Protrusion-guided extracellular vesicles mediate CD30 trans-signalling in the microenvironment of Hodgkin's lymphoma
Protrusion-guided extracellular vesicles mediate CD30 trans-signalling in the microenvironment of Hodgkin's lymphoma
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DOI：
10.1002/path.4306
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发表时间：
2014-03-01
影响因子：
7.3
通讯作者：
von Strandmann, Elke Pogge
中科院分区：
文献类型：
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作者：
Hansen, Hinrich P.;Engels, Hanna-Mareike;von Strandmann, Elke Pogge
经典霍奇金淋巴瘤（cHL）受累淋巴组织仅含有少量恶性霍奇金和里德-斯滕伯格（HRS）细胞，这些细胞在大量反应性细胞浸润中播散。特别地，先天免疫浸润被认为通过直接的细胞-细胞相互作用支持肿瘤生长。由于它们很少在原位与恶性细胞紧密接触，我们研究了cHL衍生的细胞外囊泡是否可以替代直接的细胞-细胞接触。我们研究了跨膜蛋白CD 30和CD 30配体（CD 30 L）的串扰，因为它们分别在HRS和先天免疫细胞上选择性表达。在这里，我们发现HRS细胞释放胞外域作为可溶性分子（sCD 30）和细胞外囊泡表面上的整个受体。通过冷冻和免疫电子显微镜测定，囊泡直径为40-800 nm。除CD 30外，通过质谱和流式细胞术检测典型的细胞外囊泡标志物，包括四跨膜蛋白、漂浮蛋白、热休克蛋白和粘附分子。与sCD 30相反，囊泡在来自健康供体的CD 30 L+嗜酸性粒细胞样EoL-1细胞和原代粒细胞中引起CD 30依赖性白细胞介素-8释放，强调了囊泡上CD 30的功能。在HRS细胞的细胞外基质（ECM）包埋培养物中，基于肌动蛋白和微管蛋白的突起网络引导CD 30+囊泡进入微环境。该网络将CD 30+囊泡靶向远端免疫细胞，并引起CD 30 L的强烈极化。30 µm切片的共聚焦激光扫描显微镜显示，在混合细胞和结节硬化亚型的cHL受累淋巴组织中也存在含CD 30囊泡的网络。该网络可能促进cHL组织中远端细胞类型之间的通信，并允许功能性CD 30-CD 30 L相互作用。该网络的微管蛋白骨架可能为基于抗微管蛋白的CD 30抗体构建体治疗cHL提供靶点。版权所有© 2013大不列颠及爱尔兰病理学会。出版社：John Wiley & Sons，Ltd
Classical Hodgkin's lymphoma (cHL)‐affected lymphoid tissue contains only a few malignant Hodgkin and Reed–Sternberg (HRS) cells, which are disseminated within a massive infiltrate of reactive cells. In particular, the innate immune infiltrate is deemed to support tumour growth by direct cell–cell interaction. Since they are rarely found in close proximity to the malignant cellsin situ, we investigated whether cHL‐derived extracellular vesicles might substitute for a direct cell–cell contact. We studied the crosstalk of the transmembrane proteins CD30 and CD30 ligand (CD30L) because they are selectively expressed on HRS and innate immune cells, respectively. Here, we showed that HRS cells released both the ectodomain as a soluble molecule (sCD30) and the entire receptor on the surface of extracellular vesicles. The vesicle diameter was 40–800 nm, as determined by cryo‐ and immune electron microscopy. In addition to CD30, typical extracellular vesicle markers were detected by mass spectrometry and flow cytometry, including tetraspanins, flotillins, heat shock proteins and adhesion molecules. In contrast to sCD30, vesicles caused a CD30‐dependent release of interleukin‐8 in CD30L+eosinophil‐like EoL‐1 cells and primary granulocytes from healthy donors, underscoring the functionality of CD30 on vesicles. In extracellular matrix (ECM)‐embedded culture of HRS cells, a network of actin and tubulin‐based protrusions guided CD30+vesicles into the micro‐environment. This network targeted CD30+vesicles towards distant immune cells and caused a robust polarization of CD30L. Confocal laser scanning microscopy of 30 µm sections showed a CD30 vesicle‐containing network also in cHL‐affected lymphoid tissue of both mixed‐cellularity and nodular sclerosing subtypes. This network might facilitate the communication between distant cell types in cHL tissue and allow a functional CD30–CD30L interactionin trans. The tubulin backbone of the network may provide a target for the therapy of cHL with antitubulin‐based CD30 antibody constructs. Copyright © 2013 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.