A Lyme disease vaccine using newly identified in-vivo expressed antigens

使用新鉴定的体内表达抗原的莱姆病疫苗

• 批准号: 8601286
• 负责人: Michel Ledizet
• 金额: $ 29.57万
• 依托单位: L2 DIAGNOSTICS, LLC
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-01-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8601286
• 关键词: Acute; Adjuvant; Adverse effects; Animals; Antibiotic Therapy; Antibiotics; Antibodies; Antigens; Appearance; Arthritis; Arthropods; Bacteria; Bacterial Infections; Biochemical; Biological Assay; Bite; Black-legged Tick; Borrelia; Borrelia Infections; Borrelia burgdorferi; Carditis; Case Study; Centers for Disease Control and Prevention (U.S.); Control Groups; DNA; Development; Diagnosis; Diagnostic tests; Disease; Drug Formulations; Ensure; Enzyme-Linked Immunosorbent Assay; FDA approved; Gold; Human; Immune Sera; Immune response; Immunity; Immunization; Immunoblotting; Immunofluorescence Immunologic; Immunoglobulin G; Immunoglobulin M; Infection; LYMErix; Left; Life; Lyme Disease; Lyme Disease Vaccines; Measures; Membrane; Membrane Proteins; Methods; Mus; Neurologic; Order Spirochaetales; Organ; Patients; Phase; Population; Proteins; Protocols documentation; Public Health; Recombinant Proteins; Recombinants; Resolution; Sales; Sampling; Serologic tests; Skin; Staging; Symptoms; Syringes; System; Testing; Ticks; Time; TimeLine; Tissue Sample; Tissues; TiterMax; United States; Vaccines; Virulent; base; design; disorder control; in vivo; milligram; novel vaccines; prevent; public health relevance; research study; response; tissue culture; vaccine development

DESCRIPTION (provided by applicant): Lyme disease is the most common arthropod-borne disease in the United States, with over 38,000 confirmed and probable cases reported in 2009. The causative agent of Lyme disease is the spirochete Borrelia burgdorferi, which is transmitted by the tick of the Ixodes scapularis tick. The number of Lyme disease cases has tripled over the past fifteen years. The disease is sometimes difficult to diagnose in its early stages, leading to delayed treatment. Patients who are not treated in a timely fashion may suffer from severe, long lasting symptoms, and fail to respond to antibiotic therapy. Control measures designed to prevent contact with ticks are not sufficient to prevent the disease. We propose to develop a vaccine to control Lyme disease in human populations. After being injected into the skin of a mammalian host, Borrelia bacteria multiply locally before disseminating to other organs. We have now for the first time been able to isolate sufficient amounts of bacteria from the skin of infected mice in order to identify proteins expressed at high levels in the skin. We believe that immunization against these proteins, which are expressed in the early acute phase of Lyme disease, will be highly effective in preventing bacterial proliferation and dissemination. Fifteen membrane proteins identified in the skin of infected mice will be produced in recombinant form using standard bacterial expression systems. These proteins are conserved among strains of Borrelia, which should ensure that our vaccine is effective against all strains of circulating Borrelia. Each antigen will then be administered to C57Bl/6 mice in the presence of an adjuvant. We will demonstrate that this immunization protocol results in the appearance of anti-Borrelia antibodies. Immunized animals will then be challenged with 104 live Borrelia spirochetes. Twenty-one days after challenge we will collect tissue samples to determine whether they contain live Borrelia. We will also quantitatively detect the presence of Borrelia DNA in these samples. A protein antigen will be considered protective (1) if spirochetes cannot be cultured from the tissues of immunized mice after challenge or (2) if the bacterial load, determined from the amount of DNA detected, is reduced by at least 50%. At the conclusion of these experiments we will select up to four antigens for further study. Phase II experiments will determine the optimal formulation and method of delivery. We will also examine whether antigens can be combined to provide full protection against Borrelia.

描述（由申请人提供）：莱姆病是美国最常见的节肢动物传播疾病，2009 年报告了超过 38,000 例确诊和疑似病例。莱姆病的病原体是螺旋体伯氏疏螺旋体，由肩胛硬蜱传播。过去十五年来，莱姆病病例数量增加了两倍。这种疾病有时很难在早期阶段诊断出来，从而导致治疗延误。不及时治疗的患者可能会出现严重、长期持续的症状，并且对抗生素治疗无效。旨在防止接触蜱虫的控制措施不足以预防这种疾病。我们建议开发一种疫苗来控制人群中的莱姆病。注射到哺乳动物宿主的皮肤后，疏螺旋体细菌在传播到其他器官之前会在局部繁殖。我们现在第一次能够从受感染小鼠的皮肤中分离出足够量的细菌，以便识别皮肤中高水平表达的蛋白质。我们相信，针对这些在莱姆病早期急性期表达的蛋白质进行免疫接种，将非常有效地防止细菌增殖和传播。在受感染小鼠的皮肤中鉴定出的十五种膜蛋白将使用标准细菌表达系统以重组形式产生。这些蛋白质在伯氏疏螺旋体菌株之间是保守的，这应该确保我们的疫苗对所有流行的伯氏疏螺旋体菌株均有效。然后将每种抗原在佐剂存在下施用于C57Bl/6小鼠。我们将证明这种免疫方案会导致抗伯氏疏螺旋体抗体的出现。然后，免疫动物将接受 104 个活疏螺旋体螺旋体的攻击。攻击后二十一天，我们将收集组织样本以确定它们是否含有活的疏螺旋体。我们还将定量检测这些样本中是否存在疏螺旋体 DNA。 (1) 如果在攻击后不能从免疫小鼠的组织中培养出螺旋体，或者 (2) 如果根据检测到的 DNA 量确定的细菌负荷减少至少 50%，则蛋白质抗原将被认为具有保护性。在这些实验结束时，我们将选择最多四种抗原进行进一步研究。 II 期实验将确定最佳配方和给药方法。我们还将检查是否可以组合抗原以提供针对疏螺旋体的全面保护。