Ultra-dense peptide array analysis of naturally acquired and vaccine-induced P. falciparum immunity

项目摘要 一种有效的血液阶段恶性疟原虫疟疾疫苗将对全球健康产生巨大的益处， 特别是对于遭受疟疾发病率和死亡率最大负担的婴儿和幼儿。 我们对疫苗诱导的和自然获得的免疫保护相关性的知识有限 这是开发这种疫苗的主要障碍。阐明B细胞表位可以 提供疫苗研制所需的信息。高密度肽微阵列将提供 这意味着检查抗原多样性的广度和深度，以及这种多样性在免疫系统中的作用。 保护性免疫力的发展，我们希望用疫苗来模仿。此外，更高的精度 比目前可用的，以及相应的详细的表位作图，将是可能的， 创建了一个密集重叠的肽阵列，完全覆盖了恶性疟原虫蛋白质组， 肽组。我们建议研究自然获得的和疫苗诱导的疟疾免疫， 16-mer肽序列的肽微阵列，其总体目标是鉴定与这些肽相关的肽， 提供保护并为疟疾疫苗的开发提供信息。我们将使用一种新技术， 成功用于人肽组，以研究人对寄生虫衍生肽的血清反应性， 首次我们将创建两个阵列：1）多样性阵列将包含代表以下的短柔性肽： 基于疟原虫测序的疟疾疫苗候选抗原及其不同变体 2）肽组阵列将涵盖对应于孔的所有恶性疟原虫肽。 特征参考菌株。这两个阵列将提供用于精细表位作图和 鉴定自然获得的和疫苗诱导的保护相关物。相应抗原 作为潜在的疫苗值得进一步研究。从该项目中获得的知识将使 超密集肽阵列可用于研究对其他感染性生物体的体液免疫应答。

PROJECT SUMMARY An effective blood stage Plasmodium falciparum malaria vaccine would be of immense global health benefit, particularly for infants and young children who suffer the greatest burden of malaria morbidity and mortality. Our limited knowledge of vaccine-induced and naturally acquired correlates of immune protection represents a major impediment to the development of such a vaccine. Elucidating B cell epitopes may provide information needed for vaccine development. A high-density peptide microarray would provide the means to examine both the breadth and depth of antigenic diversity and the role of this diversity in the development of protective immunity that we wish to emulate with a vaccine. Furthermore, finer precision than what is currently available, and correspondingly detailed epitope mapping, will be possible through the creation a densely overlapping peptide array with full coverage of the falciparum proteome, a so-called peptidome. We propose to study naturally acquired and vaccine-induced immunity to malaria with novel peptide microarrays of 16-mer peptide sequences with the overall goals of identifying peptides that correlate with protection and informing malaria vaccine development. We will use a new technology that has been used successfully for a human peptidome to study human seroreactivity to parasite-derived peptides for the first time. We will create two arrays: 1) A diversity array will contain short flexible peptides that represent malaria vaccine candidate antigens and their diverse variants based on sequencing of malaria parasites from the field; and 2) A peptidome array will encompass all P. falciparum peptides corresponding to a well- characterized reference strain. These two arrays will provide the means for both fine epitope mapping and identification of naturally acquired and vaccine-induced correlates of protection. The corresponding antigens will be worthy of further study as potential vaccines. Knowledge gained from this project will enable the ultra-dense peptide array to be used to study the humoral immune response to other infectious organisms.