Home LiteratureArticle Details
PMID: 15111327 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Proteomic analysis of exosomes secreted by human mesothelioma cells.

The American journal of pathology ·Vol. 164 ·No. 5 ·2004-05-00 ·Pages 1807-15

Hegmans JP, Bard MP, Hemmes A, Luider TM, Kleijmeer MJ, Prins JB, Zitvogel L, Burgers SA, Hoogsteden HC, Lambrecht BN

Abstract

Exosomes are small membrane vesicles secreted into the extracellular compartment by exocytosis. Tumor exosomes may be involved in the sampling of antigens to antigen presenting cells or as decoys allowing the tumor to escape immune-directed destruction. The proteins present in exosomes secreted by tumor cells have been poorly defined. This study describes the protein composition of mesothelioma cell-derived exosomes in more detail. After electrophoresis of exosome preparations, matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) was used to characterize the protein spots. MHC class I was found to be present together with the heat shock proteins HSC70 and HSP90. In addition, we found annexins and PV-1, proteins involved in membrane transport and function. Cytoskeleton proteins and their associated proteins ezrin, moesin, actinin-4, desmoplakin, and fascin were also detected. Besides the molecular motor kinesin-like protein, many enzymes were detected revealing the cytoplasmic orientation of exosomes. Most interesting was the detection of developmental endothelial locus-1 (DEL-1), which can act as a strong angiogenic factor and can increase the vascular development in the neighborhood of the tumor. In conclusion, mesothelioma cells release exosomes that express a discrete set of proteins involved in antigen presentation, signal transduction, migration, and adhesion. Exosomes may play an important role in the interaction between tumor cells and their environment.

MeSH Terms
Antigen-Presenting Cells/chemistry Blotting, Western Cell Adhesion Cell Line, Tumor Cell Movement Cytoplasm/metabolism Cytoskeleton/metabolism DNA/chemistry Exocytosis Exoribonucleases HSC70 Heat-Shock Proteins HSP70 Heat-Shock Proteins/chemistry HSP90 Heat-Shock Proteins/chemistry Histocompatibility Testing Humans Immunohistochemistry Karyotyping Lung Neoplasms/metabolism,pathology Mesothelioma/metabolism,pathology Microscopy, Electron Proteins/chemistry Proteome Signal Transduction Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Chemicals
HSC70 Heat-Shock Proteins HSP70 Heat-Shock Proteins HSP90 Heat-Shock Proteins HSPA8 protein, human Proteins Proteome DNA Exoribonucleases
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Hegmans Joost P J J
Department of Pulmonary Medicine, Erasmus MC, Rotterdam, The Netherlands. j.hegmans@erasmusmc.nl
Bard Martin P L
Hemmes Annabrita
Luider Theo M
Kleijmeer Monique J
Prins Jan-Bas
Zitvogel Laurence
Burgers Sjaak A
Hoogsteden Henk C
Lambrecht Bart N
References (62)
62 references, click to expand
  1. Proteomic and biochemical analyses of human B cell-derived exosomes. Potential implications for their function and multivesicular body formation.
    J Biol Chem. 2003 Mar 28;278(13):10963-72 PMID: 12519789
  2. On the origin of cancer cells.
    Science. 1956 Feb 24;123(3191):309-14 PMID: 13298683
  3. Eradication of established murine tumors using a novel cell-free vaccine: dendritic cell-derived exosomes.
    Nat Med. 1998 May;4(5):594-600 PMID: 9585234
  4. Molecular characterization of dendritic cell-derived exosomes. Selective accumulation of the heat shock protein hsc73.
    J Cell Biol. 1999 Nov 1;147(3):599-610 PMID: 10545503
  5. CD91: a receptor for heat shock protein gp96.
    Nat Immunol. 2000 Aug;1(2):151-5 PMID: 11248808
  6. Exosome: from internal vesicle of the multivesicular body to intercellular signaling device.
    J Cell Sci. 2000 Oct;113 Pt 19:3365-74 PMID: 10984428
  7. Desmoplakin is essential in epidermal sheet formation.
    Nat Cell Biol. 2001 Dec;3(12):1076-85 PMID: 11781569
  8. cDNA and protein sequence, genomic organization, and analysis of cis regulatory elements of mouse and human PLVAP genes.
    Genomics. 2001 Mar 15;72(3):304-13 PMID: 11401446
  9. Fascin, an actin-bundling protein associated with cell motility, is upregulated in hormone receptor negative breast cancer.
    Br J Cancer. 2000 Oct;83(7):870-3 PMID: 10970687
  10. Characteristics of the interaction between Hsc70 and the transferrin receptor in exosomes released during reticulocyte maturation.
    J Biol Chem. 2001 Mar 30;276(13):9910-6 PMID: 11133993
  11. The embryonic angiogenic factor Del1 accelerates tumor growth by enhancing vascular formation.
    Microvasc Res. 2002 Jul;64(1):148-61 PMID: 12074641
  12. Characterization of heat shock protein 110 and glucose-regulated protein 170 as cancer vaccines and the effect of fever-range hyperthermia on vaccine activity.
    J Immunol. 2001 Jan 1;166(1):490-7 PMID: 11123328
  13. Mutations in ACTN4, encoding alpha-actinin-4, cause familial focal segmental glomerulosclerosis.
    Nat Genet. 2000 Mar;24(3):251-6 PMID: 10700177
  14. Tumor-derived exosomes are a source of shared tumor rejection antigens for CTL cross-priming.
    Nat Med. 2001 Mar;7(3):297-303 PMID: 11231627
  15. Neovascularization of ischemic tissues by gene delivery of the extracellular matrix protein Del-1.
    J Clin Invest. 2003 Jul;112(1):30-41 PMID: 12840057
  16. Purification of heat shock protein-peptide complexes for use in vaccination against cancers and intracellular pathogens.
    Methods. 1997 Jun;12(2):165-71 PMID: 9184380
  17. ERM (ezrin/radixin/moesin) family: from cytoskeleton to signal transduction.
    Curr Opin Cell Biol. 1997 Feb;9(1):70-5 PMID: 9013673
  18. Cutting edge: receptor-mediated endocytosis of heat shock proteins by professional antigen-presenting cells.
    J Immunol. 1999 Apr 1;162(7):3757-60 PMID: 10201889
  19. Cloning and chromosomal localization of the human cytoskeletal alpha-actinin gene reveals linkage to the beta-spectrin gene.
    Am J Hum Genet. 1990 Jul;47(1):62-72 PMID: 2349951
  20. Annexins: from structure to function.
    Physiol Rev. 2002 Apr;82(2):331-71 PMID: 11917092
  21. C-erbB-2/ HER-2 upregulates fascin, an actin-bundling protein associated with cell motility, in human breast cancer cell lines.
    Oncogene. 2000 Oct 5;19(42):4864-75 PMID: 11039904
  22. A point mutation in the alpha-actinin-4 gene generates an antigenic peptide recognized by autologous cytolytic T lymphocytes on a human lung carcinoma.
    Cancer Res. 2001 May 15;61(10):4078-83 PMID: 11358829
  23. B lymphocytes secrete antigen-presenting vesicles.
    J Exp Med. 1996 Mar 1;183(3):1161-72 PMID: 8642258
  24. Malignant effusions and immunogenic tumour-derived exosomes.
    Lancet. 2002 Jul 27;360(9329):295-305 PMID: 12147373
  25. Stress-induced proteins in immune response to cancer.
    Curr Top Microbiol Immunol. 1991;167:109-23 PMID: 1711433
  26. Identity of p36K phosphorylated upon Rous sarcoma virus transformation with a protein purified from brush borders; calcium-dependent binding to non-erythroid spectrin and F-actin.
    EMBO J. 1984 Jan;3(1):227-33 PMID: 6323166
  27. Actinin-4, a novel actin-bundling protein associated with cell motility and cancer invasion.
    J Cell Biol. 1998 Mar 23;140(6):1383-93 PMID: 9508771
  28. Glucose metabolism in human gliomas: correspondence of in situ and in vitro metabolic rates and altered energy metabolism.
    Metab Brain Dis. 1986 Dec;1(4):279-91 PMID: 3508247
  29. Exosomes: composition, biogenesis and function.
    Nat Rev Immunol. 2002 Aug;2(8):569-79 PMID: 12154376
  30. Phylogenetic conservation of the molecular and immunological properties of the chaperones gp96 and hsp70.
    Eur J Immunol. 2001 Jan;31(1):186-95 PMID: 11265634
  31. Fascin, a sensitive new marker for Reed-Sternberg cells of hodgkin's disease. Evidence for a dendritic or B cell derivation?
    Am J Pathol. 1997 Feb;150(2):543-62 PMID: 9033270
  32. Comparison of tumor-specific immunogenicities of stress-induced proteins gp96, hsp90, and hsp70.
    J Immunol. 1994 Jun 1;152(11):5398-403 PMID: 8189059
  33. Annexin I is phosphorylated in the multivesicular body during the processing of the epidermal growth factor receptor.
    J Cell Biol. 1993 Jan;120(1):77-83 PMID: 8093248
  34. Protein folding in the cell.
    Nature. 1992 Jan 2;355(6355):33-45 PMID: 1731198
  35. Cloning and characterization of developmental endothelial locus-1: an embryonic endothelial cell protein that binds the alphavbeta3 integrin receptor.
    Genes Dev. 1998 Jan 1;12(1):21-33 PMID: 9420328
  36. Proteomic analysis of exosomes isolated from human malignant pleural effusions.
    Am J Respir Cell Mol Biol. 2004 Jul;31(1):114-21 PMID: 14975938
  37. Cancer vaccines: finding the best way to train the immune system.
    J Natl Cancer Inst. 2002 Oct 16;94(20):1523-6 PMID: 12381703
  38. Cytotoxic T lymphocytes directed against a tumor-specific mutated antigen display similar HLA tetramer binding but distinct functional avidity and tissue distribution.
    Proc Natl Acad Sci U S A. 2002 Jul 9;99(14):9358-63 PMID: 12093915
  39. Proteomic analysis of dendritic cell-derived exosomes: a secreted subcellular compartment distinct from apoptotic vesicles.
    J Immunol. 2001 Jun 15;166(12):7309-18 PMID: 11390481
  40. The annexins and exocytosis.
    Science. 1992 Nov 6;258(5084):924-31 PMID: 1439804
  41. RhoA-dependent phosphorylation and relocalization of ERM proteins into apical membrane/actin protrusions in fibroblasts.
    Mol Biol Cell. 1998 Feb;9(2):403-19 PMID: 9450964
  42. The cDNA sequence of a human placental alpha-actinin.
    Nucleic Acids Res. 1989 Aug 25;17(16):6725 PMID: 2780298
  43. Oncogenic alterations of metabolism.
    Trends Biochem Sci. 1999 Feb;24(2):68-72 PMID: 10098401
  44. 'The metabolism of tumours': 70 years later.
    Novartis Found Symp. 2001;240:251-60; discussion 260-4 PMID: 11727934
  45. Antitumor cytotoxic T-lymphocyte response in human lung carcinoma: identification of a tumor-associated antigen.
    Immunol Rev. 2002 Oct;188:114-21 PMID: 12445285
  46. PV-1 is a component of the fenestral and stomatal diaphragms in fenestrated endothelia.
    Proc Natl Acad Sci U S A. 1999 Nov 9;96(23):13203-7 PMID: 10557298
  47. Calreticulin, a peptide-binding chaperone of the endoplasmic reticulum, elicits tumor- and peptide-specific immunity.
    J Exp Med. 1999 Mar 1;189(5):797-802 PMID: 10049943
  48. Cellular tumorigenicity in nude mice: correlation with cell growth in semi-solid medium.
    Cell. 1974 Dec;3(4):355-9 PMID: 4442124
  49. The epithelial cell cytoskeleton and intracellular trafficking. II. Intestinal epithelial cell exosomes: perspectives on their structure and function.
    Am J Physiol Gastrointest Liver Physiol. 2002 Aug;283(2):G251-5 PMID: 12121870
  50. CD91 is a common receptor for heat shock proteins gp96, hsp90, hsp70, and calreticulin.
    Immunity. 2001 Mar;14(3):303-13 PMID: 11290339
  51. Anti-tumour immunotherapy using dendritic-cell-derived exosomes.
    Res Immunol. 1998 Sep-Oct;149(7-8):661-2 PMID: 9851520
  52. Cloning and characterization of two human skeletal muscle alpha-actinin genes located on chromosomes 1 and 11.
    J Biol Chem. 1992 May 5;267(13):9281-8 PMID: 1339456
  53. Annexins: the problem of assessing the biological role for a gene family of multifunctional calcium- and phospholipid-binding proteins.
    Biochim Biophys Acta. 1994 Apr 5;1197(1):63-93 PMID: 8155692
  54. In adrenocortical tissue, annexins II and VI are attached to clathrin coated vesicles in a calcium-independent manner.
    Biochim Biophys Acta. 1998 Mar 27;1402(2):115-30 PMID: 9561798
  55. ERM proteins: head-to-tail regulation of actin-plasma membrane interaction.
    Trends Biochem Sci. 1997 Feb;22(2):53-8 PMID: 9048483
  56. Annexin VI-mediated loss of spectrin during coated pit budding is coupled to delivery of LDL to lysosomes.
    J Cell Biol. 1998 Aug 24;142(4):937-47 PMID: 9722607
  57. Dendritic cells or their exosomes are effective biotherapies of cancer.
    Eur J Cancer. 1999 Aug;35 Suppl 3:S36-8 PMID: 10645221
  58. Heat shock proteins in health and disease: therapeutic targets or therapeutic agents?
    Expert Rev Mol Med. 2001 Sep 21;3(23):1-21 PMID: 14585147
  59. Malignant pleural mesothelioma.
    Crit Rev Oncol Hematol. 1997 Feb;25(2):97-126 PMID: 9134313
  60. Distribution and characterization of plasmalemma vesicle protein-1 in rat endocrine glands.
    J Endocrinol. 2002 Dec;175(3):649-61 PMID: 12475376
  61. Selective enrichment of tetraspan proteins on the internal vesicles of multivesicular endosomes and on exosomes secreted by human B-lymphocytes.
    J Biol Chem. 1998 Aug 7;273(32):20121-7 PMID: 9685355
  62. Molecular chaperones and the immune response.
    Philos Trans R Soc Lond B Biol Sci. 1993 Mar 29;339(1289):363-7; discussion 367-8 PMID: 7684139
Article Info
Journal
The American journal of pathology
Abbr.
Am J Pathol
ISSN
0002-9440
DOI
10.1016/S0002-9440(10)63739-X
Email
j.hegmans@erasmusmc.nl
Published
2004-05-00
Pages
1807-15
Language
English
Region
United States
NLM ID
0370502
PMCID
PMC1615654
Subset
IM
External Links
PubMed source DOI: 10.1016/S0002-9440(10)63739-X PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2