricomponent or control vaccine confirmed that the β-glucan–containing vaccine exerted its enhanced activity by virtue of dendritic cell targeting and uptake. Immature primary cells stimulated by the tricomponent vaccine, but not the β-mannan tetanus toxoid vaccine, showed activation of BMDCs. Moreover, treated BMDCs secreted increased levels of several cytokines, including TGF-β and IL-6, which are known activators of Th17 cells. Immunization of mice with the novel type of vaccine resulted in improved immune response manifested by high titers of Ab recognizing C. albicans β-mannan Ag. Vaccine containing laminarin also affected distribution of IgG subclasses, showing that vaccine targeting to Dectin-1 receptor can benefit from augmentation and immunomodulation of the immune response.

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“…Nevertheless, this competition between hapten and linkers, especially for conjugates utilising smaller oligosaccharides as the carbohydrate component, is going to effect the maturation of the immune response to the desired carbohydrate target. A recent paper by Lipinski et al [32] has introduced a novel alternative conjugation strategy, by utilising stereochemically variable linker molecules to suppress the immune response maturation to the linker moieties. The present study has therefore clearly confirmed and extended the potential of the inner core LPS as a vaccine candidate to combat disease caused by the significant veterinary pathogen M. haemolytica.…”

Section: Discussionmentioning

confidence: 99%

Investigating the candidacy of lipopolysaccharide-based glycoconjugates as vaccines to combat Mannheimia haemolytica

et al. 2011

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Inner core lipopolysaccharide (LPS) has been shown to be conserved in the majority of veterinary strains from the species Mannheimia haemolytica, Actinobacillus pleuropneumoniae and Pasteurella multocida and as such is being considered as a possible vaccine antigen. The proof-in-principle that a LPS-based antigen could be considered as a vaccine candidate has been demonstrated from studies with monoclonal antibodies raised to the inner core LPS of Mannheimia haemolytica, which were shown to be both bactericidal and protective in a mouse model of disease. In this study we confirm and extend the candidacy of the inner core LPS by demonstrating that it is possible to elicit functional antibodies against Mannheimia haemolytica wild-type strains following immunisation of rabbits with glycoconjugates elaborating the conserved inner core LPS antigen. The present study describes a conjugation strategy that uses amidases produced by Dictyostelium discoideum, targeting the amino functionality created by the amidase activity as the attachment point on the LPS molecule. To protect the amino functionality on the phosphoethanolamine (PEtn) residue of the inner core, we developed a novel blocking and unblocking strategy with t-butyl oxycarbonyl. A maleimide-thiol linker strategy with the thiol linker on the carboxyl residues of the carrier protein and the maleimide linker on the carbohydrate resulted in a high loading of carbohydrates per carrier protein. Immunisation derived antisera from rabbits recognised fully extended Mannheimia haemolytica LPS and whole cells from serotypes 1 and 2, despite a somewhat immunodominant response to the linkers also being observed. Moreover, bactericidal activity was demonstrated to a strain elaborating the immunising carbohydrate antigen and crucially to wild-type cells of serotypes 1 and 2, thus further supporting the consideration of inner core LPS as a potential vaccine antigen to combat disease caused by Mannheimia haemolytica.

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A structurally diversified linker enhances the immune response to a small carbohydrate hapten

Cited by 27 publications

References 41 publications

Intact Reducing Glycan Promotes the Specific Immune Response to Lacto-N-neotetraose-BSA Neoglycoconjugates

Intact Reducing Glycan Promotes the Specific Immune Response to Lacto-N-neotetraose-BSA Neoglycoconjugates

Enhanced Immunogenicity of a Tricomponent Mannan Tetanus Toxoid Conjugate Vaccine Targeted to Dendritic Cells via Dectin-1 by Incorporating β-Glucan

Investigating the candidacy of lipopolysaccharide-based glycoconjugates as vaccines to combat Mannheimia haemolytica

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