Identification of Biomarkers for Systemic Distribution of Nanovesicles From Lactobacillus johnsonii N6.2.
Identification of Biomarkers for Systemic Distribution of Nanovesicles From Lactobacillus johnsonii N6.2.
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DOI：
10.3389/fimmu.2021.723433
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发表时间：
2021
影响因子：
7.3
通讯作者：
Lorca GL
中科院分区：
文献类型：
--
作者：
Harrison NA;Gardner CL;da Silva DR;Gonzalez CF;Lorca GL
细菌细胞外囊泡（EV）转运生物分子的能力增加了确定其作为治疗剂的潜力的研究。在这项研究中，对约氏乳杆菌N6.2衍生的纳米囊泡（NV）进行了表征，以鉴定可作为宿主-微生物相互作用中生物标志物的组分。比较蛋白质组学和脂质组学分析。约氏N6.2 NV和细胞膜（CM）。脂质组学图谱表明，这两种组分含有相似的脂质，然而，在几个类别中观察到显着差异。LC-MS/MS蛋白质组学分析表明，NV含有几个独特的和差异表达的蛋白质相比，CM。基于细胞组分的基因本体（GO）术语的分析显示NV级分的蛋白质在细胞质/细胞内空间类别中显著富集。基于这些结果，选择蛋白T285_RS00825（命名为Sdp）、Eno 3和莱克萨用于定位研究并作为宿主-微生物相互作用的潜在生物标志物。免疫金染色，然后通过扫描和透射电子显微镜（SEM和TEM，分别），显示SDP优先定位沿着细胞壁/膜，并在NV样结构周围的细菌。这些结果在用NV孵育的Caco-2细胞中使用免疫荧光染色证实。因此，我们评估了NV表面暴露蛋白在宿主中产生免疫应答的潜力。来自施用L. johnsonii N6.2的结果表明，在体内产生了针对NV和Sdp结构域的伊加和IgG抗体。这些结果表明L. johnsonii N6.2 NV具有通过免疫调节介导宿主相互作用的潜力。
The ability of bacterial extracellular vesicles (EV) to transport biological molecules has increased the research to determine their potential as therapeutic agents. In this study, Lactobacillus johnsonii N6.2-derived nanovesicles (NV) were characterized to identify components that may serve as biomarkers in host-microbe interactions. Comparative proteomic and lipidomic analyses of L. johnsonii N6.2 NV and cell membrane (CM) were performed. The lipidomic profiles indicated that both fractions contained similar lipids, however, significant differences were observed in several classes. LC-MS/MS proteomic analysis indicated that NV contained several unique and differentially expressed proteins when compared to the CM. Analysis of Gene Ontology (GO) terms, based on cellular component, showed significant enrichment of proteins in the cytoplasm/intracellular space category for the NV fraction. Based on these results, the proteins T285_RS00825 (named Sdp), Eno3 and LexA were selected for studies of localization and as potential biomarkers for host-microbe interactions. Immunogold staining, followed by scanning and transmission electron microscopy (SEM and TEM, respectively), revealed that Sdp was preferentially localized along the cell wall/membrane, and on NV-like structures surrounding the bacteria. These results were confirmed using immunofluorescence staining in Caco-2 cells incubated with NV. Consequently, we evaluated the potential for NV surface-exposed proteins to generate an immune response in the host. Plasma from individuals administered L. johnsonii N6.2 showed that IgA and IgG antibodies were generated against NV and Sdp domains in vivo. Altogether, these results show that L. johnsonii N6.2 NV have the potential to mediate host interactions through immune modulation.