Campylobacter jejuni outer membrane protein vaccine

描述(由申请人提供)：空肠弯曲杆菌是美国细菌性胃肠炎的主要原因，由于其能够引起食源性和水源性疫情，已被美国国立卫生研究院列为B类生物恐怖分子。美国每年至少有240万例空肠弯曲菌病，发病率超过沙门氏菌和志贺氏菌的总和(5)。空肠弯曲菌感染也是Guiilain-Barre综合征(GBS)最常见的前驱事件，GBS是一种明显由针对空肠弯曲菌表面抗原的自身免疫反应引起的急性运动瘫痪。因此，非常需要一种有效的空肠弯曲菌疫苗，以保护美国民众免受自然发生的空肠弯曲菌疾病和潜在的生物恐怖袭击的影响。基于空肠弯曲菌全细胞制剂的疫苗已经被提出，然而，由于关于GBS发展的不确定性，有必要采用替代方法。因此，由高度保守的外膜蛋白(OMP)组成的疫苗可能最有希望安全地诱导保护性免疫，而不会诱发GBS。 众所周知，空肠弯曲菌具有高度的变异性，这肯定会影响蛋白质亚单位疫苗的开发。一种适合于疫苗纳入的蛋白质必须在尽可能大比例的空肠弯曲菌菌株中保存，必须具有免疫原性，并且必须诱导对大量不同的空肠弯曲菌菌株的保护性免疫。虽然对空肠弯曲菌基因组序列的分析有助于了解其OMPS的互补，但只有通过蛋白质组分析直接鉴定OMPS才能提供有关空肠弯曲菌菌株实际表达的OMPS的详细信息。 假设：某些空肠弯曲菌外膜蛋白(OMP)在所有空肠弯曲菌株中都是保守的，在人类感染期间具有免疫原性，并将产生保护性免疫反应。 具体目的1.利用蛋白质组学和质谱法对几株空肠弯曲菌外膜的蛋白质组分进行鉴定。 具体目的2.我们将确定在我们最初的菌株中保守的OMP是否也在大量的空肠弯曲菌菌株(新鲜的临床分离株和美国各地的档案菌株收集)中发现，并将评估感染的人对这些OMP的免疫反应。 具体目的3.我们将确定用保守的、纯化的重组OMPS免疫小鼠是否能保护随后的实验性空肠弯曲菌感染。

DESCRIPTION (provided by applicant): Campylobacter jejuni is the leading cause of bacterial gastroenteritis in the U.S., and has been classified by the NIH as a Category B Bioterrorism Agent due to its ability to cause food-borne and water-borne outbreaks. There are at least 2.4 million cases of C. jejuni disease in the U.S. annually, with an incidence exceeding that of Salmonella and Shigella combined (5). C. jejuni infection is also the most common antecedent event to the development of Guiilain-Barre Syndrome (GBS), an acute motor paralysis that apparently results from an autoimmune response directed against C. jejuni surface antigens. An effective vaccine against C. jejuni is therefore highly desirable, to protect the U.S. population from both naturally-occurring C. jejuni disease and that arising from potential bioterrorist attacks. Vaccines based on C. jejuni whole-cell preparations have been proposed, however, due to uncertainties concerning the development of GBS, alternative approaches are warranted. A vaccine consisting of highly conserved outer membrane proteins (OMPs) may therefore hold the most promise for safely inducing protective immunity without the potential for inducing GBS. It is well recognized that C. jejuni strains are highly variable, and this will certainly impact on the development of a protein subunit vaccine. A protein appropriate for vaccine inclusion must be conserved in the largest possible proportion of C. jejuni strains, must be immunogenic, and must induce protective immunity against a large number of diverse C. jejuni strains. While analysis of the C. jejuni genome sequence is helpful as a starting point toward understanding its complement of OMPs, only direct identification of OMPs (by proteome analysis) will provide detailed information about the OMPs actually expressed by C. jejuni strains. Hypothesis: Certain C. jejuni outer membrane proteins (OMPs) will be conserved among all C. jejuni strains, will be immunogenic during human infection, and will generate a protective immune response. Specific Aim 1. We will identify the protein constituents of the outer membranes of several C. jejuni strains using proteomics and mass spectrometry. Specific Aim 2. We will determine whether OMPs that are conserved in our initial strains are also found in a large number of C. jejuni strains (fresh clinical isolates and an archival collection of strains from across the U.S.), and will evaluate the immune responses of infected humans to these OMPs. Specific Aim 3. We will determine whether immunization of mice with conserved, purified recombinant OMPs protects against subsequent experimental C. jejuni infection.