Immunoproteomic Analysis of Antibody in Lymphocyte Supernatant in Patients with Typhoid Fever in Bangladesh
Immunoproteomic Analysis of Antibody in Lymphocyte Supernatant in Patients with Typhoid Fever in Bangladesh
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DOI：
10.1128/cvi.00661-13
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发表时间：
2014-03-01
影响因子：
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通讯作者：
Ryan, Edward T.
中科院分区：
文献类型：
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作者：
Charles, Richelle C.;Liang, Li;Ryan, Edward T.
我们之前已经证明，基于检测伤寒患者淋巴细胞上清液中的抗肠道沙门氏菌血清型伤寒抗体的检测方法（淋巴细胞上清液中的抗体[ALS]检测）可以识别孟加拉国100%的血培养确诊伤寒患者。为了确定S.伤寒膜制备物在这些先前的研究中用作抗原，并鉴定了伤寒沙门氏菌特有的潜在生物标志物。伤寒菌血症患者，我们探测含有2,724个S的微阵列。伤寒蛋白与ALS收集在临床表现时，从10名孟加拉国急性伤寒。我们确定了62个免疫反应性抗原时，评价IgG和伊加的反应。对这些抗原中的10种的免疫反应区分了急性伤寒感染的个体和来自伤寒感染流行地区的健康对照个体，以及随后被证实患有非伤寒病的发热孟加拉国患者。使用ALS酶联免疫吸附试验（ELISA）的形式和纯化的抗原，然后，我们证实，对抗原的免疫反应性最高（溶血素E [HlyE]）正确识别个体急性伤寒或副伤寒在达卡，孟加拉国。这些观察结果表明，纯化的抗原可以与ALS和相应的急性期活化的B淋巴细胞一起用于诊断平台，以鉴定急性感染的患者，甚至在肠热病流行的地区。
We have previously shown that an assay based on detection of anti-Salmonella enterica serotype Typhi antibodies in supernatant of lymphocytes harvested from patients presenting with typhoid fever (antibody in lymphocyte supernatant [ALS] assay) can identify 100% of patients with blood culture-confirmed typhoid fever in Bangladesh. In order to define immunodominant proteins within the S. Typhi membrane preparation used as antigen in these prior studies and to identify potential biomarkers unique to S. Typhi bacteremic patients, we probed microarrays containing 2,724 S. Typhi proteins with ALS collected at the time of clinical presentation from 10 Bangladeshis with acute typhoid fever. We identified 62 immunoreactive antigens when evaluating both the IgG and IgA responses. Immune responses to 10 of these antigens discriminated between individuals with acute typhoid infection and healthy control individuals from areas where typhoid infection is endemic, as well as Bangladeshi patients presenting with fever who were subsequently confirmed to have a nontyphoid illness. Using an ALS enzyme-linked immunosorbent assay (ELISA) format and purified antigen, we then confirmed that immune responses against the antigen with the highest immunoreactivity (hemolysin E [HlyE]) correctly identified individuals with acute typhoid or paratyphoid fever in Dhaka, Bangladesh. These observations suggest that purified antigens could be used with ALS and corresponding acute-phase activated B lymphocytes in diagnostic platforms to identify acutely infected patients, even in areas where enteric fever is endemic.