Phosphorylated di-,tri-and tetra-saccharides ofβ-1,2-mannan antigen derived from Candida albicans(C.albicans)cell wall were synthesized and covalently conjugated with keyhole limpet hemocyanin(KLH)and human serum alb...Phosphorylated di-,tri-and tetra-saccharides ofβ-1,2-mannan antigen derived from Candida albicans(C.albicans)cell wall were synthesized and covalently conjugated with keyhole limpet hemocyanin(KLH)and human serum albumin(HSA)via a bifunctional linker under mild conditions.The semi-syntheticβ-1,2-mannoside–KLH conjugates were evaluated for the immunization of BALB/c mice.The ELISA results revealed that all three conjugates could elicit high levels of specific IgG antibodies and the acquired antisera could effectively identify theβ-1,2-mannan epitope.Furthermore,the immunofluorescence and flow cytometry assays also uncovered that the induced antibodies,especially that obtained from immunization withβ-1,2-mannotriose–KLH conjugate(1b),could bind well to fungi cell.Eventually,the structure–immunogenicity relationship analysis ofβ-mannan showed that the length of oligo-β-mannoses had a big impact on their immunogenicity andβ-1,2-mannotriose showed the strongest immunogenicity.The results suggested the great potential ofβ-1,2-mannotriose–KLH conjugate as an antifungal vaccine candidate.展开更多
The dynamic distribution of three different glycoconjugates in oocytes and follicle cells during the oogenesis of Atractomorpha sinensis were detected using biotin-labeled Peanut Agglutinin(PNA),Soy Bean Agglutinin(SB...The dynamic distribution of three different glycoconjugates in oocytes and follicle cells during the oogenesis of Atractomorpha sinensis were detected using biotin-labeled Peanut Agglutinin(PNA),Soy Bean Agglutinin(SBA)and Ulex Europaeus Agglutinin I(UEA-I)lectins.The results showed that during oogenesis there was no distribution of the UEA-I receptor.The receptors of PNA and SBA were found to be dependent on developmental stage and present different distribution patterns accordingly.The binding sites of the two lectins indicated the presence of different sugars(PNA for Galβ1,3GalNAc and SBA for GalNAc)and showed considerable variation during oogenesis.PNA and SBA receptors first appeared at the oocyte growth phase,the PNA receptors then disappeared gradually and the SBA receptors exhibited the greatest expression.At the early phase of yolk formation,PNA and SBA receptors were located just at the brim of ooplasm,which was the region of vitellin formation.However at the later phase of yolk formation,neither of the two receptors was detected.In the mature egg,PNA and SBA receptors were distributed again on the vitellin membrane and the eggshell.The two receptors were also widely distributed in the follicular cells,showing similar distribution variation to the oocytes.The results indicate that the change and modification of the two receptors may be greatly related to the growth of oocytes,the preparation for yolk formation,the differentiation of follicular cells and the maturation of oocytes.The glycoconjugates on the vitellin membrane probably play important roles in sperm and egg recognition.The two lectins bound moderately or strongly to the eggshell,which indicates that the eggshell of A.sinensis contains the GalNAc and GalβGalNAc glycoproteins.展开更多
The gut pathogen Enterocloster bolteae(E.bolteae)has been associated with autism spectrum disorder(ASD).The development of an E.bolteae vaccine to prevent gastrointestinal diseases,might be beneficial for understandin...The gut pathogen Enterocloster bolteae(E.bolteae)has been associated with autism spectrum disorder(ASD).The development of an E.bolteae vaccine to prevent gastrointestinal diseases,might be beneficial for understanding and treating ASD.Capsular polysaccharide(CPS)is a major virulence factor for E.bolteae.Based on an antigenicity evaluation of oligosaccharides associated with E.bolteae CPS and a structural revision of this carbohydrate antigen,two series of glycans including the D-Manp-D-Rhap type oligosaccharides 13-18 and the D-Ribp-D-Rhap type disaccharides 19-23 related to E.bolteae WAL-16351 CPS were prepared.The hydrogen-bond mediated glycosylation and conformational locking strategy facilitated the constructions of two 1,2-cis-β-glycosidic linkages.Glycan microarray analysis revealed that oligosaccharides 4,5,and 19 are recognized by antibodies in the anti-E.bolteae sera.The sera IgG antibodies induced by glycoconjugate 19-CRM197 recognize the CPS and bacteria specifically,whereas the IgG antibodies induced respectively by glycoconjugates 4-CRM197 and 5-CRM197 showed almost no binding to the CPS and bacteria.These results indicated that disaccharide 19 is a potential candidate for the development of E.bolteae vaccines.展开更多
基金financially supported by the Natural Science Foundation of China (Nos. 21502223, 81773580)
文摘Phosphorylated di-,tri-and tetra-saccharides ofβ-1,2-mannan antigen derived from Candida albicans(C.albicans)cell wall were synthesized and covalently conjugated with keyhole limpet hemocyanin(KLH)and human serum albumin(HSA)via a bifunctional linker under mild conditions.The semi-syntheticβ-1,2-mannoside–KLH conjugates were evaluated for the immunization of BALB/c mice.The ELISA results revealed that all three conjugates could elicit high levels of specific IgG antibodies and the acquired antisera could effectively identify theβ-1,2-mannan epitope.Furthermore,the immunofluorescence and flow cytometry assays also uncovered that the induced antibodies,especially that obtained from immunization withβ-1,2-mannotriose–KLH conjugate(1b),could bind well to fungi cell.Eventually,the structure–immunogenicity relationship analysis ofβ-mannan showed that the length of oligo-β-mannoses had a big impact on their immunogenicity andβ-1,2-mannotriose showed the strongest immunogenicity.The results suggested the great potential ofβ-1,2-mannotriose–KLH conjugate as an antifungal vaccine candidate.
文摘The dynamic distribution of three different glycoconjugates in oocytes and follicle cells during the oogenesis of Atractomorpha sinensis were detected using biotin-labeled Peanut Agglutinin(PNA),Soy Bean Agglutinin(SBA)and Ulex Europaeus Agglutinin I(UEA-I)lectins.The results showed that during oogenesis there was no distribution of the UEA-I receptor.The receptors of PNA and SBA were found to be dependent on developmental stage and present different distribution patterns accordingly.The binding sites of the two lectins indicated the presence of different sugars(PNA for Galβ1,3GalNAc and SBA for GalNAc)and showed considerable variation during oogenesis.PNA and SBA receptors first appeared at the oocyte growth phase,the PNA receptors then disappeared gradually and the SBA receptors exhibited the greatest expression.At the early phase of yolk formation,PNA and SBA receptors were located just at the brim of ooplasm,which was the region of vitellin formation.However at the later phase of yolk formation,neither of the two receptors was detected.In the mature egg,PNA and SBA receptors were distributed again on the vitellin membrane and the eggshell.The two receptors were also widely distributed in the follicular cells,showing similar distribution variation to the oocytes.The results indicate that the change and modification of the two receptors may be greatly related to the growth of oocytes,the preparation for yolk formation,the differentiation of follicular cells and the maturation of oocytes.The glycoconjugates on the vitellin membrane probably play important roles in sperm and egg recognition.The two lectins bound moderately or strongly to the eggshell,which indicates that the eggshell of A.sinensis contains the GalNAc and GalβGalNAc glycoproteins.
基金the National Natural Science Foundation of China(Nos.22325803,22277042,22177041,22207042,22107037)the Max Planck Society International Partner Group Program,the China Scholarship Council(CSC)the Fundamental Research Funds for the Central Universities for funding。
文摘The gut pathogen Enterocloster bolteae(E.bolteae)has been associated with autism spectrum disorder(ASD).The development of an E.bolteae vaccine to prevent gastrointestinal diseases,might be beneficial for understanding and treating ASD.Capsular polysaccharide(CPS)is a major virulence factor for E.bolteae.Based on an antigenicity evaluation of oligosaccharides associated with E.bolteae CPS and a structural revision of this carbohydrate antigen,two series of glycans including the D-Manp-D-Rhap type oligosaccharides 13-18 and the D-Ribp-D-Rhap type disaccharides 19-23 related to E.bolteae WAL-16351 CPS were prepared.The hydrogen-bond mediated glycosylation and conformational locking strategy facilitated the constructions of two 1,2-cis-β-glycosidic linkages.Glycan microarray analysis revealed that oligosaccharides 4,5,and 19 are recognized by antibodies in the anti-E.bolteae sera.The sera IgG antibodies induced by glycoconjugate 19-CRM197 recognize the CPS and bacteria specifically,whereas the IgG antibodies induced respectively by glycoconjugates 4-CRM197 and 5-CRM197 showed almost no binding to the CPS and bacteria.These results indicated that disaccharide 19 is a potential candidate for the development of E.bolteae vaccines.
