Tumour-derived exosomes and their role in cancer-associated T-cell signalling defects.
Dendritic and lymphoid 'exosomes' regulate immune activation. Tumours release membranous material mimicking these 'exosomes,' resulting in deletion of reactive lymphocytes. Tumour-derived 'exosomes' have recently been explored as vaccines, without analysis of their immunologic consequences. This investigation examines the composition of tumour-derived 'exosomes' and their effects on T lymphocytes. Membranous materials were isolated from ascites of ovarian cancer patients (n=6) and Western immunoblotting was performed for markers associated with 'exosomes.' Using cultured T cells, 'exosomes' were evaluated for suppression of CD3-zeta and JAK 3 expressions and induction of apoptosis, measured by DNA fragmentation. 'Exosome' components mediating suppression of CD3-zeta were isolated by continuous eluting electrophoresis and examined by Western immunoblotting. 'Exosomes' were shown to be identical with previously characterised shed membrane vesicles by protein staining and TSG101 expression. 'Exosomes' expressed class I MHC, placental alkaline phosphatase, B23/nucleophosmin, and FasL. 'Exosomes' suppressed expression of T-cell activation signalling components, CD3-zeta and JAK 3 and induced apoptosis. CD3-zeta suppression was mediated by two components: 26 and 42 kDa. Only the 42 kDa component reacted with anti-FasL antibody. These results indicate that, while 'exosomes' express tumour antigens, leading to their proposed utility as tumour vaccines, they also can suppress T-cell signalling molecules and induce apoptosis.
Taylor
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DD
Department of Obstetrics, Gynecology & Women's Health, University of Louisville School of Medicine, Louisville, KY 40202, USA. ddtaylor@louisville.edu
Gerçel-Taylor
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D000951:Antigens, Neoplasm / Q000032:analysis / Q000276:immunology; D015153:Blotting, Western; D017252:CD3 Complex / Q000096:biosynthesis; D002462:Cell Membrane / Q000737:chemistry / Q000276:immunology*; D002478:Cells, Cultured; D022162:Cytoplasmic Vesicles / Q000737:chemistry / Q000276:immunology*; D053938:DNA Fragmentation; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D015395:Histocompatibility Antigens Class I / Q000276:immunology; D006801:Humans; D053616:Janus Kinase 3; D008213:Lymphocyte Activation / Q000276:immunology; D010051:Ovarian Neoplasms / Q000276:immunology*; D011505:Protein-Tyrosine Kinases / Q000096:biosynthesis; D015398:Signal Transduction / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology*
D000951:Antigens, Neoplasm; D017252:CD3 Complex; D015395:Histocompatibility Antigens Class I; D011505:Protein-Tyrosine Kinases; C507933:JAK3 protein, human; D053616:Janus Kinase 3