A systemic approach to identify non-abundant immunogenic proteins in Lyme disease pathogens.
A systemic approach to identify non-abundant immunogenic proteins in Lyme disease pathogens.
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DOI：
10.1128/msystems.01087-23
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发表时间：
2024-01-23
期刊：
影响因子：
6.4
通讯作者：
Pal, Utpal
中科院分区：
文献类型：
--
作者：
Yas, Ozlem Buyuktanir;Coleman, Adam S.;Lipman, Rachel M.;Sharma, Kavita;Raghunandanan, Sajith;Alanazi, Fuad;Rana, Vipin S.;Kitsou, Chrysoula;Yang, Xiuli;Pal, Utpal
莱姆病病原体伯氏疏螺旋体（Borrelia burgdorferi）差异性地产生许多外表面蛋白（Osp），其中一些代表最丰富的膜蛋白，如OspA、OspB和OspC。在培养的细菌中，这些蛋白质可以占总细胞或膜蛋白的相当大的部分，这对非丰富蛋白质的鉴定和分析构成了挑战，这些蛋白质可以作为新的病原体检测标记物或作为疫苗候选物。在此，我们引入连续突变以去除这些丰富的Osps并产生B。在感染性297分离株背景中，存在OspA、OspB和OspC缺陷的伯氏螺旋体突变体，命名为OspABC−突变体。与亲本分离株相比，突变体没有反映在培养基中的生长缺陷，但表现出差异的mRNA表达的代表性测试基因，除了细胞和膜蛋白谱的总变化。通过二维凝胶电泳和液相色谱-质谱法对OspABC−突变体与野生型相比差异可检测的蛋白质含量进行分析，确定了几种螺旋体蛋白质，这些蛋白质主要由未知功能的蛋白质以及膜转运蛋白、伴侣蛋白和代谢酶组成。我们生产了这些代表性蛋白质中的两种的重组形式，BBA 34和BB 0238，并且表明这些蛋白质在蜱传的莱姆疏螺旋体病小鼠模型中的螺旋体感染期间是可检测的，并且因此作为感染的潜在抗原标志物。本手稿采用系统性方法来鉴定培养的伯氏疏螺旋体中的非丰富蛋白质，这些蛋白质由于大量存在OspA、OspB和OspC等Osps而被掩盖或隐藏。由于这些Osps在哺乳动物中不存在或瞬时表达，我们进行了概念验证研究，其中它们的去除允许分析OspABC缺陷型突变体中原本丰度较低的抗原，并鉴定了几种免疫原性蛋白，包括BBA 34和BB 0238。这些抗原可作为莱姆病的新候选疫苗和/或遗传标记，促进莱姆病临床诊断和预防的新研究。
Borrelia burgdorferi, the pathogen of Lyme disease, differentially produces many outer surface proteins (Osp), some of which represent the most abundant membrane proteins, such as OspA, OspB, and OspC. In cultured bacteria, these proteins can account for a substantial fraction of the total cellular or membrane proteins, posing challenges to the identification and analysis of non-abundant proteins, which could serve as novel pathogen detection markers or as vaccine candidates. Herein, we introduced serial mutations to remove these abundant Osps and generated a B. burgdorferi mutant deficient in OspA, OspB, and OspC in an infectious 297-isolate background, designated as OspABC− mutant. Compared to parental isolate, the mutant did not reflect growth defects in the cultured medium but showed differential mRNA expression of representative tested genes, in addition to gross changes in cellular and membrane protein profiles. The analysis of differentially detectable protein contents of the OspABC− mutant, as compared to the wild type, by two-dimensional gel electrophoresis followed by liquid chromatography-mass spectrometry, identified several spirochete proteins that are dominated by proteins of unknown functions, as well as membrane transporters, chaperons, and metabolic enzymes. We produced recombinant forms of two of these represented proteins, BBA34 and BB0238, and showed that these proteins are detectable during spirochete infection in the tick-borne murine model of Lyme borreliosis and thus serve as potential antigenic markers of the infection. The present manuscript employed a systemic approach to identify non-abundant proteins in cultured Borrelia burgdorferi that are otherwise masked or hidden due to the overwhelming presence of abundant Osps like OspA, OspB, and OspC. As these Osps are either absent or transiently expressed in mammals, we performed a proof-of-concept study in which their removal allowed the analysis of otherwise less abundant antigens in OspABC-deficient mutants and identified several immunogenic proteins, including BBA34 and BB0238. These antigens could serve as novel vaccine candidates and/or genetic markers of Lyme borreliosis, promoting new research in the clinical diagnosis and prevention of Lyme disease.
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影响因子：
3.6
作者：
He M;Oman T;Xu H;Blevins J;Norgard MV;Yang XF
通讯作者：
Yang XF
DOI：
10.1084/jem.20070962
发表时间：
2008-01-21
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Pal U;Wang P;Bao F;Yang X;Samanta S;Schoen R;Wormser GP;Schwartz I;Fikrig E
通讯作者：
Fikrig E
影响因子：
3.7
作者：
Adrion ER;Aucott J;Lemke KW;Weiner JP
通讯作者：
Weiner JP
影响因子：
11.8
作者：
Gomes-Solecki, Maria;Arnaboldi, Paul M.;Schutzer, Steven E.
通讯作者：
Schutzer, Steven E.
影响因子：
4.4
作者：
Mead, Paul S.
通讯作者：
Mead, Paul S.