RESUMO
Glycoconjugate vaccines are based on chemical conjugation of pathogen-associated carbohydrates with immunogenic carrier proteins and are considered a very cost-effective way to prevent infections. Most of the licensed glycoconjugate vaccines are composed of saccharide antigens extracted from bacterial sources. However, synthetic oligosaccharide antigens have become a promising alternative to natural polysaccharides with the advantage of being well-defined structures providing homogeneous conjugates. Haemophilus influenzae (Hi) is responsible for a number of severe diseases. In recent years, an increasing rate of invasive infections caused by Hi serotype a (Hia) raised some concern, because no vaccine targeting Hia is currently available. The capsular polysaccharide (CPS) of Hia is constituted by phosphodiester-linked 4-ß-d-glucose-(1â4)-d-ribitol-5-(PO4â) repeating units and is the antigen for protein-conjugated polysaccharide vaccines. To investigate the antigenic potential of the CPS from Hia, we synthesized related saccharide fragments containing up to five repeating units. Following the synthetic optimization of the needed disaccharide building blocks, they were assembled using the phosphoramidite approach for the installation of the phosphodiester linkages. The resulting CPS-based Hia oligomers were conjugated to CRM197 carrier protein and evaluated inâ vivo for their immunogenic potential, showing that all glycoconjugates were capable of raising antibodies recognizing Hia synthetic fragments.
Assuntos
Glicoconjugados , Haemophilus influenzae , Glicoconjugados/química , Glicoconjugados/imunologia , Glicoconjugados/síntese química , Haemophilus influenzae/imunologia , Haemophilus influenzae/química , Animais , Vacinas Conjugadas/química , Vacinas Conjugadas/imunologia , Camundongos , Vacinas Anti-Haemophilus/imunologia , Vacinas Anti-Haemophilus/química , Proteínas de Bactérias/imunologia , Proteínas de Bactérias/química , Infecções por Haemophilus/prevenção & controle , Infecções por Haemophilus/imunologiaRESUMO
Reported herein is a new HPLC-based automated synthesizer (HPLC-A) capable of a temperature-controlled synthesis and purification of carbohydrates. The developed platform allows to perform various protecting group manipulations as well as the synthesis of O- and N-glycosides. A fully automated synthesis and purification was showcased in application to different carbohydrate derivatives including glycosides, oligosaccharides, glycopeptides, glycolipids, and nucleosides.
Assuntos
Carboidratos , Oligossacarídeos , Cromatografia Líquida de Alta Pressão , Carboidratos/química , Oligossacarídeos/síntese química , Oligossacarídeos/química , Glicosídeos/química , Glicosídeos/síntese química , Glicolipídeos/química , Glicolipídeos/síntese química , Glicolipídeos/isolamento & purificação , Glicopeptídeos/química , Glicopeptídeos/síntese química , Nucleosídeos/química , Nucleosídeos/síntese química , Automação , TemperaturaRESUMO
Group A Carbohydrate (GAC), conjugated to an appropriate carrier protein, has been proposed as an attractive vaccine candidate against Group A Streptococcus infections. Native GAC consists of a polyrhamnose (polyRha) backbone with N-acetylglucosamine (GlcNAc) at every second rhamnose residue. Both native GAC and the polyRha backbone have been proposed as vaccine components. Here, chemical synthesis and glycoengineering were used to generate a panel of different length GAC and polyrhamnose fragments. Biochemical analyses were performed confirming that the epitope motif of GAC is composed of GlcNAc in the context of the polyrhamnose backbone. Conjugates from GAC isolated and purified from a bacterial strain and polyRha genetically expressed in E. coli and with similar molecular size to GAC were compared in different animal models. The GAC conjugate elicited higher anti-GAC IgG levels with stronger binding capacity to Group A Streptococcus strains than the polyRha one, both in mice and in rabbits. This work contributes to the development of a vaccine against Group A Streptococcus suggesting GAC as preferable saccharide antigen to include in the vaccine.
Assuntos
Acetilglucosamina , Vacinas , Camundongos , Animais , Coelhos , Acetilglucosamina/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Carboidratos , Streptococcus pyogenes/metabolismo , Glicoconjugados/metabolismoRESUMO
Ultra-small gold nanoparticles (UAuNPs) are extremely interesting for applications in nanomedicine thanks to their good stability, biocompatibility, long circulation time and efficient clearance pathways. UAuNPs engineered with glycans (Glyco-UAuNPs) emerged as excellent platforms for many applications since the multiple copies of glycans can mimic the multivalent effect of glycoside clusters. Herein, we unravel a straightforward photo-induced synthesis of Glyco-UAuNPs based on a reliable and robust microfluidic approach. The synthesis occurs at room temperature avoiding the use of any further chemical reductant, templating agents or co-solvents. Exploiting 1 Hâ NMR spectroscopy, we showed that the amount of thiol-ligand exposed on the UAuNPs is linearly correlated to the ligand concentration in the initial mixture. The results pave the way towards the development of a programmable synthetic approach, enabling an accurate design of the engineered UAuNPs or smart hybrid nano-systems.
Assuntos
Nanopartículas Metálicas , Nanopartículas , Ouro/química , Nanopartículas Metálicas/química , Microfluídica , Ligantes , Nanopartículas/química , Polissacarídeos/químicaRESUMO
Vaccination is one of the greatest achievements in biomedical research preventing death and morbidity in many infectious diseases through the induction of pathogen-specific humoral and cellular immune responses. Currently, no effective vaccines are available for pathogens with a highly variable antigenic load, such as the human immunodeficiency virus or to induce cellular T-cell immunity in the fight against cancer. The recent SARS-CoV-2 outbreak has reinforced the relevance of designing smart therapeutic vaccine modalities to ensure public health. Indeed, academic and private companies have ongoing joint efforts to develop novel vaccine prototypes for this virus. Many pathogens are covered by a dense glycan-coat, which form an attractive target for vaccine development. Moreover, many tumor types are characterized by altered glycosylation profiles that are known as "tumor-associated carbohydrate antigens". Unfortunately, glycans do not provoke a vigorous immune response and generally serve as T-cell-independent antigens, not eliciting protective immunoglobulin G responses nor inducing immunological memory. A close and continuous crosstalk between glycochemists and glycoimmunologists is essential for the successful development of efficient immune modulators. It is clear that this is a key point for the discovery of novel approaches, which could significantly improve our understanding of the immune system. In this review, we discuss the latest advancements in development of vaccines against glycan epitopes to gain selective immune responses and to provide an overview on the role of different immunogenic constructs in improving glycovaccine efficacy.
Assuntos
COVID-19 , Neoplasias , Vacinas , COVID-19/prevenção & controle , Glicoconjugados/uso terapêutico , Humanos , Neoplasias/prevenção & controle , Polissacarídeos/uso terapêutico , SARS-CoV-2RESUMO
Vaccination represents the most effective way to prevent invasive pneumococcal diseases. The glycoconjugate vaccines licensed so far are obtained from capsular polysaccharides (CPSs) of the most virulent serotypes. Protection is largely limited to the specific vaccine serotypes, and the continuous need for broader coverage to control the outbreak of emerging serotypes is pushing the development of new vaccine candidates. Indeed, the development of efficacious vaccine formulation is complicated by the high number of bacterial serotypes with different CPSs. In this context, to simplify vaccine composition, we propose the design of new saccharide fragments containing chemical structures shared by different serotypes as cross-reactive and potentially cross-protective common antigens. In particular, we focused on Streptococcus pneumoniae (Sp) 19A and 19F. The CPS repeating units of Sp 19F and 19A are very similar and share a common structure, the disaccharide ManNAc-ß-(1â4)-Glc (A-B). Herein, we describe the synthesis of a small library of compounds containing different combinations of the common 19F/19A disaccharide. The six new compounds were tested with a glycan array to evaluate their recognition by antibodies in reference group 19 antisera and factor reference antisera (reacting against 19F or 19A). The disaccharide A-B, phosphorylated at the upstream end, emerged as a hit from the glycan array screening because it is strongly recognized by the group 19 antisera and by the 19F and 19A factor antisera, with similar intensity compared with the CPSs used as controls. Our data give a strong indication that the phosphorylated disaccharide A-B can be considered a common epitope among different Sp 19 serotypes.
Assuntos
Epitopos/química , Glicoconjugados/análise , Proteínas Imobilizadas/química , Polissacarídeos Bacterianos/análise , Anticorpos/química , Técnicas Biossensoriais , Reações Cruzadas , Glicoconjugados/metabolismo , Hexosaminas/química , Polissacarídeos Bacterianos/metabolismo , Sorogrupo , Soro/química , Espectrometria de Fluorescência , Streptococcus pneumoniae/metabolismo , Propriedades de SuperfícieRESUMO
Glycan structures are common posttranslational modifications of proteins, which serve multiple important structural roles (for instance in protein folding), but also are crucial participants in cell-cell communications and in the regulation of immune responses. Through the interaction with glycan-binding receptors, glycans are able to affect the activation status of antigen-presenting cells, leading either to induction of pro-inflammatory responses or to suppression of immunity and instigation of immune tolerance. This unique feature of glycans has attracted the interest and spurred collaborations of glyco-chemists and glyco-immunologists to develop glycan-based tools as potential therapeutic approaches in the fight against diseases such as cancer and autoimmune conditions. In this review, we highlight emerging advances in this field, and in particular, we discuss on how glycan-modified conjugates or glycoengineered cells can be employed as targeting devices to direct tumor antigens to lectin receptors on antigen-presenting cells, like dendritic cells. In addition, we address how glycan-based nanoparticles can act as delivery platforms to enhance immune responses. Finally, we discuss some of the latest developments in glycan-based therapies, including chimeric antigen receptor (CAR)-T cells to achieve targeting of tumor-associated glycan-specific epitopes, as well as the use of glycan moieties to suppress ongoing immune responses, especially in the context of autoimmunity.
Assuntos
Autoimunidade/imunologia , Polissacarídeos/imunologia , Receptores de Antígenos Quiméricos/imunologia , Linfócitos T/imunologia , Animais , Comunicação Celular/imunologia , Humanos , Nanopartículas/química , Polissacarídeos/química , Processamento de Proteína Pós-TraducionalRESUMO
Lectins are involved in a wide range of carbohydrate mediated recognition processes. Therefore, the availability of highly performant fluorescent tools tailored for lectin targeting and able to efficiently track events related to such key targets is in high demand. We report here on the synthesis of the glyco-BODIPYs 1 and 2, based on the efficient combination of a Heck-like cross coupling and a Knoevenagel condensation, which revealed efficient in addressing lectins. In particular, glyco-BODIPY 1 has two glycosidase stable C-mannose residues, which act as DC-SIGN (dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin) targeting modules. By using live-cell fluorescence microscopy, we proved that BODIPY-mannose 1 was efficiently taken up by immune cells expressing DC-SIGN receptors. Super-resolution stimulated emission depletion (STED) microscopy further revealed that the internalized 1 localized in membranes of endosomes, proving that 1 is a reliable tool also in STED applications. Of note, glyco-BODIPY 1 contains an aryl-azido group, which allows further functionalization of the glycoprobe with bioactive molecules, thus paving the way for the use of 1 for tracking lectin-mediated cell internalization in diverse biological settings.
Assuntos
Compostos de Boro/química , Moléculas de Adesão Celular/análise , Lectinas Tipo C/análise , Receptores de Superfície Celular/análise , Compostos de Boro/síntese química , Linhagem Celular , Relação Dose-Resposta a Droga , Glucose/química , Voluntários Saudáveis , Humanos , Manose/química , Estrutura Molecular , Relação Estrutura-AtividadeRESUMO
Carbohydrates are ubiquitous molecules expressed on the surface of nearly all living cells, and their interaction with carbohydrate-binding proteins is critical to many immunobiological processes. Carbohydrates are utilized as antigens in many licensed vaccines against bacterial pathogens. More recently, they have also been considered as adjuvants. Interestingly, unlike other types of vaccines, adjuvants have improved immune response to carbohydrate-based vaccine in humans only in a few cases. Furthermore, despite the discovery of many new adjuvants in the last years, aluminum salts, when needed, remain the only authorized adjuvant for carbohydrate-based vaccines. In this review, we highlight historical and recent advances on the use of glycans either as vaccine antigens or adjuvants, and we review the use of currently available adjuvants to improve the efficacy of carbohydrate-based vaccines. A better understanding of the mechanism of carbohydrate interaction with innate and adaptive immune cells will benefit the design of a new generation of glycan-based vaccines and of immunomodulators to fight both longstanding and emerging diseases.
Assuntos
Doenças Transmissíveis , Vacinas , Adjuvantes Imunológicos , Antígenos , Carboidratos , HumanosRESUMO
A Correction to this paper has been published: https://doi.org/10.1038/s41467-020-20120-4.
RESUMO
Neisseria meningitidis serogroup A capsular polysaccharide (MenA CPS) consists of (1 â 6)-2-acetamido-2-deoxy-α-D-mannopyranosyl phosphate repeating units, O-acetylated at position C3 or C4. Glycomimetics appear attractive to overcome the CPS intrinsic lability in physiological media, due to cleavage of the phosphodiester bridge, and to develop a stable vaccine with longer shelf life in liquid formulation. Here, we generate a series of non-acetylated carbaMenA oligomers which are proven more stable than the CPS. An octamer (DP8) inhibits the binding of a MenA specific bactericidal mAb and polyclonal serum to the CPS, and is selected for further in vivo testing. However, its CRM197 conjugate raises murine antibodies towards the non-acetylated CPS backbone, but not the natural acetylated form. Accordingly, random O-acetylation of the DP8 is performed, resulting in a structure (Ac-carbaMenA) showing improved inhibition of anti-MenA CPS antibody binding and, after conjugation to CRM197, eliciting anti-MenA protective murine antibodies, comparably to the vaccine benchmark.
Assuntos
Glicoconjugados/síntese química , Neisseria meningitidis Sorogrupo A/imunologia , Polissacarídeos Bacterianos/síntese química , Vacinas Conjugadas , Animais , Anticorpos Antibacterianos/análise , Anticorpos Neutralizantes/química , Cápsulas Bacterianas/imunologia , Biomimética/métodos , Glicoconjugados/imunologia , Camundongos , Neisseria meningitidis Sorogrupo A/química , Neisseria meningitidis Sorogrupo A/efeitos dos fármacos , Polissacarídeos Bacterianos/química , Polissacarídeos Bacterianos/imunologia , Vacinas Conjugadas/química , Vacinas Conjugadas/microbiologiaRESUMO
The development of novel delivery systems capable of enhancing the antibody binding affinity and immunoactivity of short length saccharide antigens is at the forefront of modern medicine. In this regard, gold nanoparticles (AuNPs) raised great interest as promising nano-vaccine platform, as they do not interfere with the desired immune response and their surface can be easily functionalized, enabling the antigen multivalent presentation. In addition, the nanoparticles morphology can have a great impact on their biological properties. Gram-positive Group A Streptococcus (GAS) is a bacterium responsible for many infections and represents a priority healthcare concern, but a universal vaccine is still unavailable. Since all the GAS strains have a cell wall characterized by a common polyrhamnose backbone, this can be employed as alternative antigen to develop an anti-GAS vaccine. Herein, we present the synthesis of two oligorhamnoside fragments and their corresponding oligorhamnoside-AuNPs, designed with two different morphologies. By competitive ELISA we assessed that both symmetric and anisotropic oligorhamnan nanoparticles inhibit the binding of specific polyclonal serum much better than the unconjugated oligosaccharides.
Assuntos
Anticorpos/imunologia , Ouro/química , Nanopartículas Metálicas/química , Oligorribonucleotídeos/imunologia , Streptococcus/química , Anticorpos/química , Configuração de Carboidratos , Ouro/imunologia , Oligorribonucleotídeos/síntese química , Oligorribonucleotídeos/química , Streptococcus/imunologiaRESUMO
Since their discovery, therapeutic or prophylactic vaccines represent a promising option to prevent or cure infections and other pathologies, such as cancer or autoimmune disorders. More recently, among a number of nanomaterials, gold nanoparticles (AuNPs) have emerged as novel tools for vaccine developments, thanks to their inherent ability to tune and upregulate immune response. Moreover, owing to their features, AuNPs can exert optimal actions both as delivery systems and as adjuvants. Notwithstanding the potential huge impact in vaccinology, some challenges remain before AuNPs in vaccine formulations can be translated into the clinic. The current review provides an updated overview of the most recent and effective application of gold nanoparticles as efficient means to develop a new generation of vaccine.
Assuntos
Nanopartículas Metálicas , Vacinas , Adjuvantes Imunológicos , Ouro , Desenvolvimento de VacinasRESUMO
Streptococcus pneumoniae (SP) is a common human pathogen associated with a broad spectrum of diseases and it is still a leading cause of mortality and morbidity worldwide, especially in children. Moreover, SP is increasingly associated with drug resistance. Vaccination against the pathogen may thus represent an important strategy to overcome its threats to human health. In this context, revealing the molecular determinants of SP immunoreactivity may be relevant for the development of novel molecules with therapeutic perspectives as vaccine components. Serogroup 19 comprises the immune-cross reactive types 19F, 19A, 19B and 19C and it accounts for a high percentage of invasive pneumococcal diseases, mainly caused by serotypes 19F and 19A. Herein, we report the synthesis and biological evaluation of an aminopropyl derivative of the trisaccharide repeating unit of SP 19A. We compare two different synthetic strategies, based on different disconnections between the three monosaccharides which make up the final trisaccharide, to define the best approach for the preparation of the trisaccharide. Synthetic accessibility to the trisaccharide repeating unit lays the basis for the development of more complex biopolymer as well as saccharide conjugates. We also evaluate the binding affinity of the trisaccharide for anti-19A and anti-19F sera and discuss the relationship between the chemical properties of the trisaccharide unit and biological activity.
Assuntos
Trissacarídeos/síntese química , Trissacarídeos/imunologia , Animais , Anticorpos/imunologia , Sequência de Carboidratos , Bovinos , Reações Cruzadas , Polissacarídeos Bacterianos/química , Coelhos , Estereoisomerismo , Streptococcus pneumoniae/química , Trissacarídeos/sangueRESUMO
Neisseria meningitidis serogroup A (MenA) is an aerobic diplococcal Gram-negative bacterium responsible for epidemic meningitis disease. Its capsular polysaccharide (CPS) has been identified as the primary virulence factor of MenA. This polysaccharide suffers from chemical lability in water. Thus, the design and synthesis of novel and hydrolytically stable structural analogues of MenA CPS may provide additional tools for the development of therapies against this disease. In this context, the structural features of the natural phosphorylated monomer have been analyzed and compared to those of its carba-analogue, where the endocyclic oxygen has been replaced by a methylene moiety. The lowest energy geometries of the different molecules have been calculated using a combination of quantum mechanical techniques and molecular dynamics simulations. The predicted results have been compared and validated using NMR experiments. The results indicate that the more stable designed glycomimetics may adopt the conformation adopted by the natural monomer, although they display a wider flexibility around the torsional degrees of freedom.
RESUMO
During the last decade there has been a growing interest in glycoimmunology, a relatively new research field dealing with the specific interactions of carbohydrates with the immune system. Pathogens' cell surfaces are covered by a thick layer of oligo- and polysaccharides that are crucial virulence factors, as they mediate receptors binding on host cells for initial adhesion and organism invasion. Since in most cases these saccharide structures are uniquely exposed on the pathogen surface, they represent attractive targets for vaccine design. Polysaccharides isolated from cell walls of microorganisms and chemically conjugated to immunogenic proteins have been used as antigens for vaccine development for a range of infectious diseases. However, several challenges are associated with carbohydrate antigens purified from natural sources, such as their difficult characterization and heterogeneous composition. Consequently, glycoconjugates with chemically well-defined structures, that are able to confer highly reproducible biological properties and a better safety profile, are at the forefront of vaccine development. Following on from our previous review on the subject, in the present account we specifically focus on the most recent advances in the synthesis and preliminary immunological evaluation of next generation glycoconjugate vaccines designed to target bacterial and fungal infections that have been reported in the literature since 2011.
Assuntos
Infecções Bacterianas/prevenção & controle , Vacinas Bacterianas , Glicoconjugados , Micoses/prevenção & controle , Fatores de Virulência , Animais , Infecções Bacterianas/imunologia , Vacinas Bacterianas/química , Vacinas Bacterianas/imunologia , Vacinas Fúngicas/química , Vacinas Fúngicas/imunologia , Glicoconjugados/química , Glicoconjugados/imunologia , Humanos , Micoses/imunologia , Fatores de Virulência/química , Fatores de Virulência/imunologiaRESUMO
Biological fate and toxicity of nanoparticles (NPs) are connected to the interaction between NPs and the protein corona (PC) spontaneously forming around NPs in biological matrixes. PC is a dynamic entity that confers biological identity to NPs. In this work, fluorescence cross-correlation spectroscopy (FCCS) is used to study the impact of specific interactions between the NP surface and proteins on the intracellular fate of PC. The stability of the PC formed around glucosamide-functionalized Au-NPs from ConcanavalinA (ConA) or Bovine Serum Albumin (BSA) is characterized by FCCS. The NPs show higher affinity for ConA and competitive assays show that ConA easily exchanges BSA. A549 cells are exposed to glucosamide-functionalized Au-NPs with preformed ConA and BSA PCs. Intracellularly the frequency of cross-correlation for Au NPs with ConA PC remains constant to a 70% value until 24 h while for BSA it decreases to a 15% during the same period. FCCS measurements in several locations in the cell point out a different level of aggregation for the NPs with either ConA or BSA PCs. Our results show that the affinity of NPs functionalized with a ligand with affinity for a specific protein in bulk is retained intracellularly influencing NP fate and translocation.
Assuntos
Concanavalina A/química , Glucosamina/química , Ouro/química , Nanopartículas Metálicas/química , Soroalbumina Bovina/química , Células A549 , Animais , Ligação Competitiva , Bovinos , Concanavalina A/metabolismo , Glucosamina/metabolismo , Ouro/metabolismo , Humanos , Nanopartículas Metálicas/ultraestrutura , Microscopia Confocal , Microscopia Eletrônica de Transmissão , Ligação Proteica , Soroalbumina Bovina/metabolismo , Espectrometria de Fluorescência , Propriedades de SuperfícieRESUMO
Studies on the polymerization mode of Neisseria meningitidis serogroup X capsular polymerase CsxA recently identified a truncated construct that can be immobilized and used for length controlled on-column production of oligosaccharides. Here, we combined the use of a synthetic acceptor bearing an appendix for carrier protein conjugation and the on-column process to a novel chemo-enzymatic strategy. After protein coupling of the size optimized oligosaccharide produced by the one-pot elongation procedure, we obtained a more homogeneous glycoconjugate compared to the one previously described starting from the natural polysaccharide. Mice immunized with the conjugated fully synthetic oligomer elicited functional antibodies comparable to controls immunized with the current benchmark MenX glycoconjugates prepared from the natural capsule polymer or from fragments of it enzymatically elongated. This pathogen-free technology allows the fast total in vitro construction of predefined bacterial polysaccharide fragments. Compared to conventional synthetic protocols, the procedure is more expeditious and drastically reduces the number of purification steps to achieve the oligomers. Furthermore, the presence of a linker for conjugation in the synthetic acceptor minimizes manipulations on the enzymatically produced glycan prior to protein conjugation. This approach enriches the methods for fast construction of complex bacterial carbohydrates.
Assuntos
Glicoconjugados/síntese química , Neisseria meningitidis/imunologia , Sorogrupo , Vacinas Conjugadas , Imunidade Adaptativa , Animais , Glicoconjugados/imunologia , Camundongos , Polissacarídeos Bacterianos/síntese químicaRESUMO
Star-shaped gold nanoparticles (StarAuNPs) are extremely attractive nanomaterials, characterized by localized surface plasmon resonance which could be potentially employed in a large number of applications. However, the lack of a reliable and reproducible synthetic protocols for the production of StarAuNPs is the major limitation to their spreading. For the first time, here we present a robust protocol to manufacture reproducible StarAuNPs by exploiting a fluidic approach. Star-shaped AuNPs have been synthesized by means of a seed-less protocol, employing ascorbic acid as reducing agent at room temperature. Moreover, the versatility of the bench-top microfluidic protocol has been exploited to afford hydrophilic, hydrophobic and solid-supported engineered StarAuNPs, by avoiding intermediate NP purifications.
RESUMO
AIM: Nanotechnology-based fully synthetic carbohydrate vaccines are promising alternatives to classic polysaccharide/protein conjugate vaccines. We have prepared gold glyco-nanoparticles (GNP) bearing two synthetic carbohydrate antigens related to serotypes 19F and 14 of Streptococcus pneumoniae and evaluated their immunogenicity in vivo. RESULTS: A tetrasaccharide fragment of serotype 14 (Tetra-14), a trisaccharide fragment of serotype 19F (Tri-19F), a T-helper peptide and d -glucose were loaded onto GNP in different ratios. Mice immunization showed that the concomitant presence of Tri-19F and Tetra-14 on the same nanoparticle critically enhanced the titers of specific IgG antibodies toward type 14 polysaccharide compared with GNP exclusively displaying Tetra-14, while no IgG antibodies against type 19F polysaccharide were elicited. CONCLUSION: This work is a step forward toward synthetic nanosystems combining carbohydrate antigens and immunogenic peptides as potential carbohydrate-based vaccines.