Entirely Carbohydrate Vaccine Constructs and Their Application in Probing Glycoim

全碳水化合物疫苗构建体及其在检测糖蛋白中的应用

• 批准号: 8730098
• 负责人: Peter R Andreana
• 金额: $ 29.29万
• 依托单位: UNIVERSITY OF TOLEDO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8730098
• 关键词: Affinity; Anaerobic Bacteria; Antibodies; Antibody Specificity; Antigens; Area; B-Lymphocytes; Bacterial Polysaccharides; Binding; Biological Assay; Breast Cancer Cell; Breast Carcinoma; C57BL/6 Mouse; CD4 Positive T Lymphocytes; Cancer Vaccines; Cancer cell line; Carbohydrate Chemistry; Carbohydrates; Carrier Proteins; Cell surface; Cells; Cellular Immunity; Charge; Chemicals; Chemistry; Complement; Complement-Dependent Cytotoxicity; Complex; Conjugate Vaccines; Development; Enzyme-Linked Immunosorbent Assay; Evaluation; Event; Flow Cytometry; Glycoproteins; Goals; Histocompatibility Antigens Class II; Human; Humoral Immunities; Hydrazones; Immune; Immune response; Immune system; Immunity; Immunization; Immunoglobulin G; Immunoglobulin M; In Vitro; Knowledge; Lactate Dehydrogenase; Lead; Lipids; MCF7 cell; Malignant Neoplasms; Mediating; Medicine; Methodology; Methods; Modeling; Modification; Monosaccharides; Mus; Neoplasm Metastasis; Oximes; Pathway interactions; Peptides; Polysaccharides; Prevention therapy; Preventive; Process; Property; Proteins; Research; Role; Route; Scheme; Serum; Site; Solid Neoplasm; Specificity; Structure; Subcutaneous Injections; Survival Rate; T-Lymphocyte; Testing; Therapeutic; Tumor Cell Line; Tumor-Associated Carbohydrate Antigens; U251; Vaccine Research; Vaccines; Work; arm; base; cancer cell; cancer therapy; cell killing; cytotoxicity; design; disorder prevention; efficacy testing; immune function; immunogenic; immunogenicity; improved; in vivo; innovation; insight; killings; novel; oxidation; programs; research study; response; sugar; tool; tumor; tumor growth; vaccine candidate; vaccine development; Affinity; Anaerobic Bacteria; Antibodies; Antibody Specificity; Antigens; Area; B-Lymphocytes; Bacterial Polysaccharides; Binding; Biological Assay; Breast Cancer Cell; Breast Carcinoma; C57BL/6 Mouse; CD4 Positive T Lymphocytes; Cancer Vaccines; Cancer cell line; Carbohydrate Chemistry; Carbohydrates; Carrier Proteins; Cell surface; Cells; Cellular Immunity; Charge; Chemicals; Chemistry; Complement; Complement-Dependent Cytotoxicity; Complex; Conjugate Vaccines; Development; Enzyme-Linked Immunosorbent Assay; Evaluation; Event; Flow Cytometry; Glycoproteins; Goals; Histocompatibility Antigens Class II; Human; Humoral Immunities; Hydrazones; Immune; Immune response; Immune system; Immunity; Immunization; Immunoglobulin G; Immunoglobulin M; In Vitro; Knowledge; Lactate Dehydrogenase; Lead; Lipids; MCF7 cell; Malignant Neoplasms; Mediating; Medicine; Methodology; Methods; Modeling; Modification; Monosaccharides; Mus; Neoplasm Metastasis; Oximes; Pathway interactions; Peptides; Polysaccharides; Prevention therapy; Preventive; Process; Property; Proteins; Research; Role; Route; Scheme; Serum; Site; Solid Neoplasm; Specificity; Structure; Subcutaneous Injections; Survival Rate; T-Lymphocyte; Testing; Therapeutic; Tumor Cell Line; Tumor-Associated Carbohydrate Antigens; U251; Vaccine Research; Vaccines; Work; arm; base; cancer cell; cancer therapy; cell killing; cytotoxicity; design; disorder prevention; efficacy testing; immune function; immunogenic; immunogenicity; improved; in vivo; innovation; insight; killings; novel; oxidation; programs; research study; response; sugar; tool; tumor; tumor growth; vaccine candidate; vaccine development

DESCRIPTION (provided by applicant): The principal goal of this research program is to develop practical synthetic approaches for the modification of a unique capsular zwitterionic polysaccharide (ZPS), PS A1, and conjugate tumor-associated carbohydrate antigens (TACAs) to develop entirely carbohydrate vaccine constructs. PS A1 is a structurally well- characterized polysaccharide containing a repeating tetrasaccharide core isolated from the capsular polysaccharide complex (CPC) of the anaerobe Bacterioides fragilis NCTC 9343. It was determined that PS A1 could coax an immune system to elicit cellular and humoral immune functions via T- and B-cell involvement that influenced helper and memorial response events - a paradigm shifting discovery. What is not entirely clear is the immunogenicity of entirely carbohydrate-based constructs as an alternative approach to glycoprotein conjugates for tumor vaccine development. This research program, therefore, aims to study and assess the innovative vaccine constructs using three specific aims: 1) To develop efficient conjugation methods for the construction of structurally well-defined TACA-PS A1 conjugate vaccines, 2) To study the immunological properties of TACA-PS A1 conjugates in mice and evaluate the specificity and cytotoxicity of TACA-PS A1- raised Abs to human cancer cell lines, and 3) To evaluate, in vivo, the efficacy of TACA-PS A1-invoked immunity to treat tumors. Anti-sera, obtained from murine immunizations will be used to determine total antibody (Ab) titers and Ab isotypes by ELISA, relaying information about immunity. Furthermore, ELISA will be used to determine Ab specificity against conjugated TACAs. Flow cytometry will be used to validate Ab specificity with human cancer cell lines. Complement-mediated cell killing (cytotoxicity) analysis using human tumor cell lines, will reveal Ab function. Finally, assessing and evaluating the efficacy of select TACA-PS A1 constructs to treat cancer will be demonstrated through in vivo experiments utilizing murine models grafted with the TA3/Ha murine breast carcinoma. The broader impact of the proposed research aims to demonstrate how entirely carbohydrate-based vaccines elicit cellular and humoral responses in the immune system. This research program will contribute to improved methodologies for the field of glycoconjugation chemistry, a better understanding of the role of PS A1 as an immunogen in eliciting immune responses, and assessment and evaluation of cancer specific constructs that are rationally designed. This project will, therefore, have an important impact on cancer treatment by providing a novel carbohydrate-based agent for preventive and therapeutic medicine and increase our understanding of glycoimmunology. We also expect this program to be broadly applicable to other areas of vaccine research.

描述（由申请人提供）：该研究计划的主要目标是开发实用的合成方法，以修饰独特的囊膜Zwitterionic多糖（ZPS），PS A1和共轭肿瘤相关的碳水化合物抗原（TACAS），以开发完全碳水化合物疫苗的构造。 PS A1 is a structurally well- characterized polysaccharide containing a repeating tetrasaccharide core isolated from the capsular polysaccharide complex (CPC) of the anaerobe Bacterioides fragilis NCTC 9343. It was determined that PS A1 could coax an immune system to elicit cellular and humoral immune functions via T- and B-cell involvement that influenced helper and memorial response events - a范式转移发现。尚不完全清楚的是，完全基于碳水化合物的构建体的免疫原性是用于糖蛋白结合物用于肿瘤疫苗发育的替代方法。因此，该研究计划旨在使用三个特定目的研究和评估创新的疫苗构建：1）开发有效的结合方法，用于构建结构定义明确的TACA-PS A1 A1 Conjugate疫苗，2）研究小鼠taca-ps a1的免疫学特性，并评估人类的特异性和cy ox cy to cy of to to and cy of cy of time cy of timity and cya cy of cytox cytox cytox cytox cytox cytox cytox cytox cytox cytox，细胞系和3）在体内评估TACA-PS A1-IN-INVOKENS免疫治疗肿瘤的功效。通过鼠免疫获得的抗sera将用于确定ELISA的总抗体（AB）滴度和AB同型，并传达有关免疫力的信息。此外，ELISA将用于确定针对共轭炸玉米饼的特异性。流式细胞仪将用于通过人类癌细胞系来验证特异性。使用人肿瘤细胞系补体介导的细胞杀伤（细胞毒性）分析将揭示AB功能。最后，将通过使用TA3/HA鼠乳腺癌接枝的鼠模型进行体内实验来证明和评估精选TACA-PS A1构建体治疗癌症的疗效。拟议研究的更广泛的影响旨在证明如何完全基于碳水化合物的疫苗在免疫系统中引起细胞和体液反应。该研究计划将有助于改进糖结化学领域的方法学，更好地理解PS A1作为免疫反应中免疫反应的作用，以及对理性设计的癌症特定结构的评估和评估。因此，该项目将通过提供一种新型的基于碳水化合物的药物来预防和治疗医学，并增强我们对糖免疫学的理解，从而对癌症治疗产生重要影响。我们还希望该计划广泛适用于疫苗研究的其他领域。