Humoral and adaptive immune responses study in Vibrio cholerae infection

霍乱弧菌感染的体液和适应性免疫反应研究

• 批准号: 9189121
• 负责人: Md Taufiqur Rahman Bhuiyan
• 金额: $ 7.17万
• 依托单位: INTERNATIONAL CTR/DIARRHOEAL DIS RES
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-22 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9189121
• 关键词: Address; Affect; Affinity; Agar; Anti-Bacterial Agents; Antibodies; Antibody Response; Antigens; Area; B-Lymphocytes; Bangladesh; Belief; Biological Assay; Biopsy Specimen; Blood specimen; Boston; Cells; Child; Cholera; Cholera Toxin; Cholera Vaccine; Cloning; Coculture Techniques; Collaborations; Development; Disease; Educational process of instructing; Flagella; Flow Cytometry; General Hospitals; Genetic Variation; Genomics; Goals; Grant; Heating; Helper-Inducer T-Lymphocyte; Human; Immune; Immune response; Immunity; Immunoglobulin A; Immunoglobulin Class Switching; Immunoglobulin G; Immunoglobulin M; Immunoglobulin Somatic Hypermutation; Immunoglobulin Switch Recombination; Immunoglobulins; Immunologic Techniques; Immunology; Individual; Infection; Institutes; International; Knowledge; Liquid substance; Lymphocyte; Manuscripts; Massachusetts; Measures; Mediating; Mediator of activation protein; Memory; Memory B-Lymphocyte; Mentorship; Microbiology; Microscopy; Molecular Biology; Mononuclear; Morbidity - disease rate; Mutation; O Antigens; Onset of illness; Oral; Pathway interactions; Patients; Plasma Cells; Plasmablast; Predisposition; Preparation; Production; Property; Proteins; Proteomics; Research; Research Personnel; Research Training; Resources; Serum; Source; Surface; T-Lymphocyte; Techniques; Time; Toxin; Training; Training Programs; Universities; Vaccination; Vibrio cholerae; Western Blotting; Work; adaptive immunity; career development; cell motility; data management; experience; falls; graduate student; in vivo; international center; mortality; nano; novel; peripheral blood; programs; response; single cell analysis; skills

PROJECT SUMMARY Cholera is a severe dehydrating disease caused by Vibrio cholerae. Unfortunately, we do not yet know what is the prime functional mediator of protection against cholera. This deficit has impacted development of cholera vaccines, with currently available cholera vaccines providing lower-level and shorter-term protection against cholera, especially in children, than that afforded by previous wild type disease. We and others have generated strong preliminary evidence that protective immunity against cholera is mediated by antibodies at the mucosal surface, and that V. cholerae O-specific polysaccharide (OSP) is the primary antigenic target. V. cholerae is a motile non-invasive enteropathogen with a single polar sheathed flagellum (covered by LPS/OSP). We hypothesize that differences in the follicular T cell immune response between vaccinees and infected cholera patients suppress the immune responses in vaccinees. In addition, we also hypothesize that these differences affect the development, maturation, and functionality of antibody responses against key Vibrio cholerae antigens, O-specific polysaccharide (OSP) and cholera toxin (CT). To address these hypotheses, we propose three specific aims. Aim #1: We will use flow cytometry of mononuclear cells isolated from patients with cholera to evaluate follicular T cells (TFH) in relation to infection at different days after onset of disease and following vaccination. Aim #2: We will use a novel nano- well approach to isolate single cholera antigen-specific cells by phenotypic characterization in duodenal biopsy specimens (and peripheral blood) of individuals recovering from cholera. We will clone the immunoglobulin chains from these recovered lymphocytes, including plasmablasts, plasma cells, and memory B cells, and we will evaluate cloned antibodies for functionality, as well as affinity maturation and somatic hyper-mutation; we will perform a similar analysis of blood samples of vaccinees. Aim #3: We will assess for the presence of V. cholerae agglutinating and anti-motility antibodies in patients with cholera, as well as in vaccinees. This work builds upon a highly-successful and ongoing international collaborative effort between researchers at the International Centre for Diarrhoeal Disease Research in Dhaka, Bangladesh (icddr,b), the Massachusetts General Hospital- Harvard in Boston and Emory University, Atlanta, USA. The goal of this K43 Emerging Global Leader Career Development Program would be to strengthen the skills necessary to become an independent researcher/investigator in mucosal immunology in resource-limited settings.

项目摘要 霍乱是一种由弧菌霍乱引起的严重脱水疾病。不幸的是，我们还没有 知道什么是针对霍乱的主要功能介体。这种赤字影响了 霍乱疫苗的开发，目前可用的霍乱疫苗可提供较低层次的疫苗 对霍乱，尤其是在儿童中，比以前的野生类型所提供的短期保护尤其是在儿童中 疾病。我们和其他人产生了强有力的初步证据，保护性免疫力 霍乱是由粘膜表面的抗体介导的，而V. cholerae o特异性 多糖（OSP）是主要的抗原靶标。 V. Cholerae是一种流动性无创的 肠病，带有单个极性鞘鞭毛（由LPS/OSP覆盖）。我们假设这一点 疫苗和感染霍乱患者之间卵泡T细胞免疫反应的差异 抑制疫苗的免疫反应。此外，我们还假设这些 差异会影响抗体反应对密钥的发展，成熟和功能 Vibrio霍乱抗原，O特异性多糖（OSP）和霍乱毒素（CT）。解决这些 假设，我们提出了三个具体目标。目标＃1：我们将使用单核的流式细胞仪 从霍乱患者中分离出的细胞，以评估与感染有关的卵泡T细胞（TFH） 疾病发作和疫苗接种后的不同日子。 AIM＃2：我们将使用一种新颖的纳米 - 通过在 从霍乱中恢复的个体的十二指肠活检标本（和外周血）。我们将 克隆这些回收的淋巴细胞中的免疫球蛋白链，包括血浆，血浆 细胞和记忆B细胞，我们将评估克隆的抗体功能以及亲和力 成熟和体细胞超名；我们将对血液样本进行类似的分析 疫苗。目的＃3：我们将评估霍乱弧菌的存在和抗动力 霍乱和疫苗患者的抗体。这项工作以高度成功的为基础 以及国际国际中心研究人员之间正在进行的国际合作努力 孟加拉国达卡（ICDDR，b）的腹泻病研究，马萨诸塞州综合医院 - 哈佛在美国亚特兰大的波士顿和埃默里大学。这个K43新兴全球领导者的目标 职业发展计划是增强成为独立的技能 资源有限设置中的粘膜免疫学研究人员/研究者。