Proteomic Analysis of Exosomes Isolated from Human Malignant Pleural Effusions
Abstract
Exosomes are membrane vesicles from endosomal origin secreted by various cells such as hematopoietic, epithelial, and tumor cells. Exosomes secreted by tumor cells contain specific antigens potentially useful for immunotherapeutic purposes. Our aim was to determine if exosomes are present in human cancerous pleural effusions and to identify their proteomic content. Exosomes were purified by sucrose gradient ultracentrifugation, and electron microscopy was used to check both concentration and purity of exosomes. Proteins were separated by one-dimensional sodium dodecyl sulfate–polyacrylamide gel electrophoresis, and protein bands were identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry and Western blotting. Exosomes were present in pleural fluid obtained from patients suffering from mesothelioma (n = 4), lung cancer (n = 2), breast cancer (n = 2), and ovarian cancer (n = 1). As previously reported by others, antigen-presenting molecules, cytoskeletal proteins, and signal transduction–involved proteins were present. Proteins not previously reported were identified (SNX25, BTG1, PEDF, thrombospondin 2). Different types of immunoglobulins and complement factors were abundantly present in the sucrose fractions containing exosomes. Exosome-directed specificity of these immunoglobulins was not observed. In conclusion, sucrose gradient ultracentrifugation allows isolation of exosomes from malignant pleural effusions. However, pleural fluid proteins and especially immunoglobulins are coisolated and may hamper the use of exosomes isolated from malignant effusion for immunotherapy programs.
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CD63 and CD9 are potential exosome markers; ARF6 and VAMP3 are proposed microvesicle markers; TSP and C3b are accepted apoptotic body markers; Gag protein may be an RLP marker.
“CD63 and CD9 are potential markers of exosomes [22, 117]. Markers of microvesicles are less well established though ARF6 and VCAMP3 are recently proposed [66]. TSP and C3b are generally accepted markers of apoptotic bodies. The RLPs are less well studied though Gag protein may be a marker for this group of EVs (Table 1).”
From Biogenesis of extracellular vesicles (EV): exosomes, microvesicles, retrovirus-like vesicles, and apoptotic bodies (2013) · cites this paper as [117]
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BackgroundThe Immune Microenvironment of Malignant Pleural Mesothelioma: A Literature Review
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- Authors
- Bard, M. P., Hegmans, J. P., Hemmes, A., Luider, T. M., Willemsen, R., Severijnen, L.-A. A., van Meerbeeck, J. P., Burgers, S. A., Hoogsteden, H. C., & Lambrecht, B. N.
- Affiliations
- Erasmus MC (NL)
- Department of Pulmonary Medicine, Department of Neurology, and Department of Clinical Genetics, Erasmus MC, Rotterdam, The Netherlands
- ORCID
- Venue
- American Journal of Respiratory Cell and Molecular Biology
- Publisher
- Oxford University Press (OUP)
- Published
- 2004-07-01
- Volume
- 31
- Issue
- 1
- Pages
- 114-121
- ISSN
- 1044-1549 (ISSN-L)
- 1535-4989
- OpenAlex
- W2089081493
- Cited by
- 416
- References
- 2
- Citations by year
- 2026: 5 · 2025: 13 · 2024: 15 · 2023: 19 · 2022: 17 · 2021: 18 · 2020: 28 · 2019: 15 · 2018: 25 · 2017: 25 · 2016: 23 · 2015: 32 · 2014: 26 · 2013: 25 · 2012: 16
- FWCI
- 4.62
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- American Journal of Respiratory Cell and Molecular Biology (publishedVersion)
- PubMed (publishedVersion)
- EUR Research Repository (Erasmus University Rotterdam) (submittedVersion)
- Ghent University Academic Bibliography (Ghent University) (submittedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
- Health Sciences › Medicine › Pulmonary and Respiratory Medicine › Pleural and Pulmonary Diseases
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Keywords
- exosomes, malignant pleural effusion, exosomal proteome, mesothelioma, lung cancer, breast cancer, ovarian cancer, immunoglobulins, complement proteins, antigen-presenting molecule, BTG1, pigment epithelium-derived factor, thrombospondin-2, exosome isolation
- MeSH terms
- Pigment Epithelium-Derived Factor, Thrombospondin 2, Aged, Breast Neoplasms, Carcinoma, Carrier Proteins, Complement System Proteins, Eye Proteins, Female, Humans, Immunoglobulins, Lung Neoplasms, Male, Mesothelioma, Microscopy, Electron, Middle Aged, Neoplasm Proteins, Nerve Growth Factors, Ovarian Neoplasms, Predictive Value of Tests, Proteins, Endosomes, Biomarkers, Tumor, Ultracentrifugation, Serpins, Pleural Effusion, Malignant, Thrombospondins, Secretory Vesicles, Vesicular Transport Proteins, Proteomics, Sorting Nexins
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