Naturally Occurring Lipid Mediators Enhance Antibody Production

描述(由申请人提供)：一个新兴的概念是，炎症的消退是一个动态过程，对恢复体内平衡至关重要，从而预防慢性炎症和疾病。新近发现的由多不饱和脂肪酸(PUFA)衍生的脂质介质现在被认为是消炎的关键因素。这些内源性的专门化分解介体(SPM)组成了不同的家族，包括脂氧素、溶血素、保护素和乳脂素。SPM最近发现了调节免疫系统细胞的能力。体液反应对于保护机体免受微生物的侵袭是必不可少的，然而SPM在体液免疫中的作用及其对B细胞的影响尚未被研究过。这是一个重要的知识鸿沟。我们支持这一R21应用的新的试点数据表明，SPM具有增强丝裂原和抗原驱动的抗体反应的重要能力。这些结果支持我们的假设，即脂源性SPM通过促进B细胞功能来增强对感染的抗体应答，从而改善免疫记忆和长期保护。我们建议在感染和疫苗接种的临床前模型中，研究SPM对人和小鼠B细胞的免疫调节作用，以及在针对流感的适应性免疫反应中的作用。SPM的佐剂或“疫苗助推器”特性可以提高疫苗的效力和效用，允许在疫苗免疫原性较差或供应短缺时使用较小的剂量。我们提出了两个特定的目标，以开始揭示SPM对B细胞的功能。目的1：鉴定关键SPM刺激人B细胞抗体产生的能力。B细胞将被TLR和BCR配体以及选定的SPM激活。免疫球蛋白和免疫球蛋白抗体水平将与B细胞表型、存活和增殖分析一起测定。我们将研究接种季节性流感疫苗的受试者的B细胞记忆反应。这些研究将确定溶血素对记忆和抗体分泌B细胞群的影响。目的2：在临床前小鼠流感病毒疫苗接种和感染模型中，确定铅SPM刺激保护性抗体产生的能力。将使用小鼠流感疫苗接种和感染模型来分析SPM在适应性记忆反应中的功能。将使用三价季节性流感疫苗以及流感重组蛋白血凝素(HA)接种小鼠，并补充或不添加SPM。将测量免疫球蛋白和免疫球蛋白抗体滴度以及抗体的抑制和中和特性，以反映B细胞介导的适应性反应。将通过用活的流感病毒株(包括2009年H1N1流感病毒株)攻击免疫小鼠并监测体重减轻、存活率和病毒滴度来进一步评估SPM的刺激特性。

DESCRIPTION (provided by applicant): An emerging concept is that resolution of inflammation is a dynamic process crucial in restoring homeostasis, thus preventing chronic inflammation and disease. Newly identified lipid mediators, derived from polyunsaturated fatty acids (PUFA) are now recognized as crucial players in resolving inflammation. These endogenous specialized proresolution mediators (SPM) constitute separate families including lipoxins, resolvins, protectins and maresins. SPM have newly discovered abilities to regulate cells of the immune system. The humoral response is essential for protection against microorganisms, however the role of SPM in humoral immunity and their effect on B cells has not yet been studied. This is an important knowledge gap. Our new pilot data in support of this R21 application show that SPM have an important ability to enhance mitogen and antigen-driven antibody responses. These results support our hypothesis that the lipid-derived SPM enhance the antibody response to infection by promoting B cell function, leading to improved immune memory and long-term protection. We propose to study the immuno-regulatory functions of SPM on human and mouse B cells and during the adaptive immune response against influenza, in an infection and vaccination pre-clinical model. Adjuvant or "vaccine booster" properties of SPM could increase vaccine efficacy and utility, permitting smaller doses when a vaccine is poorly immunogenic or in short supply. We propose two specific aims to begin to reveal the function of SPM on B cells. Aim 1: Characterize the ability of key SPM to stimulate human B cell antibody production. B cells will be activated with TLR and BCR ligands along with selected SPM. IgM and IgG antibody levels will be determined along with B cell phenotype, survival and proliferation analyses. We will study B cell memory responses using seasonal influenza-vaccinated human subjects. These studies will determine the effects of resolvins on memory and antibody-secreting B cell populations. Aim 2: Determine the capacity of lead SPM to stimulate protective antibody production in a pre-clinical mouse influenza virus vaccination and infection model. The functions of SPM during an adaptive memory response will be analyzed using a mouse influenza vaccination and infection model. Mice will be vaccinated using the trivalent seasonal flu vaccine, as well as influenza recombinant protein, hemagglutinin (HA), and complemented with or without SPM. IgM and IgG antibody titers will be measured along with the antibodies' inhibitory and neutralizing properties, reflective of a B cell-mediated adaptive response. The stimulatory properties of SPM will be further assessed by challenging immunized mice with live influenza viral strains, including the 2009 H1N1, and monitoring weight loss, survival, and viral titers.