Felgner Philip L, Kayala Matthew A, Vigil Adam, Burk Chad, Nakajima-Sasaki Rie, Pablo Jozelyn, Molina Douglas M, Hirst Siddiqua, Chew Janet S W, Wang Dongling, Tan Gladys, Duffield Melanie, Yang Ron, Neel Julien, Chantratita Narisara, Bancroft Greg, Lertmemongkolchai Ganjana, Davies D Huw, Baldi Pierre, Peacock Sharon, Titball Richard W
Department of Medicine, Division of Infectious Diseases, University of California, Irvine, CA 92697, USA.
Proc Natl Acad Sci U S A. 2009 Aug 11;106(32):13499-504. doi: 10.1073/pnas.0812080106. Epub 2009 Jul 28.
Understanding the way in which the immune system responds to infection is central to the development of vaccines and many diagnostics. To provide insight into this area, we fabricated a protein microarray containing 1,205 Burkholderia pseudomallei proteins, probed it with 88 melioidosis patient sera, and identified 170 reactive antigens. This subset of antigens was printed on a smaller array and probed with a collection of 747 individual sera derived from 10 patient groups including melioidosis patients from Northeast Thailand and Singapore, patients with different infections, healthy individuals from the USA, and from endemic and nonendemic regions of Thailand. We identified 49 antigens that are significantly more reactive in melioidosis patients than healthy people and patients with other types of bacterial infections. We also identified 59 cross-reactive antigens that are equally reactive among all groups, including healthy controls from the USA. Using these results we were able to devise a test that can classify melioidosis positive and negative individuals with sensitivity and specificity of 95% and 83%, respectively, a significant improvement over currently available diagnostic assays. Half of the reactive antigens contained a predicted signal peptide sequence and were classified as outer membrane, surface structures or secreted molecules, and an additional 20% were associated with pathogenicity, adaptation or chaperones. These results show that microarrays allow a more comprehensive analysis of the immune response on an antigen-specific, patient-specific, and population-specific basis, can identify serodiagnostic antigens, and contribute to a more detailed understanding of immunogenicity to this pathogen.
了解免疫系统对感染的反应方式是疫苗和许多诊断方法研发的核心。为了深入了解这一领域，我们制作了一个包含1205种伯克霍尔德菌蛋白的蛋白质微阵列，用88份类鼻疽病患者血清对其进行检测，并鉴定出170种反应性抗原。将这一抗原子集印在一个较小的阵列上，并用来自10个患者群体的747份个体血清进行检测，这些群体包括来自泰国东北部和新加坡的类鼻疽病患者、患有不同感染的患者、来自美国的健康个体以及来自泰国地方病和非地方病地区的个体。我们鉴定出49种在类鼻疽病患者中比健康人和其他类型细菌感染患者反应性显著更高的抗原。我们还鉴定出59种在所有群体（包括来自美国的健康对照）中反应性相同的交叉反应性抗原。利用这些结果，我们设计出一种检测方法，该方法能够分别以95%的灵敏度和83%的特异性对类鼻疽病阳性和阴性个体进行分类，这比目前可用的诊断检测方法有显著改进。一半的反应性抗原含有预测的信号肽序列，被归类为外膜、表面结构或分泌分子，另外20%与致病性、适应性或伴侣蛋白相关。这些结果表明，微阵列能够在抗原特异性、患者特异性和群体特异性基础上对免疫反应进行更全面的分析，能够鉴定血清诊断抗原，并有助于更详细地了解对该病原体的免疫原性。
