Development of a mucosal vaccine to prevent Clostridium difficile infection using papilloma pseudovirus as a vector.

使用乳头状瘤假病毒作为载体开发粘膜疫苗以预防艰难梭菌感染。

• 批准号: 10213890
• 负责人: Katherine L. Knight
• 金额: $ 50.23万
• 依托单位: LOYOLA UNIVERSITY CHICAGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10213890
• 关键词: Adjuvant; Affect; Amino Acid Sequence; Animal Disease Models; Antibiotic Therapy; Antibiotics; Antibodies; Antibody Formation; Antibody Response; Binding; C-terminal; Capsid; Cell surface; Cessation of life; Clinical; Clinical Trials; Clinical Trials Data Monitoring Committees; Clostridium difficile; DNA Viruses; Data; Development; Diarrhea; Disease; Environment; Epithelial; Epithelial Cells; Epithelium; FDA approved; Feces; Flagellin; Goals; Gold; Grant; Hamsters; Health Care Costs; Healthcare; Human; Human papillomavirus HPV L1 protein; Immunity; Immunize; Immunoglobulin A; Immunologic Memory; Incidence; Infection; Intestinal Mucosa; Intestines; L2 viral capsid protein; Life; Membrane Proteins; Memory; Mesocricetus auratus; Methods; Minor; Molecular Weight; Mucous Membrane; Mus; Nosocomial Infections; Oral; Papilloma; Papillomavirus; Pathogenesis; Patients; Phase; Phase II/III Clinical Trial; Phase II/III Trial; Plasmids; Probability; Proteins; Pseudomembranous Colitis; Recurrent disease; Reporting; Reproduction spores; Resistance; Surface; Surface Antigens; Symptoms; Targeted Toxins; Testing; Toxin; Vaccinated; Vaccination; Vaccines; Virulence Factors; Virus-like particle; alpha Toxin; anti-IgA; antitoxin; commensal microbes; disease transmission; efficacy testing; gut colonization; gut microbiota; health care settings; immunogenic; improved; in vivo; inhibiting antibody; mucosal vaccine; neutralizing antibody; particle; pathogen; prevent; programs; protein B; receptor binding; response; transmission process; vaccine development; vaccine efficacy; vector; vector vaccine

Abstract Clostridium difficile is a major cause of diarrhea in health care settings, with one-half million cases of infection, annually. Clostridium difficile infection and disease (CDI) usually occurs after antibiotic treatment, which eliminates much of the intestinal commensal microbiota and provides C. difficile the opportunity to colonize the gut. If the colonizing C. difficile produces toxins, many patients develop CDI and suffer from symptoms ranging from diarrhea to life-threatening pseudomembranous colitis. The incidence of CDI is on the rise, in large part because of increased use of antibiotics. Further, CDI recurs in ~20% of treated patients, making this a serious and expensive disease, for which there is no cure and no effective approved vaccine. The goal of this proposal is to develop a protective vaccine to prevent CDI and also, its transmission in the health care environment. While a few C. difficile vaccines are in Phase II/III clinical trials, they target the toxins, but will not prevent colonization of C. difficile in the intestinal mucosa, nor its transmission. We think that the optimal protective vaccine would be one that generates a strong mucosal IgA antibody response against toxins, and also one that prevents colonization of C. difficile. Papillomaviruses are small DNA viruses in which the major capsid protein, L1, assembles with the minor capsid protein, L2 into virus-like particles (VLP). These VLPs infect mucosal surfaces, are not infectious but are strongly immunogenic, making them ideal vectors for mucosal vaccines. We have developed papilloma pseudoviruses (PsV) from capsid proteins L1 and L2, that contain plasmids expressing the receptor binding domain (RBD) of the C. difficile toxins A and B, and we have preliminary data showing that these vaccines can induce neutralizing mucosal IgA and protect mice from challenge by C. difficile. In Aim 1, we will develop PsV expressing RBD of toxins A, B (from different strains) and binary toxin. We will immunize them in mice and hamsters. We will identify an adjuvant that will increase the mucosal Ab response and generate long-term immunologic memory in mice and hamsters, the gold standard for CDI animal models. In Aim 2, we will generate PsV expressing surface molecules FliD or high molecular weight SLP, molecules that will affect colonization, and determine if these vaccines can induce antibodies that inhibit colonization of C. difficile. In Aim 3, we will vaccinate mice and hamsters with a mixture of all the PsV and challenge with toxogenic C. difficile spores from different strains to determine if CDI and colonization of C. difficile are prevented. If successful, the mucosal vaccine will generate a robust neutralizing mucosal IgA response against both toxins and the surface antigens FliD and SLP. Such a vaccine would make a major contribution to improving the health care of thousands of patients in health care settings, and save billions of dollars in health care costs.

抽象的 艰难梭菌是医疗保健环境中腹泻的主要原因，有一半的感染病例， 每年。艰难梭菌感染和疾病（CDI）通常发生在抗生素治疗后，这 消除大部分肠道菌群，并为艰难梭菌提供了定殖的机会 肠道。如果艰难梭菌定植会产生毒素，许多患者会出现CDI并患有症状 从腹泻到威胁生命的假膜结肠炎。 CDI的发生率正在上升， 很大程度上是由于使用抗生素的使用增加。此外，CDI在约有20％的治疗患者中复发，这使得 一种严重且昂贵的疾病，无法治愈且没有有效批准的疫苗。目标的目标 提案是开发一种保护性疫苗，以防止CDI以及其在医疗保健中的传播 环境。虽然有几种艰难梭菌疫苗正在II/III期临床试验中，但它们靶向毒素，但会将 不能防止肠粘膜中艰难梭菌的定殖，也不能阻止其传播。我们认为最佳 保护性疫苗将是一种对毒素产生强粘膜IgA抗体反应的疫苗，也是一种 一种阻止艰难梭菌定植的一种。乳头瘤病毒是小的DNA病毒，其中主要是 衣壳蛋白L1用小的衣壳蛋白L2组装成病毒样颗粒（VLP）。这些VLP 感染粘膜表面，不是感染性的，但具有强烈的免疫原性，使其成为理想的媒介 粘膜疫苗。我们已经开发了来自衣壳蛋白L1和L2的乳头状病毒（PSV）， 包含表达艰难梭菌毒素A和B的受体结合结构域（RBD）的质粒，我们有 初步数据表明，这些疫苗可以诱导中和粘膜IgA并保护小鼠免受 艰难梭菌的挑战。在AIM 1中，我们将开发PSV表达毒素A，B的RBD（来自不同菌株） 和二进制毒素。我们将在老鼠和仓鼠中免疫它们。我们将确定一个将会增加的佐剂 粘膜AB反应并在小鼠和仓鼠中产生长期免疫记忆，黄金 CDI动物模型的标准。在AIM 2中，我们将产生表达表面分子或高高的PSV 分子量SLP，会影响定殖的分子，并确定这些疫苗是否可以诱导 抑制艰难梭菌定植的抗体。在AIM 3中，我们将用A接种老鼠和仓鼠 所有PSV的混合物和与来自不同菌株的疾病艰难梭菌孢子的挑战，以确定CDI是否是否 防止艰难梭菌的定殖。如果成功，粘膜疫苗将产生强大的 对毒素和表面抗原的粘膜IgA反应中和粘液反应。这样的疫苗 将为改善卫生保健环境中数千名患者的医疗保健做出重大贡献， 并节省数十亿美元的医疗保健费用。