Profiling humoral immune responses to P. falciparum infection with protein microarrays.
Profiling humoral immune responses to P. falciparum infection with protein microarrays.
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DOI：
10.1002/pmic.200800194
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发表时间：
2008-11
期刊：
影响因子：
3.4
通讯作者：
Felgner, Philip L.
中科院分区：
文献类型：
--
作者：
Doolan, Denise L.;Mu, Yunxiang;Unal, Berkay;Sundaresh, Suman;Hirst, Siddiqua;Valdez, Conrad;Randall, Arlo;Molina, Douglas;Liang, Xiaowu;Freilich, Daniel A.;Oloo, J. Aggrey;Blair, Peter L.;Aguiar, Joao C.;Baldi, Pierre;Davies, D. Huw;Felgner, Philip L.
缺乏对人类暴露于复杂病原体后的血清学反应的完整描述，适合实现这一点的方法也有限。在这里，我们报告了一种以疟疾为模型的方法，该方法阐明了在自然或实验感染后或在用减毒生物接种后产生的抗体的特征，并识别了疫苗开发或其他应用中感兴趣的免疫活性抗原。通过聚合酶链式反应/重组克隆构建了250个恶性疟原虫(Pf)蛋白的表达载体，在体外无细胞转录和翻译反应中以90%的效率分别表达恶性疟原虫(Pf)蛋白，不经纯化直接打印到芯片玻片上。蛋白质芯片用免疫状态不同的四组人血清之一进行探测：无菌免疫或没有对实验攻击的免疫力，通过辐射减毒的PF子孢子接种，通过自然暴露获得部分免疫，以及以前没有接触过PF。总体而言，共鉴定出72种高反应性的PF抗原。鉴定了与免疫反应性相关的蛋白质组学特征。重要的是，每个捐赠者的抗体谱都是不同的。从这种分析中获得的信息将有助于确定疫苗开发的抗原，剖析免疫的分子基础，监测全生物疫苗试验的结果，并确定与保护有关的免疫因素。
A complete description of the serological response following exposure of humans to complex pathogens is lacking and approaches suitable for accomplishing this are limited. Here we report, using malaria as a model, a method which elucidates the profile of antibodies that develop after natural or experimental infection or after vaccination with attenuated organisms, and which identifies immunoreactive antigens of interest for vaccine development or other applications. Expression vectors encoding 250 Plasmodium falciparum (Pf) proteins were generated by PCR/recombination cloning; the proteins were individually expressed with >90% efficiency in E. coli cell-free in vitro transcription and translation reactions, and printed directly without purification onto microarray slides. The protein microarrays were probed with human sera from one of four groups which differed in immune status: sterile immunity or no immunity against experimental challenge following vaccination with radiation-attenuated Pf sporozoites, partial immunity acquired by natural exposure, and no previous exposure to Pf. Overall, 72 highly reactive Pf antigens were identified. Proteomic features associated with immunoreactivity were identified. Importantly, antibody profiles were distinct for each donor group. Information obtained from such analyses will facilitate identifying antigens for vaccine development, dissecting the molecular basis of immunity, monitoring the outcome of whole-organism vaccine trials, and identifying immune correlates of protection.
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影响因子：
6.4
作者：
BEADLE, C;MCELROY, PD;HOFFMAN, SL
通讯作者：
HOFFMAN, SL
影响因子：
9.8
作者：
Bozdech, Zbynek;Llinas, Manuel;Pulliam, Brian Lee;Wong, Edith D;Zhu, Jingchun;DeRisi, Joseph L
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DeRisi, Joseph L
影响因子：
5.4
作者：
Benhnia, Mohammed Rafii-El-Idrissi;McCausland, Megan M.;Crotty, Shane
通讯作者：
Crotty, Shane
影响因子：
5.4
作者：
Davies, D. Huw;Wyatt, Linda S.;Felgner, Philip L.
通讯作者：
Felgner, Philip L.
影响因子：
64.8
作者：
HILL, AVS;ELVIN, J;WHITTLE, HC
通讯作者：
WHITTLE, HC