Fasciola hepatica Extracellular Vesicles isolated from excretory-secretory products using a gravity flow method modulate dendritic cell phenotype and activity.
Fasciola hepatica Extracellular Vesicles isolated from excretory-secretory products using a gravity flow method modulate dendritic cell phenotype and activity.
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DOI：
10.1371/journal.pntd.0008626
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发表时间：
2020-09
影响因子：
3.8
通讯作者：
O'Neill SM
中科院分区：
文献类型：
--
作者：
Murphy A;Cwiklinski K;Lalor R;O'Connell B;Robinson MW;Gerlach J;Joshi L;Kilcoyne M;Dalton JP;O'Neill SM
寄生虫释放的细胞外囊泡（EV）向宿主免疫系统传递信号，这对维持寄生虫和宿主之间的长期关系至关重要。在本研究中，总EV（FhEV）释放的蠕虫寄生虫肝片吸虫成虫体外分离使用最近描述的重力流方法，保护其结构的完整性。使用蛋白质组学分析和通过聚糖微阵列表征的其表面拓扑结构来定义FhEV分子货物。蛋白质组学分析确定了618个蛋白质，其中121个含有推定的N-连接的糖基化位点，而132个蛋白质含有推定的O-连接的糖基化位点。聚糖阵列显示表面暴露的聚糖与甘露糖结合凝集素的高亲和力，表明寡甘露糖丰富的糖蛋白的优势，以及其他聚糖与复合型N-聚糖的高亲和力。当添加到骨髓来源的树突状细胞分离FhEV诱导一种新的表型，其分类为分泌低水平的TNF，细胞表面标志物（CD 80，CD 86，CD 40，OX 40 L和SIGNR 1）的表达增强和细胞内标志物（SOCS 1和SOCS 3）的升高。当通过过继转移将FhEV刺激的BMDC引入OT-II小鼠时，特异性和非特异性抗原刺激均抑制皮肤引流淋巴结和脾细胞的IL-2分泌。用FhEV悬浮液免疫小鼠未引起显著的免疫应答;然而，在明矾存在下，FhEV诱导具有高抗原特异性抗体滴度的混合Th 1/Th 2免疫应答。因此，我们已经证明FhEV在DC中诱导能够抑制IL-2从T细胞分泌的独特表型。我们的研究增加了不断增长的免疫蛋白质组学数据库，这将是发现未来寄生虫疫苗和免疫生物制剂的重要来源。寄生虫释放的细胞外囊泡（EV）向宿主免疫系统传递信号，这对维持寄生虫和宿主之间的长期关系至关重要。本研究使用保护囊泡结构完整性的重力流方法分离由寄生虫肝片吸虫成虫阶段在体外释放的总EV（FhEV）。蛋白质组学分析鉴定了618种蛋白质，其中121种含有推定的N-连接糖基化位点，而132种蛋白质含有推定的O-连接糖基化位点，而聚糖阵列显示表面暴露的聚糖主要是富含寡聚甘露糖的糖蛋白，并且聚糖对复合型N-聚糖具有高亲和力。由于EV分子货物可以影响宿主免疫细胞，因此将FhEV添加到骨髓来源的树突状细胞中，诱导一种新的细胞表型，当过继转移到OT-II小鼠中时，该表型会抑制皮肤引流淋巴结和脾细胞分泌IL-2。用FhEV免疫小鼠没有引起显著的免疫应答;然而，在明矾的存在下，FhEV诱导具有高抗原特异性抗体滴度的混合Th 1/Th 2免疫应答。这项研究揭示了FhEV的生物活性，并添加到不断增长的免疫蛋白质组学数据库中，这将成为发现未来治疗方法的重要来源。
Parasite-released extracellular vesicles (EVs) deliver signals to the host immune system that are critical to maintaining the long-term relationship between parasite and host. In the present study, total EVs (FhEVs) released in vitro by adults of the helminth parasite Fasciola hepatica were isolated using a recently described gravity flow method that protects their structural integrity. The FhEVs molecular cargo was defined using proteomic analysis and their surface topology characterised by glycan microarrays. The proteomic analysis identified 618 proteins, 121 of which contained putative N-linked glycosylation sites while 132 proteins contained putative O-linked glycosylation sites. Glycan arrays revealed surface-exposed glycans with a high affinity for mannose-binding lectins indicating the predominance of oligo mannose-rich glycoproteins, as well as other glycans with a high affinity for complex-type N-glycans. When added to bone-marrow derived dendritic cells isolated FhEV induced a novel phenotype that was categorised by the secretion of low levels of TNF, enhanced expression of cell surface markers (CD80, CD86, CD40, OX40L, and SIGNR1) and elevation of intracellular markers (SOCS1 and SOCS3). When FhEV-stimulated BMDCs were introduced into OT-II mice by adoptive transfer, IL-2 secretion from skin draining lymph nodes and spleen cells was inhibited in response to both specific and non-specific antigen stimulation. Immunisation of mice with a suspension of FhEV did not elicit significant immune responses; however, in the presence of alum, FhEVs induced a mixed Th1/Th2 immune response with high antigen specific antibody titres. Thus, we have demonstrated that FhEVs induce a unique phentotype in DC capable of suppressing IL-2 secretion from T-cells. Our studies add to the growing immuno-proteomic database that will be an important source for the discovery of future parasite vaccines and immunotherapeutic biologicals. Parasite-released extracellular vesicles (EVs) deliver signals to the host immune system that are critical to maintaining the long-term relationship between parasite and host. This study isolated total EVs (FhEVs) released in vitro by the adult stages of the parasitic worm Fasciola hepatica using a gravity flow method that protects the structural integrity of the vesicles. Proteomic analysis identified 618 proteins, 121 of which contained putative N-linked glycosylation sites while 132 proteins contained putative O-linked glycosylation sites while glycan arrays revealed surface-exposed glycans were predominantly oligo mannose-rich glycoproteins, and glycans with a high affinity for complex-type N-glycans. Since the EV molecular cargo can influence host immune cells, FhEVs were added to bone-marrow derived dendritic cells, inducing a novel cell phenotype that when adoptive transferred into OT-II mice inhibited IL-2 secretion from skin draining lymph nodes and spleen cells. Immunisation of mice with FhEV did not elicit significant immune responses; however, in the presence of alum, FhEVs induced a mixed Th1/Th2 immune response with high antigen specific antibody titres. This studied sheds like on the biological activity of FhEVs and added to the growing immuno-proteomic database that will be an important source for the discovery of future therapeutics.
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DOI：
10.1093/infdis/jiv291
发表时间：
2015-11-15
期刊：
The Journal of infectious diseases
影响因子：
--
作者：
Chaiyadet S;Sotillo J;Smout M;Cantacessi C;Jones MK;Johnson MS;Turnbull L;Whitchurch CB;Potriquet J;Laohaviroj M;Mulvenna J;Brindley PJ;Bethony JM;Laha T;Sripa B;Loukas A
通讯作者：
Loukas A
影响因子：
16
作者：
Chivet M;Javalet C;Laulagnier K;Blot B;Hemming FJ;Sadoul R
通讯作者：
Sadoul R
影响因子：
6.1
作者：
Gatti, Stefano;Bruno, Stefania;Camussi, Giovanni
通讯作者：
Camussi, Giovanni
影响因子：
2.1
作者：
Azizi, Hakim;Mirzaeei, Hadi;Yaghoobi, Hajar
通讯作者：
Yaghoobi, Hajar
影响因子：
1.8
作者：
Cervi, L;Borgonovo, J;Masih, D
通讯作者：
Masih, D