Elucidating the Structural Requirements for Next-Gen Glycoconjugate Vaccines

阐明下一代糖复合疫苗的结构要求

• 批准号: 10084269
• 负责人: Dennis L. Kasper
• 金额: $ 60.46万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-10 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10084269
• 关键词: Affinity; Antibodies; Antibody Affinity; Antibody Avidity; Antibody Response; Antibody titer measurement; Antibody-mediated protection; Antigen-Presenting Cells; Antigens; Attention; Avidity; B-Lymphocytes; B-cell receptor repertoire sequencing; Binding; Biological Markers; CD4 Positive T Lymphocytes; Carbohydrates; Carrier Proteins; Cells; Characteristics; Complex; Crystallization; Data; Disease; Epitopes; Evaluation; Event; Fab Immunoglobulins; Generations; Genetic Transcription; Glycoconjugates; Glycopeptides; Glycoproteins; Health Benefit; Helper-Inducer T-Lymphocyte; Histocompatibility Antigens Class II; Hybridomas; Immune; Immune response; Immunity; Immunization; Immunize; Immunoglobulin G; Immunologics; Link; Mediating; Memory B-Lymphocyte; Modeling; Molecular Weight; Monoclonal Antibodies; Oligosaccharides; Peptide Vaccines; Peptides; Polysaccharides; Population; Production; Proteins; Role; Site; Specificity; Streptococcus Group B; Structure; Surface; T-Lymphocyte; Tularemia; Vaccines; Work; adaptive immune response; alpha-beta T-Cell Receptor; base; design; high risk; insight; mouse model; pathogen; response; screening; sugar; vaccine-induced antibodies

Elucidating the Structural Requirements for Next-Gen Glycoconjugate Vaccines Project Summary Glycoconjugate vaccines provide enormous health benefits globally, although they have been less successful in some populations at high risk for developing disease. In most cases, conjugation of the sugar antigen to the carrier protein has been done empirically with little attention on variables critical for the immune response. Recent findings from our lab offer a new and rational explanation for how conjugates work. The new model suggests that carbohydrate presentation to T cells by antigen-presenting cells may strongly enhance the efficacy of antibody responses. Application of this principle in mouse models of group B streptococcal disease and Francisella tularensis infection by using a carrier peptide rather than a protein, resulted in vaccines more protective than standard glycoconjugate vaccines. We have also demonstrated the critical role of the peptide linker and the conjugation site in enhancing both the conjugation efficiency and the immune response to vaccines. The results support the notion that peptide glycoconjugate vaccines provide superior protection against bacterial challenges. Remarkably, the use of a peptide instead of a protein as a carrier confers better protection at much lower levels of carbohydrate specific IgG. This observation challenges the paradigm of a direct correlation between the amount of IgG induced by a glycoconjugate and protection. A comprehensive evaluation of antibodies and immune cells generated by protein vs peptide glycoconjugates will clarify the features that make peptide vaccines extremely potent. We will evaluate the antibody response to glycopeptide vaccines in terms of IgG subtypes, affinity, avidity and functional activity, as well as, elucidate the costimulatory/coinhibitory molecules being expressed on memory B cells and relate this to the functionality of the antibodies being made. This approach will define the characteristics that make glycopeptide-induced antibodies so potent and forms the basis for screening of B-cell hybridomas produced from peptide vs protein glycoconjugates. Correlates of highly functional antibody and critical B cell biomarkers will be identified. Hybridomas that produce antibodies with different degrees of protective activity will be evaluated by BCR sequencing. Fab fragments of low and highly effective antibodies will be co- crystallized with GBSIII oligosaccharide to investigate the role of epitope specificity in protection. We will apply our studies of these peptide/polysaccharide conjugates to create more efficacious and longer lasting immunity to glycoconjugate vaccines. ! !

阐明下一代糖缀合物的结构要求 项目摘要 糖缀合物疫苗在全球范围内提供巨大的健康益处，尽管它们的较少 在某些人群中成功的疾病风险很高。在大多数情况下，糖的结合 对载体蛋白的抗原已在经验上进行，对变量的关注很少 免疫反应。我们实验室的最新发现为如何结合提供了新的合理解释 工作。新模型表明，抗原呈递细胞向T细胞碳水化合物表现出来 强烈增强抗体反应的功效。该原理在B组的鼠标模型中的应用 链球菌疾病和francisella tularensis通过使用载体肽而不是蛋白质， 导致疫苗比标准糖缀合物疫苗更具保护性。我们也证明了 肽接头和结合位点的关键作用在提高共轭效率和 对疫苗的免疫反应。结果支持肽糖缀合物疫苗提供的观念 防止细菌挑战的优越保护。值得注意的是，使用肽而不是蛋白质作为一种 载体在碳水化合物特异性IgG的水平上赋予更好的保护。这个观察结果 挑战糖缀合物诱导的IgG量与 保护。蛋白质与肽产生的抗体和免疫细胞的全面评估 糖缀合物将阐明使肽疫苗极有效的特征。我们将评估 IgG亚型，亲和力，亲和力和功能活性对糖肽疫苗的抗体反应， 以及阐明在记忆B细胞上表达的共刺激/共抑制分子并相关 这是为了制作抗体的功能。这种方法将定义使得 糖肽诱导的抗体如此有效，并为筛查B细胞杂交瘤而形成基础 来自肽与蛋白质糖缀合物。高功能抗体和临界B细胞的相关性 将确定生物标志物。产生不同程度保护活性抗体的杂交瘤 将通过BCR测序评估。低和高效抗体的Fab片段将是共同的 用GBSIII寡糖结晶，以研究表位特异性在保护中的作用。我们将 应用我们对这些肽/多糖结合物的研究，以创造更有效，更持久的 对糖缀合疫苗的免疫力。呢 呢