Chinese Bulletin of Life Sciences. The roles of exosomes in immune tolerance. ZHAO Lihua 1,2, FAN Huahua 2 *

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19 2 2007 4 Chinese Bulletin of Life Sciences Vol. 19, No. 2 Apr., 2007 1004-0374(2007)02-0174-05 exosome (1, 200062 2, 200051), exosome exosome exosome exosome exosome R392A The roles of exosomes in immune tolerance ZHAO Lihua 1,2, FAN Huahua 2 * (1 Department of Biomedical Science, School of Life Sciences, East China Normal University, Shanghai 200062, China; 2 Laboratory for Blood Engineering, Shanghai (Red cross) Blood Center, Shanghai 200051, China) Abstract: Exosomes are small membrane vesicles of endocytic origin that are secreted by many live cells. The specific functions of the different exosomes are correlated closely with the cell-specific proteins and the microenvironment of exosomes. Previous researches focus on the roles of exosome in stimulating and intensifying immunity. Recently, more and more studies show that exosomes can also down-regulate immune response or induce immune tolerance in specific environments. And especially more and more researchers pay attention to the roles of immune tolerance induced by exosomes in allotransplantation and autoimmune diseases. Therefore the applications for many diseases therapy through immune tolerance by exosomes will become a research hotspot. In this review we focus on exosomes biogenesis and the roles of induction of immune tolerance. Key words: exosomes; immunes tolerance; allotransplantation; autoimmune diseases; tumor immunity [1] exosome exosome (tumor-cell-derived exosome TEXs) DC exosome (dendritic-cell-derived exosome DEXs) exosome exosome 2006-08-14 2006-11-21 (0552nm010) (1982 )(1966 ) * Tel: 021-62957482 E-mail: fhh021@hotmail.com

2 exosome 175 exosome exosome 1 exosome exosome [1-3] [3] exosome (multivesicular bodys MVBs) [1] MVBs MVBs [1, 3] MVBs MVBs MVBs MVBs MHC-II MHC-II MVBs exosome MVBs [2-3] exosome MVBs Ca 2+ Stoorvogel [3] K562 exosomeca 2+ Clayton [4] Ca 2+ A23187 Burkil exosome exosome 2 exosome 2.1 exosome exosome 30 100 nm [1, 5] exosome 1.13 1.21 g/ml [1, 5] B exosome 60 80nm MVBs MVBs exosome [6] 2.2 exosome exosome Western blotting FACS exosome [1-3, 5-6] exosome exosome [5-6] ( ) (annexin rab ) ( G ) ( - 1) (Hsp70 Hsp84 Hsp90 ) exosome tetraspanins (CD9 CD63 CD81 CD82) [1,6] MHC exosome APC exosome [1] DEXs B exosome(bexs) MHC- II (CD86 CD80) T TEXs exosome LMP1 [5] T TEXs FasL AICD HLA-G [7] DEXs MFG-E8 [1,6,8] exosome DC DC T exosome A33 exosome P exosome N/CD13 [1,6] exosome BEXs

176 19 DEXs [6] 3 exosome exosome exosome [2] APC exosome MHC APC exosome DC exosome [1] exosome exosome exosome 3.1 (IECs)exosome Karlsson [9] exosome exosome Mallegol [10] IFN-γ IECs MODEK (OVA) IFN-γ OVA OVA IFN-γ exosome OVA IFN-γ exosome OVA Ostman [11] exosome( ) MHC 1983 Strobel [12] OVA 1h (DTH) OVA OVA IECs IECs 40 nm exosome MHC OVA OVA MHC IECs [9,11] SCID IECs MHC SCID IFN-γ IECs IFN-γ SCID IECs MHC IFN-γ MHC MHC [11] IECs MHC CD4 + T (Treg ) TGF-β IL-10 [5,9] Th1 CD4 + T 3.2 DC exosome(dexs) DC APC DC DC T [13] DEXs [14] DEXs Peche [15] BMDC DEXs( RT1u) DEXs(RT1a) 1 RT1u IFN-γ mrna 7 d 14 d 2 RT1u(1 10 25 100 µg) 10 µg DEXs [8] BMDC exosome LF15-0195 LF15-0195 DC DEXs

2 exosome 177 [16] Morelli [17] exosome exosomedc Kupffer DC exosome DC exosome CD8α DC CD8α + T MHC exosome CD4 + T exosome DC Kim [18] FasL DC DCFasL FasL DC exosome IL-10 BMDC IL-10 BMDC exosome [19] 3.3 exosome (TEXs) TEXs (TAA) APC T [20] TEXs CTL TEXs T [21-24] TEXs T Abusamra [21] FasL TEXs T TEXs TEXs FasL Fas T Fas Fas/FasL AICD T TEXs FasL T T [5] TEXs FasL [21] TEXs TEXs HLA-G [7] HLA-G NK CTL [5, 7] HLA-G exosome HLA-G TEXs TEXs EMP1 T Taylor Gercel-Talor [22] TEXs T CD3- JAK 3 T TEXs 3.4 exosome Frangsmyr [23] ( ) FasL exosome FasL exosome Fas [5] Waterhouse [24] exosome 17 CD9 Gansuvd [25] EB B exosome LMP1 CD4 + T (Treg ) T T exosome [2] 4 exosome

178 19 exosome exosome exosome exosome exosome [1] Thery C, Zitvoge L, Amigorena S. Exosome: composition, biogenesis and function. Nat Rev Immunol, 2002, 2 (8): 569-579 [2]. Exosome.., 2004, 4(31) : 185-188 [3] Stoorvogel W, Kleijmeer M J, Geuze H J, et al. The biogenesis and functions of exosome. Traffic, 2002, 3 (5): 321-330 [4] Clayton A, Court J, Navabi H, et al. Analysis of antigen presenting cell derived exosome, based on immuno-magnetic isolation and flow cytometry. J Immunol Methods, 2001, 247(1-2): 163-174 [5] Xiu F M, Cao X T. Exosome in the immune response and tolerance. J Microbiol Immunol, 2004, 2 (4): 231-235 [6] Chaput N, Taïeb J, Schartz N E C, et al. Exosome-based immunotherapy. Cancer Immunol Immunother, 2004, 53 (3): 234-239 [7] Riteau B, Faure F, Menier C, et al. Exosome bearing HLA-G are released by melanoma cells. Hum Immunol, 2003, 64 (11): 1064-1072 [8] Oshima K, Aoki N, Kato T, et al. Secretion of a peripheral membrane protein, MFG-E8, as a complex with membrane vesicles. Eur J Biochem, 2002, 269 (4): 1209-1218 [9] Karlsson M, Lundin S, Dahlgren U, et al. Tolerosomes are produced by intestinal epithelial cells. Eur J Immunol, 2001, 31(10): 2892-2900 [10] Mallegol J, Van Niel G, Heyman M. Phenotypic and functional characterization of intestinal epithelial exosome. Blood Cells Mol Dis, 2005, 35 (1): 11-16 [11] Ostman S, Taube M, Telemo E. Tolerosome-induced oral tolerance is MHC dependent. Immunology, 2005, 116 (4): 464-476 [12] Strobel S, Mowat A M, Drummond H E, et al. Immunological responses to fed protrin antigens in mice, II, Oral tolerance for CMI is due to activation of cyclophosphamidesensitive cells by gut-processed antigen. Immunology, 1983, 49 (3): 451-456 [13] Ichim T E, Zhong R, Min W P. Prevention of allograft rejection by in vitro generated tolerogenic dendritic cell. Transpl Immunol, 2003, 11 (3-4): 295-306 [14] Chaput N, Julien Taieb J, Schartz N, et al. The potential of exosome in immunotherapy of cancer. Blood Cells Mol Dis, 2005, 35 (2): 111-115 [15] Peche H, Heslan M, Usal C, et al. Presentation of donor major hidtocompatibility complex antigens by bone marrow dendritic cell derived exosome modulates allograft rejection 1. Transplantation, 2003, 76(10): 1503-1510 [16] Peche H, Renaudin K, Beriou G, et al. Induction of tolerance by exosome and short-term immunosuppression in a fully MHC-mismatched rat cardiac allograft model. Am J Transplantat, 2006, 6 (7): 1541-1550 [17] Morelli A E, Larregina A T, Shufesky W J, et al. Endocytosis intracellular sorting, and processing of exosome by dendritic cells. Blood, 2004, 104 (10): 3257-3266 [18] Kim S H, Bianco N, Menon R, et al. Exosome derived from genetically modified DC expressing FasL are anti-inflammatory and immunosuppressive. Mol Ther, 2006, 13(2): 289-300 [19] Kim S H, Lechman E R, Bianco N, et al. Exosome derived from IL-10-treated dendritic cells can suppress inflammation and collagen-induced arthritis. J Immunol, 2005, 174 (10): 6440-6448 [20] Wolfers J, Lozier A, Raposo G, et al. Tumor-derived exosomes are a source of shared tumor rejection antigens for CTL cross-priming. Nat Med, 2001, 7 (3): 297-303 [21] Abusamra A J, Zhong Z H, Zheng X F, et al. Tumor exosome expressing Fas ligand mediate CD8 + T-cell apoptosis. Blood Cells Mol Dis, 2005, 35 (2): 169-173 [22] Taylor D D, Gercel-Taylor C. Tumour-derived exosome and their role in cancer-associated T-cell signalling defects. Br J Cancer, 2005, 92(2):305-311 [23] Frangsmyr L, Baranov V, Nagaeva O, et al. Cytoplasmic microvesicular form of Fas ligand in human early placenta: switching the tissue immune privilege hypothesis from cellular to vesicular level. Mol Hum Reprod, 2005, 11(1): 35-41 [24] Waterhouse R, Ha C, Dveksler G S. Murine CD9 is the recepter for pregnancy-specific glycoprotein 17. J Exp Med, 2002, 195 (2): 277-282 [25] Gansuvd B, Hagihara M, Higuchi A, et al.umbilical cord blood dendritic cells are a rich source of soluble HLA-DR: synergistic effedt of exosomes and dendritic cells on autologous or allogeneic T-Cell proliferation.hum Immunol, 2003, 64 (4): 427-439