Immune Enhancing Echinacea Bacterial Endophytes

描述(申请人提供)：紫锥菊是美国草药市场上使用最广泛的免疫刺激植物之一。然而，临床疗效评估遇到了不一致的结果，很难解释，因为他们使用了 种类繁多的化学成分不明确的制剂。此外，尽管进行了数十年的研究，但尚不清楚紫锥菊对感冒和流感的主要治疗作用是免疫刺激、抗炎还是两者兼而有之。动物研究表明，这种植物具有治疗作用，紫锥菊烷基酰胺是主要的抗炎成分。紫锥菊多糖曾被认为是这种植物的免疫增强作用，但目前存在于如此低的水平，并且在体外表现出极弱的天然免疫激活潜力，因此不太可能具有治疗作用。我们发现，紫锥菊和其他免疫增强型植物体内的细菌成分主要是脂多糖和布劳恩型脂蛋白，它们是体内和体外天然免疫细胞激活剂，而内生细菌是这些成分的主要来源。这两种细菌成分水平的差异是从北美六个主要种植者/商业供应商获得的紫锥菊提取物在天然免疫细胞激活方面观察到的实质性差异(~100倍)的原因。细菌成分含量/活性的这种巨大变异性可能是一个促成因素 对于紫锥菊临床试验中表现出的不一致的结果，因为使用口服细菌制剂(如益生菌)的研究已经证明了与免疫相关的益处。我们的假设是紫锥菊中的内生细菌是脂多糖和布朗类脂蛋白的主要来源，这些内生细菌的数量/类型的差异是不同紫锥菊植物材料免疫增强潜力差异的原因。我们的初步研究已经鉴定出23种细菌内生植物，这些菌株的提取物的能力差异很大(高达10,000倍) 在体外激活巨噬细胞。目标1将确定紫锥菊中的哪些细菌内生菌是植物材料所展示的活性的主要贡献者。目的2通过口服内生细菌的数量/种类与该植物材料中发现的相同数量/类型的内生细菌，来确定紫锥菊对小鼠几种免疫参数的体外活性和体内作用是否可以被模仿。目标3将确定影响紫花锦鸡儿的农艺因素 植物活性，并将定义内生菌含量/活性的变化，负责活动变化。实现这些目标将使人们更好地了解紫锥菊如何影响先天免疫系统，并通过提供信息和工具来更恰当地标准化紫锥菊产品以用于临床研究，从而影响CAM研究。这项研究还将增加越来越多的证据，表明口服某些类型的细菌或细菌成分可以对人类健康产生重大影响。 与公共健康相关：紫锥菊是美国使用最广泛的植物补充剂之一。这项拟议的研究将确定这种植物中的内生细菌，这些细菌是负责增强天然免疫功能的细菌成分的主要来源。这项研究还将增加越来越多的证据，表明口服某些类型的细菌或细菌成分可以对人类健康产生重大影响。

DESCRIPTION (provided by applicant): Echinacea species represent one of the most widely used immunostimulatory botanicals in the US herbal market. However, clinical evaluation of efficacy has met with inconsistent results that are difficult to interpret because they have used a wide variety of chemically ill-defined preparations. In addition, despite decades of research, it i not known if Echinacea's main therapeutic action on colds and flu is immunostimulatory, anti-inflammatory or a combination of both. Animal research indicates that this botanical exerts both therapeutic actions and Echinacea alkylamides represent the major anti-inflammatory component. Echinacea polysaccharides, once thought to be responsible for the immune enhancing action of this plant, are present at such low levels and exhibit extremely weak in vitro innate immune activation potential and thus are not likely to be therapeutically relevant. We have found that the bacterial components lipopolysaccharides and Braun-type lipoproteins are major in vitro and in vivo innate immune cell activators within Echinacea and other immune enhancing botanicals and those endophytic bacteria represent the principal source of these components. Differences in the levels of these two bacterial components are responsible for the substantial variation observed (~100-fold) in innate immune cell activation from E. purpurea extracts of material obtained from six major growers/commercial suppliers in North America. This substantial variability in bacterial component content/activity could be a factor contributing to the inconsistent outcomes exhibited in Echinacea clinical trials since immune related benefits have been demonstrated in studies employing orally administered bacterial preparations such as probiotics. Our hypothesis is that endophytic bacteria within E. purpurea are a major source of LPS and Brawn type lipoproteins and variation in the amount/type of these endophytes are responsible for differences in the immune enhancing potential of diverse Echinacea plant material. Our preliminary studies have identified 23 bacterial endophytic species within E. purpurea and extracts from these isolates varied substantially (up to 10,000-fold) in their ability to activate macrophages in vitro. Aim 1 will identify which bacterial endophytes within E. purpurea are principal contributors to the activity exhibited by the plant material. Aim 2 will determine if the in vitro activity and in vivo action of E. purpurea on several immune parameters in mice can be mimicked by the oral ingestion of endophytic bacteria equivalent in amount/type to that found in this plant material. Aim 3 will identify agronomic factors influencing E. purpurea plant activity and will define shifts in endophyte content/activity responsible for activity change. Accomplishing these objectives will provide a greater understanding of how Echinacea influences the innate immune system and will impact CAM research by providing the information and tools to more appropriately standardize Echinacea products for use in clinical studies. This research will also add to the growing body of evidence that indicates that the oral consumption of certain types of bacteria or bacterial components can have a major impact on human health. PUBLIC HEALTH RELEVANCE: Echinacea is one of the most widely used botanical supplements in the US. The research proposed will identify endophytic bacteria within this botanical that are the main source of bacterial components responsibe for enhancing innate immune function. This research will also add to the growing body of evidence that indicates that the oral consumption of certain types of bacteria or bacterial components can have a major impact on human health.