Immunotherapeutic Potential of Modified Lipooligosaccharides and Lipid A's

修饰脂寡糖和脂质 A 的免疫治疗潜力

基本信息

批准号：
8675799
负责人：
Robert K Ernst
金额：
$ 23.02万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-06-10 至 2016-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8675799
关键词：
Address Adjuvant Adjuvanticity Agonist Anabolism Animal Disease Models Animal Model Animals Antibody Formation Antigens Area Attenuated Baltimore Biological Assay Carbohydrates Cause of Death Cells Cervarix Chemicals Clinical Trials Complement Core Facility Data Development Disease Dose Drug Formulations Emulsions Engineering Enzymes Equilibrium Evaluation Excision FDA approved Fatty Acids Funding Future Goals Hepatitis B Virus Heterogeneity Human Human Papillomavirus Humoral Immunities Hydrolysis Immune response Immune system Immunity Immunologic Adjuvants Immunotherapeutic agent In Vitro Infection Infectious Agent Inflammation Influenza Intramuscular Investments Knowledge Lead Licensing Link Lipid A Lipids Location Malaria Maryland Mass Fragmentography Mass Spectrum Analysis Mediating Methodology Modeling Modification Mus Mutation Nature Oils Pharmacy facility Plague Preclinical Testing Process Production Property Reaction Reagent Regulator Genes Research Research Personnel Risk Salts Signal Transduction Specificity Structure T cell response Technology Testing Therapeutic Toxic effect Tuberculosis Tularemia Universities Vaccine Antigen Vaccine Design Vaccines Vertebral column Viral Viral Pathogenesis Virus Water Yersinia pestis aluminum sulfate base candidate selection cell type combat combinatorial chemistry cost effective design enzyme biosynthesis human TLR4 protein immunogenic in vivo inorganic phosphate interest lipooligosaccharide mimetics monophosphoryl lipid A novel novel strategies novel vaccines pathogenic bacteria preclinical study public health relevance research clinical testing response scaffold screening success toll-like receptor 4 vaccine development vaccine efficacy vaccinology

项目摘要

DESCRIPTION (provided by applicant): Protection from infectious agents known to be major causes of death worldwide, such as influenza, tuberculosis, and malaria, as well as potential release of bioweaponized agents which cause plague, tularemia, and meilloidosis require vaccines that generate humoral and T-cell responses. Effective component vaccines require the addition of adjuvants to increase their immunogenic capacities. Until recently alum salts, which require repeated applications and tend to be skewed towards T helper TH2-based immunity (humoral) rather than TH1, (cellular) were the only adjuvants approved for use in human vaccines. Recently, the lipid A mimetic (monophosphoryl lipid A, MPL) adjuvant has been combined with alum (AS04) in two FDA-approved vaccines (Cervarix (Human Papilloma Virus), and Fendrix (Hepatitis B Virus)). Additionally, synthetic lipid A mimetics aminoalkyl glucosaminide phosphates (AGPs) that also signal through Toll-Like Receptor 4 (TLR4) are being studied as both adjuvants or stand-alone immunogenic compounds. Thus, TLR4 agonists show great promise for use as adjuvants in component vaccines. However, the approved TLR4 agonist, MPL has distinct deficiencies both in potency and structural consistency and AGPs are labor intensive and costly to synthesize. Therefore, the goal of this revised R21 application is to undertake a novel approach of using Bacterial Enzymatic Combinatorial Chemistry (BECC) to make rationally-designed lipid A structures by modifying the lipid A structure of the lipooligosaccharide (LOS) from an attenuated (BSL-2 approved) Yersinia pestis (Yp) strain. This approach will use the non-stimulatory, hypoacylated, and bisphosphorylated lipid A structure present in LOS synthesized by this Yp strain as a scaffold to be modified by heterologous in trans expression of lipid A biosynthesis enzymes obtained from a wide variety of bacterial backgrounds with specificities for the removal or addition of fatty acid chain, phosphates moieties, and carbohydrates to the lipid A backbone, This process allow for the safe, cost effective, and efficient design of molecules with immunostimulatory potential. We will test the immunotherapeutic potential of these new molecules in vitro and in vivo to identify novel molecules representing adjuvants and/or immunomodulating reagents. We will also include well-characterized immunostimulants, such as MPL and known LPS structures as comparisons to the molecules synthesized by BECC. The successful demonstration of protective innate/adaptive immune responses by this novel approach of creating new adjuvants, in highly relevant in vivo animal model(s) could have important implications in the field of antigen recognition, formulation, and vaccine design.

描述（由申请人提供）：为了预防已知的全球主要死亡原因的传染性病原体，例如流感、结核病和疟疾，以及引起鼠疫、兔热病和类鼻疽病的生物武器制剂的潜在释放，需要能够产生体液免疫的疫苗。和 T 细胞反应。有效成分疫苗需要添加佐剂以提高其免疫原性能力。直到最近，明矾盐是唯一被批准用于人类疫苗的佐剂，因为明矾盐需要重复使用，并且往往偏向于基于辅助 T TH2 的免疫（体液）而不是 TH1（细胞）。最近，脂质 A 模拟物（单磷酰脂质 A，MPL）佐剂已与明矾 (AS04) 结合用于 FDA 批准的两种疫苗（Cervarix（人乳头瘤病毒）和 Fendrix（乙型肝炎病毒））。此外，也通过 Toll 样受体 4 (TLR4) 发出信号的合成脂质 A 模拟物氨烷基氨基葡萄糖胺磷酸盐 (AGP) 正在作为佐剂或独立的免疫原性化合物进行研究。因此，TLR4激动剂在成分疫苗中作为佐剂显示出巨大的应用前景。然而，已批准的 TLR4 激动剂 MPL 在效力和结构一致性方面都存在明显缺陷，并且 AGP 需要大量劳动力且合成成本高昂。因此，修订后的 R21 应用程序的目标是采用细菌酶组合化学 (BECC) 的新方法，通过修饰来自减毒（BSL-2 批准）鼠疫耶尔森氏菌 (Yp) 菌株的脂寡糖 (LOS) 的脂质 A 结构，来制造合理设计的脂质 A 结构。该方法将使用该 Yp 菌株合成的 LOS 中存在的非刺激性、低酰化和双磷酸化脂质 A 结构作为支架，通过从各种具有特异性的细菌背景中获得的脂质 A 生物合成酶的异源反式表达进行修饰用于在脂质 A 主链上去除或添加脂肪酸链、磷酸盐部分和碳水化合物，该过程可以安全、经济且高效地设计具有免疫刺激潜力的分子。我们将在体外和体内测试这些新分子的免疫治疗潜力，以鉴定代表佐剂和/或免疫调节试剂的新分子。我们还将包括充分表征的免疫刺激剂，例如 MPL 和已知的 LPS 结构，以与 BECC 合成的分子进行比较。通过这种创造新佐剂的新方法，在高度相关的体内动物模型中成功证明了保护性先天/适应性免疫反应，可能对抗原识别、制剂和疫苗设计领域产生重要影响。