Structural implications of lipoarabinomannan glycans from global clinical isolates in diagnosis of Mycobacterium tuberculosis infection.
Structural implications of lipoarabinomannan glycans from global clinical isolates in diagnosis of Mycobacterium tuberculosis infection.
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DOI：
10.1016/j.jbc.2021.101265
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发表时间：
2021-11
期刊：
影响因子：
--
通讯作者：
Chatterjee D
中科院分区：
文献类型：
--
作者：
De P;Amin AG;Flores D;Simpson A;Dobos K;Chatterjee D
在结核分枝杆菌（Mtb）中，表面暴露的脂阿拉伯甘露聚糖（LAM）是免疫原性的关键决定因素，但其内在的异质性混淆了典型的结构-功能分析。最近，LAM作为活动性结核病（TB）感染的有效标志物获得了稳固的立足点，并在新的诊断工作中显示出巨大的潜力。然而，尚未做出努力来模拟或评估混合多克隆Mtb感染（用多种菌株感染）对除抗生素敏感性测试之外的TB诊断程序的影响。在这里，我们选择了三个TB临床分离株（HN 878，EAI和IO），并从这些菌株中纯化LAM，以提供一维和二维核磁共振（NMR）光谱，以及酶消化和位点特异性质谱（MS）的综合分析方法，以在多个水平上探测LAM的结构和行为。总体而言，我们发现所有LAM制剂中的聚糖相似，尽管存在细微变化。琥珀酸盐、乳酸盐、羟基丁酸盐、乙酸盐和Mtb LAM-甲硫基木糖（MTX）的标志物装饰了这些LAM物质的非还原性末端阿拉伯聚糖。新鉴定的乙酰氧基/羟基丁酸酯仅存在于来自EAI和IO Mtb菌株的LAM中。值得注意的是，详细的LC/MS-MS明确显示，LAM中的所有酰基修饰和乳基醚均位于2-连接的阿拉伯呋喃糖的3-OH位置，与末端β-阿拉伯呋喃糖相邻。最后，在LAM的连续酶促去糖基化后，连接有超过50% α-阿拉伯呋喃糖-（1→5）的残留聚糖不与单克隆抗体CS35结合。这些数据清楚地表明阿拉伯聚糖末端排列对LAM抗原性的重要性。
In Mycobacterium tuberculosis (Mtb), surface-exposed Lipoarabinomannan (LAM) is a key determinant of immunogenicity, yet its intrinsic heterogeneity confounds typical structure–function analysis. Recently, LAM gained a strong foothold as a validated marker for active tuberculosis (TB) infection and has shown great potential in new diagnostic efforts. However, no efforts have yet been made to model or evaluate the impact of mixed polyclonal Mtb infections (infection with multiple strains) on TB diagnostic procedures other than antibiotic susceptibility testing. Here, we selected three TB clinical isolates (HN878, EAI, and IO) and purified LAM from these strains to present an integrated analytical approach of one-dimensional and two-dimensional Nuclear Magnetic Resonance (NMR) spectroscopy, as well as enzymatic digestion and site-specific mass spectrometry (MS) to probe LAM structure and behavior at multiple levels. Overall, we found that the glycan was similar in all LAM preparations, albeit with subtle variations. Succinates, lactates, hydroxybutyrate, acetate, and the hallmark of Mtb LAM-methylthioxylose (MTX), adorned the nonreducing terminal arabinan of these LAM species. Newly identified acetoxy/hydroxybutyrate was present only in LAM from EAI and IO Mtb strains. Notably, detailed LC/MS-MS unambiguously showed that all acyl modifications and the lactyl ether in LAM are at the 3-OH position of the 2-linked arabinofuranose adjacent to the terminal β-arabinofuranose. Finally, after sequential enzymatic deglycosylation of LAM, the residual glycan that has ∼50% of α−arabinofuranose -(1→5) linked did not bind to monoclonal antibody CS35. These data clearly indicate the importance of the arabinan termini arrangements for the antigenicity of LAM.
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4.8
作者：
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通讯作者：
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DOI：
10.1097/qai.0000000000000672
发表时间：
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期刊：
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影响因子：
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