Detection of Mycobacterium tuberculosis peptides in the exosomes of patients with active and latent M. tuberculosis infection using MRM-MS.
Detection of Mycobacterium tuberculosis peptides in the exosomes of patients with active and latent M. tuberculosis infection using MRM-MS.
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DOI：
10.1371/journal.pone.0103811
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Dobos KM
中科院分区：
文献类型：
--
作者：
Kruh-Garcia NA;Wolfe LM;Chaisson LH;Worodria WO;Nahid P;Schorey JS;Davis JL;Dobos KM
鉴定活动性结核病易于测量、准确的诊断性生物标志物将对全球结核病控制工作产生重大影响。由于结核病发病机制涉及宿主和病原体的复杂性，确定一个足够敏感和特异性的单一生物标志物仍然是一个主要障碍。我们之前对结核病模型的研究表明，外泌体，如被感染巨噬细胞释放的外泌体，含有分枝杆菌产物，包括许多结核分枝杆菌蛋白。在本报告中，我们描述了利用多重反应监测质谱（MRM-MS）的靶向蛋白质组学分析的发展，以便我们能够跟踪以前通过western blot或霰弹枪质谱鉴定的蛋白质，并加强生物标志物的发现，包括检测人血清外泌体中的结核分枝杆菌蛋白。从培养确认的活动性结核病患者的血清样本中分离出外泌体，应用靶向MRM-MS检测76个肽，代表33种独特的结核分枝杆菌蛋白。我们的研究首次在人类血清外泌体中发现了细菌来源的活动性结核病生物标志物候选物。33种蛋白中有20种在结核病患者的外泌体中发现，包括来自8种蛋白（抗原85B、抗原85C、Apa、BfrB、GlcB、HspX、KatG和Mpt64）的多肽。有趣的是，所有这些蛋白质都是已知的分枝杆菌粘附素和/或有助于结核分枝杆菌细胞内存活的蛋白质。这些蛋白将作为靶分析物纳入未来的验证研究，因为它们可能作为持续活动性和潜伏性结核分枝杆菌感染的标记物。总之，这项工作是鉴定外泌体中封装的独特和特异性Mtb肽生物标志物的第一步，并揭示了结核病疾病状态谱中的复杂生物标志物模式。
The identification of easily measured, accurate diagnostic biomarkers for active tuberculosis (TB) will have a significant impact on global TB control efforts. Because of the host and pathogen complexities involved in TB pathogenesis, identifying a single biomarker that is adequately sensitive and specific continues to be a major hurdle. Our previous studies in models of TB demonstrated that exosomes, such as those released from infected macrophages, contain mycobacterial products, including many Mtb proteins. In this report, we describe the development of targeted proteomics assays employing multiplexed multiple reaction monitoring mass spectrometry (MRM-MS) in order to allow us to follow those proteins previously identified by western blot or shotgun mass spectrometry, and enhance biomarker discovery to include detection of Mtb proteins in human serum exosomes. Targeted MRM-MS assays were applied to exosomes isolated from human serum samples obtained from culture-confirmed active TB patients to detect 76 peptides representing 33 unique Mtb proteins. Our studies revealed the first identification of bacteria-derived biomarker candidates of active TB in exosomes from human serum. Twenty of the 33 proteins targeted for detection were found in the exosomes of TB patients, and included multiple peptides from 8 proteins (Antigen 85B, Antigen 85C, Apa, BfrB, GlcB, HspX, KatG, and Mpt64). Interestingly, all of these proteins are known mycobacterial adhesins and/or proteins that contribute to the intracellular survival of Mtb. These proteins will be included as target analytes in future validation studies as they may serve as markers for persistent active and latent Mtb infection. In summary, this work is the first step in identifying a unique and specific panel of Mtb peptide biomarkers encapsulated in exosomes and reveals complex biomarker patterns across a spectrum of TB disease states.
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DOI：
10.1097/qai.0b013e3181eb611a
发表时间：
2010-12
期刊：
Journal of acquired immune deficiency syndromes (1999)
影响因子：
--
作者：
Kyeyune R;den Boon S;Cattamanchi A;Davis JL;Worodria W;Yoo SD;Huang L
通讯作者：
Huang L
影响因子：
5.2
作者：
Forrellad MA;Klepp LI;Gioffré A;Sabio y García J;Morbidoni HR;de la Paz Santangelo M;Cataldi AA;Bigi F
通讯作者：
Bigi F
影响因子：
3.8
作者：
King HW;Michael MZ;Gleadle JM
通讯作者：
Gleadle JM
影响因子：
3.2
作者：
Izzo, A;Brandt, L;Orme, I
通讯作者：
Orme, I
影响因子：
46.9
作者：
Addona, Terri A.;Shi, Xu;Keshishian, Hasmik;Mani, D. R.;Burgess, Michael;Gillette, Michael A.;Clauser, Karl R.;Shen, Dongxiao;Lewis, Gregory D.;Farrell, Laurie A.;Fifer, Michael A.;Sabatine, Marc S.;Gerszten, Robert E.;Carr, Steven A.
通讯作者：
Carr, Steven A.