title: Protrusion-guided extracellular vesicles mediate CD30 trans-signalling in the microenvironment of Hodgkin's lymphoma.
abstract: 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 cells in 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 interaction in trans. The tubulin backbone of the network may provide a target for the therapy of cHL with antitubulin-based CD30 antibody constructs.
authors.affiliation: Department of Internal Medicine I, University of Cologne, Germany.
authors.forename: Hinrich P
authors.initials: HP
authors.lastname: Hansen
authors.forename: Hanna-Mareike
authors.initials: HM
authors.lastname: Engels
authors.forename: Maria
authors.initials: M
authors.lastname: Dams
authors.forename: Adriana F
authors.initials: AF
authors.lastname: Paes Leme
authors.forename: Bianca Alves
authors.initials: BA
authors.lastname: Pauletti
authors.forename: Vijaya Lakshmi
authors.initials: VL
authors.lastname: Simhadri
authors.forename: Horst
authors.initials: H
authors.lastname: Dürkop
authors.forename: Katrin S
authors.initials: KS
authors.lastname: Reiners
authors.forename: Sabine
authors.initials: S
authors.lastname: Barnert
authors.forename: Andreas
authors.initials: A
authors.lastname: Engert
authors.forename: Rolf
authors.initials: R
authors.lastname: Schubert
authors.forename: Fabio
authors.initials: F
authors.lastname: Quondamatteo
authors.forename: Michael
authors.initials: M
authors.lastname: Hallek
authors.forename: Elke
authors.initials: E
authors.lastname: Pogge von Strandmann
mesh_terms: D014408:Biomarkers, Tumor / Q000378:metabolism; D053285:CD30 Ligand / Q000378:metabolism; D002450:Cell Communication; D045744:Cell Line, Tumor; D022081:Cell Surface Extensions / Q000276:immunology / Q000378:metabolism* / Q000648:ultrastructure; D020285:Cryoelectron Microscopy; D004804:Eosinophils / Q000276:immunology / Q000378:metabolism; D005434:Flow Cytometry; D006098:Granulocytes / Q000276:immunology / Q000378:metabolism; D006689:Hodgkin Disease / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism; D017730:Ki-1 Antigen / Q000378:metabolism*; D013058:Mass Spectrometry; D018613:Microscopy, Confocal; D046529:Microscopy, Electron, Transmission; D016253:Microscopy, Immunoelectron; D053141:Organelle Size; D016539:Reed-Sternberg Cells / Q000276:immunology / Q000378:metabolism* / Q000648:ultrastructure; D022142:Secretory Vesicles / Q000276:immunology / Q000378:metabolism* / Q000648:ultrastructure; D015398:Signal Transduction; D059016:Tumor Microenvironment
chemical_list: D014408:Biomarkers, Tumor; D053285:CD30 Ligand; C508606:CXCL8 protein, human; D016209:Interleukin-8; D017730:Ki-1 Antigen; C505774:TNFSF8 protein, human
keywords: CD30; Hodgkin's lymphoma; TNF receptor superfamily; actin; extracellular vesicle; immune cell; shedding; tubulin; tumour microenvironment