Activation of semi-invariant and diverse NKT cells with an adjuvant combination

使用佐剂组合激活半不变且多样化的 NKT 细胞

• 批准号: 10053313
• 负责人: Mark L Lang
• 金额: $ 36.71万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-11-06 至 2022-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10053313
• 关键词: Adjuvant; Affect; Alhydrogel; Antibody-mediated protection; Antigen Presentation; Antigens; Attenuated; Autoimmunity; B-Lymphocytes; Bacterial Toxins; Binding; Cell surface; Cells; Cellular Immunity; Clostridium difficile; Communities; Complex; Cytotoxic T-Lymphocytes; Equilibrium; FDA approved; Family; Formulation; Glycolipids; Haptens; Health; Human; Humoral Immunities; Immune response; Immune system; Immunity; Immunization; Immunize; In Vitro; Infection prevention; Inflammasome; Inflammatory; Keyhole Limpet Hemocyanin; Knowledge; Life; Ligands; Link; Lipids; Longevity; Malignant Neoplasms; Measurement; Measures; Memory B-Lymphocyte; Mission; Modeling; Modification; Molecular; Monitor; Mus; Natural Immunity; Organ; Patients; Pattern; Peptides; Peripheral Blood Mononuclear Cell; Phase I Clinical Trials; Phosphotransferases; Plasma Cells; Production; Proteins; Public Health; Reporting; Research; Safety; Series; Signal Transduction; Structure; T-Cell Activation; Testing; Tetanus Toxoid; Toxic effect; Toxin; United States National Institutes of Health; Vaccination; Vaccine Adjuvant; Vaccine Antigen; Vaccines; Work; adaptive immune response; adaptive immunity; alpha-galactosylceramide; aluminum sulfate; cytokine; experimental study; in vivo; innovation; knowledge base; pathogen; recruit; response; single cell analysis; transcription factor; vaccine candidate

ABSTRACT There are several safe candidate adjuvants that potentiate beneficial immunity following vaccination. However, single adjuvant formulations have performed quite modestly in terms of stimulating the desired blend of humoral versus cellular immunity and/or Th1 versus Th2 immunity. Combination adjuvants offer a possible way forward, but the mechanisms of action of such combinations remain poorly defined. Therefore, we will combine the FDA-approved adjuvant Alum (alhydrogel) with the CD1d-binding glycolipid adjuvant α-galactosylceramide (α-GC) and delineate the mechanisms of action with a particular emphasis on CD1d-restricted Natural Killer T (NKT) cells and humoral immunity. Alum is safe and stimulates excellent Th2 but poor Th1 immunity. We discovered that the Th2 response to Alum depends in large part upon diverse TCR-expressing Type II NKT cells (dNKT) which recognize CD1d/glycolipid complexes but do not recognize the α-GC adjuvant. The α-GC adjuvant stimulates semi-invariant TCR-expressing Type I NKT cells (iNKT) and results in a mixed Th1/Th2 response. It is already known from Phase I clinical trials in patients with cancer that α-GC is well-tolerated and safe. Furthermore, modification of the α-GC structure can readily be employed to skew the Type I NKT- dependent Th1/Th2 balance. Our objective for this proposal is to test the central hypothesis that the combination of Alum and α-GC leads to CD1d/glycolipid presentation and a coordinated dNKT and iNKT- driven mixed Th1/Th2 response against vaccine antigens. In Specific Aim 1, we will perform a series of in vitro studies in primary murine and human cells to determine how the adjuvant combination affects CD1d Ag presentation and activation and functional differentiation of NKT cells into different effector subsets. We will also examine the effect of adjuvant combination on class I and class II presentation of co-administered protein antigens. In Specific Aim 2, we will undertake a series of in vivo experiments in mice to examine the functional consequences of adjuvant combination for Ab-mediated protection against bacterial toxins. Experiments to analyze cellular as well as humoral immunity will be included as the project develops and in vivo toxicity (or lack thereof) will be determined by measurement of pro-inflammatory cytokines as well as markers of autoimmunity and organ damage. We feel this project is innovative because it will provide the vaccine community with mechanistic information when CD1d-binding adjuvants are combined with Alum adjuvant and give careful consideration to NKT cell- driven components of the immune response. We feel the project is significant because it will illuminate potential avenues for inclusion of CD1d-binding glycolipids in mixed adjuvant platforms.

抽象的 有几种安全的候选佐剂可以增强疫苗接种后的有益免疫力。 单一佐剂制剂在刺激所需的混合剂方面表现相当有限 体液免疫与细胞免疫和/或 Th1 与 Th2 免疫的组合佐剂提供了一种可能的方法。 向前发展，但这种组合的作用机制仍然不明确，因此，我们将结合起来。 FDA 批准的佐剂明矾（水凝胶）与 CD1d 结合糖脂佐剂 α-半乳糖神经酰胺 (α-GC) 并描述作用机制，特别强调 CD1d 限制性自然杀伤 T (NKT) 细胞和体液免疫是安全的，可刺激出色的 Th2 免疫，但可刺激较差的 Th1 免疫。 发现 Th2 对 Alum 的反应在很大程度上取决于表达不同 TCR 的 II 型 NKT 识别 CD1d/糖脂复合物但不识别 α-GC 佐剂的细胞 (dNKT)。 佐剂刺激表达半不变 TCR 的 I 型 NKT 细胞 (iNKT) 并产生混合的 Th1/Th2 从癌症患者的 I 期临床试验中已经知道，α-GC 具有良好的耐受性和疗效。 此外，α-GC 结构的修饰可以很容易地用于扭曲 I 型 NKT-。 我们此提案的目标是检验中心假设： 明矾和 α-GC 的组合导致 CD1d/糖脂呈递以及协调的 dNKT 和 iNKT- 驱动针对疫苗抗原的混合 Th1/Th2 反应。 在具体目标 1 中，我们将在原代小鼠和人类细胞中进行一系列体外研究，以确定 佐剂组合如何影响 CD1d Ag 的呈递、激活和功能分化 我们还将研究佐剂组合对 I 类和 NKT 细胞的影响。 共同施用的蛋白质抗原的 II 类呈递。 在具体目标2中，我们将在小鼠身上进行一系列的体内实验来检查功能 佐剂组合对抗体介导的细菌毒素保护实验的影响。 随着项目的发展，将包括分析细胞和体液免疫以及体内毒性（或 缺乏）将通过测量促炎细胞因子以及标记物来确定 自身免疫和器官损伤。 我们认为这个项目是创新的，因为它将为疫苗界提供机械信息 当CD1d结合佐剂与明矾佐剂组合并仔细考虑NKT细胞时 我们认为该项目很重要，因为它将阐明免疫反应的驱动组成部分。 将 CD1d 结合糖脂纳入混合佐剂平台的潜在途径。