The Role of Glycosylation in Regulating Immune Responses

糖基化在调节免疫反应中的作用

• 批准号: 8537477
• 负责人: THOMAS J TOLBERT
• 金额: $ 26.72万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-30 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8537477
• 关键词: Accounting; Amino Acid Sequence; Antibodies; Arthritis; Avidity; Binding; Biochemical; Biological Assay; Biological Models; Biotin; Carbohydrates; Cell surface; Cells; Cellular Assay; Chemicals; Complement; Complement-Dependent Cytotoxicity; Complex; Crystallization; Development; Disease; Engineering; Erythropoietin; Eukaryotic Cell; FDA approved; Family; Fc Receptor; Fluorescein; Folic Acid; Future; Genetic Engineering; Glycoproteins; Goals; Half-Life; Homologous Protein; Hormones; Human; Human Biology; IgG1; IgG2; IgG3; IgG4; Immune; Immune response; Immune system; Immunoconjugates; Immunoglobulin Fragments; Immunoglobulin G; In Vitro; Integrins; Knowledge; Label; Length; Ligand Binding; Ligands; Ligation; Link; Malignant Neoplasms; Measures; Mediating; Medical; Membrane; Methods; Modification; Peptides; Post-Translational Protein Processing; Prevalence; Production; Protein Glycosylation; Proteins; RGD (sequence); Relative (related person); Research; Role; Series; Serum Proteins; Site; Structure; Surface; Techniques; Technology; Testing; Yeasts; antibody-dependent cell cytotoxicity; antigen binding; base; biological systems; cancer cell; cellular targeting; cytotoxicity; folate-binding protein; glycosylated IgG; glycosylation; improved; in vivo; intercellular communication; milligram; novel; overexpression; protein function; public health relevance; receptor; receptor binding; research study; sialylation; small molecule; sugar; therapeutic development; therapeutic protein

DESCRIPTION (provided by applicant): The long term objectives of this project are to understand how changes in protein glycosylation regulate antibody dependent immune responses, and then to utilize that knowledge to develop improved antibody-based therapies for diseases such as cancer. The specific aims of this project are to: 1. Develop methods to produce homogeneous glycoproteins for the study and optimization of antibody glycosylation. 2. Determine the role of N-linked glycosylation in IgG Fc interactions with Fc receptors. 3. Develop methods to incorporate site-specific chemical modifications onto glycosylated IgG Fcs to facilitate biochemical studies and to modify bioactivity. 4. Determine the role of N-linked glycosylation in regulating antibody dependent cellular immune responses. As part of this research, novel methods will be developed to produce homogeneously glycosylated antibody fragments of the immunoglobulin G (IgG) subclasses, and also to attach small molecule receptor ligands to those antibody fragments site-selectively. The glycosylated and chemically modified antibody fragments thus produced will be utilized in in vitro biochemical and structural studies of antibody interactions with Fc receptors to determine the role of glycosylation and IgG subclass in regulating cellular immune responses. Antibody fragments attached to receptor ligands will be utilized to target cancer cells for antibody dependent immune responses in antibody dependent cell-mediated cytotoxicity (ADCC) assays and complement dependent cytotoxicity (CDC) assays. These cell-based assays will be used to optimize antibody dependent immune responses directed against target cells. In the future the knowledge gained by these experiments and the techniques developed will be valuable in developing antibody-based protein therapeutics directed against cancer and other diseases. PUBLIC HEALTH RELEVANCE: Antibodies are important therapeutic proteins that are used to treat cancers, arthritis, and many other diseases. The goal of this project is to understand how modification of human antibodies with sugars regulates antibody dependent immune responses, and then use the knowledge gained from these studies to develop better antibody-based treatments for diseases.

描述（由申请人提供）： 该项目的长期目标是了解蛋白质糖基化的变化如何调节抗体依赖性免疫反应，然后利用该知识来开发改进的基于抗体的抗体疗法（例如癌症）。该项目的具体目的是：1。开发产生均质糖蛋白的方法，以研究和优化抗体糖基化。 2。确定N连锁糖基化在IgG FC与FC受体相互作用中的作用。 3。开发将特异性化学修饰纳入糖基化IgG FC的方法，以促进生化研究并改变生物活性。 4。确定N连锁糖基化在调节抗体依赖性细胞免疫反应中的作用。作为这项研究的一部分，将开发出新的方法来产生免疫球蛋白G（IgG）亚类的均质糖基化抗体片段，并将小分子受体配体附着在这些抗体片段上。因此，因此产生的糖基化和化学修饰的抗体片段将用于抗体与FC受体相互作用的体外生化和结构研究，以确定糖基化和IgG亚类在调节细胞免疫反应中的作用。附着在受体配体上的抗体片段将用于靶向癌细胞，用于抗体依赖性免疫反应中，依赖性细胞介导的细胞介导的细胞毒性（ADCC）测定法和补体依赖性细胞毒性（CDC）测定法。这些基于细胞的测定将用于优化针对靶细胞的抗体依赖性免疫反应。将来，这些实验和开发的技术获得的知识对于开发针对癌症和其他疾病的基于抗体的蛋白质疗法将是有价值的。 公共卫生相关性： 抗体是重要的治疗蛋白，用于治疗癌症，关节炎和许多其他疾病。该项目的目的是了解用糖的人类抗体的修饰如何调节抗体依赖性免疫反应，然后利用从这些研究中获得的知识来开发更好的基于抗体的疾病治疗方法。

项目成果

Structural characterization of the Man5 glycoform of human IgG3 Fc.

• DOI: 10.1016/j.molimm.2017.10.001
• 发表时间: 2017-12
• 期刊: Molecular immunology
• 影响因子: 3.6
• 作者: Shah IS;Lovell S;Mehzabeen N;Battaile KP;Tolbert TJ
• 通讯作者: Tolbert TJ

Modular assembly of dimeric HIV fusion inhibitor peptides with enhanced antiviral potency.

• DOI: 10.1016/j.bmcl.2013.09.034
• 发表时间: 2013-11-15
• 期刊: BIOORGANIC & MEDICINAL CHEMISTRY LETTERS
• 影响因子: 2.7
• 作者: Xiao, Junpeng;Tolbert, Thomas J.
• 通讯作者: Tolbert, Thomas J.

Disulfide Bond Characterization of Endogenous IgG3 Monoclonal Antibodies Using LC-MS: An Investigation of IgG3 Disulfide-mediated Isoforms.

• DOI: 10.1039/c6ay01248e
• 发表时间: 2016-08-21
• 期刊: Analytical methods : advancing methods and applications
• 作者: Lakbub JC;Clark DF;Shah IS;Zhu Z;Go EP;Tolbert TJ;Desaire H
• 通讯作者: Desaire H

Conjugation site heterogeneity causes variable electrostatic properties in Fc conjugates.

• DOI: 10.1021/bc4000564
• 发表时间: 2013-06-19
• 期刊: BIOCONJUGATE CHEMISTRY
• 影响因子: 4.7
• 作者: Boylan, Nicholas J.;Zhou, Wen;Proos, Robert J.;Tolbert, Thomas J.;Wolfe, Janet L.;Laurence, Jennifer S.
• 通讯作者: Laurence, Jennifer S.

High-throughput biophysical analysis and data visualization of conformational stability of an IgG1 monoclonal antibody after deglycosylation.

• DOI: 10.1002/jps.23730
• 发表时间: 2013-11
• 期刊: JOURNAL OF PHARMACEUTICAL SCIENCES
• 影响因子: 3.8
• 作者: Alsenaidy, Mohammad A.;Kim, Jae Hyun;Majumdar, Ranajoy;Weis, David D.;Joshi, Sangeeta B.;Tolbert, Thomas J.;Middaugh, C. Russell;Volkin, David B.
• 通讯作者: Volkin, David B.