Exosome: from internal vesicle of the multivesicular body to intercellular signaling device

2000 ◽  
Vol 113 (19) ◽  
pp. 3365-3374 ◽  
Author(s):  
K. Denzer ◽  
M.J. Kleijmeer ◽  
H.F. Heijnen ◽  
W. Stoorvogel ◽  
H.J. Geuze
Keyword(s):  
Dendritic Cells ◽  
B Lymphocyte ◽  
Cytotoxic T Cells ◽  
Membrane Vesicles ◽  
Multivesicular Body ◽  
Cell Types ◽  
Target Cells ◽  
Intercellular Signaling ◽  
Accessory Cells

Exosomes are small membrane vesicles that are secreted by a multitude of cell types as a consequence of fusion of multivesicular late endosomes/lysosomes with the plasma membrane. Depending on their origin, exosomes can play roles in different physiological processes. Maturing reticulocytes externalize obsolete membrane proteins such as the transferrin receptor by means of exosomes, whereas activated platelets release exosomes whose function is not yet known. Exosomes are also secreted by cytotoxic T cells, and these might ensure specific and efficient targeting of cytolytic substances to target cells. Antigen presenting cells, such as B lymphocytes and dendritic cells, secrete MHC class-I- and class-II-carrying exosomes that stimulate T cell proliferation in vitro. In addition, dendritic-cell-derived exosomes, when used as a cell-free vaccine, can eradicate established murine tumors. Although the precise physiological target(s) and functions of exosomes remain largely to be resolved, follicular dendritic cells (accessory cells in the germinal centers of secondary lymphoid organs) have recently been shown to bind B-lymphocyte-derived exosomes at their cell surface, which supports the notion that exosomes play an immunoregulatory role. Finally, since exosomes are derived from multivesicular bodies, their molecular composition might provide clues to the mechanism of protein and lipid sorting in endosomes.

scholarly journals Surface membrane vesicles from mononuclear cells stimulate erythroid stem cells to proliferate in culture

Blood ◽  
1982 ◽  
Vol 60 (3) ◽  
pp. 583-594 ◽  
Author(s):  
N Dainiak ◽  
CM Cohen
Keyword(s):  
Plasma Membrane ◽  
Plasma Membranes ◽  
Mononuclear Cells ◽  
Surface Membrane ◽  
Freeze Fracture ◽  
Membrane Vesicles ◽  
Cell Types ◽  
Target Cells ◽  
Freeze Fracture Electron Microscopy

Abstract In order to examine the contribution of cell surface materials to erythroid burst-promoting activity (BPA), we separated media conditioned by a variety of human cell types into pellets and supernatants by centrifugation. When added to serum-restricted cultures of nonadherent human marrow cells, pellets contained about half of the total stimulatory activity. Freeze-fracture electron microscopy of the pellets revealed the presence of unilamellar membrane vesicles ranging from 0.10 to 0.40 microM in diameter. The amount of BPA in culture increased with added vesicle concentration in a saturable fashion. Preparation of leukocyte conditioned medium (LCM) from 125I-wheat germ agglutinin labeled cells and studies comparing the glycoprotein composition of vesicles with that of leukocyte plasma membranes suggest that LCM-derived vesicles are of plasma membrane origin. Moreover, partially purified leukocyte plasma membrane preparations also contained BPA. While disruption of vesicles by freezing/thawing and hypotonic lysis did not alter BPA, heat, trypsin, or pronase treatment removed greater than 65% of BPA, implying that vesicle surface rather than intravesicular molecules express BPA. Results of BPA assays performed in two-layer clots indicated that proximity to target cells is required for vesicle BPA expression. We conclude that membrane vesicles spontaneously shed from cell surfaces may be important regulators of erythroid burst proliferation in vitro.

Monosodium Urate Crystals Bind with Idiotype Protein and Promote Dendritic Cells to Induce Cytotoxic T Cells in Vitro

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 4904-4904
Author(s):  
Ippei Sakamaki ◽  
Kunihiro Inai ◽  
Takanori Ueda ◽  
Hiroshi Tsutani
Keyword(s):  
T Cells ◽  
Dendritic Cells ◽  
Cytotoxic T Cells ◽  
Target Cells ◽  
Monosodium Urate ◽  
Cd4 Cells ◽  
Antigenic Protein ◽  
Positively Charged ◽  
Msu Crystals

Abstract Monosodium urate (MSU) crystals have been studied to act as a key substance in local immunoreactions. MSU released from damaged cells works as an endogenous danger signal to antigen-presenting cells. MSU crystals evoke specific cell immunity and work as an adjuvant in a mouse model. The crystals also have another unique characteristic to bind with positively charged proteins, which could help to deliver some antigens into human dendritic cells (DCs). We focused on the application of MSU crystals as a not only an adjuvant but also as a carrier of positively charged antigenic protein to induce human cytotoxic T cells (CTLs) efficiently in vitro. We confirmed that MSU crystals facilitated human DCs to express the maturation marker, CD83, deliver (Fab′)2 attaching to the crystals. In order to determine whether MSU crystals facilitate the T-cell proliferation activity of DCs, the proliferative effects of DCs on allogeneic CD4+ cells were investigated. DCs pulsed with MSU crystals significantly facilitated the proliferation of allogeneic CD4+ cells when compared to DCs alone. The stimulation index (SI) was 2.5 ± 0.1 and 1.7 ± 0.1, respectively. When using DCs pulsed with the Fab attached to MSU crystals, the proliferation of CD4+ cells was significantly greater than when using DCs pulsed with Fab alone. The SI was 2.6 ± 0.2 and 1.9 ± 0.1, respectively. No significant differences were seen in the proliferation of allogeneic CD4+ cells between DCs pulsed with the Fab attached to MSU and DCs pulsed with MSU alone. We selected the multiple myeloma IM-9 cell line and its product idiotype (Id) protein as an ideal pair of target cells and positively charged tumor-specific antigen, respectively. After sensitizing DCs derived from HLA-A matched volunteers pulsed with tumor-specific monoclonal IgG-Fab fragments (IM-9 Fab) attached to MSU crystals, the CD8+ T cells stimulated by the DCs killed significantly more target cells (38.5 ± 3.5%, n=4) than those stimulated by DCs pulsed with IM-9 Fab alone (3.5 ± 7.5%). These cytotoxic effects of CD8+ cells stimulated by the DCs pulsed with IM-9 Fab attached to MSU crystals were reduced (3.6 ± 1.7%) when MSU crystals were pre-coated with fetal bovine serum to block to bind with IM-9 Fab. For efficient induction of CTLs, it is necessary for Id proteins to attach to MSU crystals. MSU crystals have some advantages of a protein carrier binding with positively charged proteins and delivering antigenic protein into DCs, as well as an adjuvant promoting DC maturation and inducing CTLs.

Induction of CML28-Specific Cytotoxic T Cells by Dendritic Cells Cotransfected with CML28 Nucleic Acid Vaccination and SOCS1-Specific siRNA Expression Vector In Vitro.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3058-3058
Author(s):  
Donghua Zhang ◽  
Hongsheng Zhou ◽  
Lu Zhang ◽  
Yaya Wang ◽  
Min Dai ◽  
...  
Keyword(s):  
T Cells ◽  
Dendritic Cells ◽  
Nucleic Acid ◽  
Gene Silencing ◽  
Expression Vector ◽  
Cytotoxic T Cells ◽  
Target Cells ◽  
Sirna Expression ◽  
Sirna Expression Vector

Abstract CML28 is a new tumor-associated antigen overexpressed in tumor cells, which is an attractive target for antigen-specific immunotherapy. In this study, we evaluated the possibility to induce CML28-specific cytotoxic T cells by cotransfection of CML28 nucleic acid vaccination and SOCS1-specific siRNA expression vector with dendritic cells in vitro. The full length CML28 cDNA was amplified from K562 by RT-PCR and cloned into a bicistronic vector pIRES2-EGFP to construct the CML28 nucleic acid vaccination. Dendritic cells (DCs) were generated from peripheral blood mononuclear cells (PBMNCs) of HLA-A2+ healthy donor. Construct the recombinant plasmid psiRNA-hH1neo-SOCS1 encoding hairpin small interfering RNA (siRNA) targeting to suppressors of cytokine signaling 1 (SOCS1) sequences using a vector-base RNA interference technology. Cotransfect CML28 nucleic acid vaccination and recombinant siRNA vector psiRNA-hH1neo-SOCS1 into DCs by electroporation. Real-time RT-PCR was performed to validate the SCOS1 gene silencing efficacy. The expression products of CML28 nucleic acid vaccination, His-CML28 fusion protein and GFP protein, were measured by Western Blotting and fluorescence microscope respectively. Labeling autologous nonadherent fraction of PBMNCs as responder cells by 5, 6-carboxyfluorescein diacetate succinimidyl ester (CFSE) firstly, and then coculture with cotransfected DCs for mixed lymphocyte reaction (MLR). Autogous T cell proliferative response of MLR was measured by FACS by detecting CFSE. Cotransfected DCs and the PBMNCs of a HLA-A2+ primary leukemia patient were used as target cells, untransfected DCs, K562 and HL60 were used as control. The standard 51Cr-release assay was performed to measure cytotoxicity of stimulated lyphocytes. CML28 nucleic acid vaccination could encode His-CML28 and GFP protein in DCs successfully. SOCS1-specific siRNA expression vector psiRNA-hH1neo-SOCS1 significantly suppress the expression of SOCS1 in DCs. Down regulation of SOCS1 resulted in higher expression level of CD80, CD86 and CD83 in cotranfected DCs and more rounds of cell division of responder cells in CFSE-MLR, which indicates SOCS1 gene silencing greatly contribute to maturation of DCs and enhance the the proliferative response of responder cells. CTLs induced by cotransfected DCs exhibit stronger CML28-specific cytotoxicity against HLA- matched target cells comparing to CTLs induced by CML28 nucleic acid vaccination transfected DCs only. SOCS1 gene silencing in DCs could greatly enhance the anti-tumor efficacy of CML28 nucleic acid vaccination. DCs cotransfected with CML28 nucleic acid vaccination and SOCS1-specific siRNA expression vector could effectively induce autologous CML28-specific cytotoxic T cells that lyse CML28 positive tumor cells in an antigen-specific and major histocompatibility complex (MHC) class I-restricted manner.

Dendritic Cells Tranfected with CML-28 cDNA Induce CML28-Specific Cytotoxic T Cells.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 5248-5248
Author(s):  
Hongsheng Zhou ◽  
Donghua Zhang ◽  
Yaya Wang ◽  
Yi Xiao ◽  
Wei Huang ◽  
...  
Keyword(s):  
Immune Response ◽  
Dendritic Cells ◽  
Tumor Antigen ◽  
Mononuclear Cells ◽  
Cytotoxic T Cells ◽  
Lymphocytic Leukemia ◽  
Myelogenous Leukemia ◽  
Target Cells ◽  
Peripheral Blood Mononuclear

Abstract Dendritic cells (DC) are the most potent antigen-presenting cells known; the use of DC loaded with tumor antigen is one of the most promising approaches to induce specific immune response. CML28 is an attractive candidate. According to the published article and our following study, CML28 is a broadly immunogenic antigen, which is overexpessed in leukemia cells, including acute myeloid leukemia (AML), acute lymphocytic leukemia (ALL) and chronic myelogenous leukemia (CML); but not in normal hematopoietic tissues or other normal tissue, with the exception of testis. In our study, DCs were generated from peripheral blood mononuclear cells (PBMC) by culture with GM-CSF, IL-4 and TNF-α. The full CML28 cDNA was amplified from K562 cell and then cloned into plasmid pcDNA3.1 HisA. Load DC with the constructed plasmid pcDNA3.1 HisA-CML28 by electroporation. The results of FACS analysis and Western Blot respectively demonstrate that the plasmid transfected DC retain immunophenotypical characteristics and can express the fusion protein CML28-His. Cytotoxic T lymphocytes (CTL) were generated from autologous PBMC stimulated by the transfected DCs. Cytoxicity assay in vitro revealed that CTL can recognize and lyse CML28-expressing target cells, such as the transfected DCs but not normal autologous DCs, which show that the plasmid pcDNA 3.1 HisA-CML28 transfected DC can induce CML28-specific CTL. Our studies demonstrate that CML28 can be a promising target of tumor antigen for leukemia-specific immunotherapy and DC loaded with CML28 tumor antigen can be a promising tool to induce specific anti-leukemia immune response.

scholarly journals Surface membrane vesicles from mononuclear cells stimulate erythroid stem cells to proliferate in culture

Blood ◽  
1982 ◽  
Vol 60 (3) ◽  
pp. 583-594
Author(s):  
N Dainiak ◽  
CM Cohen
Keyword(s):  
Plasma Membrane ◽  
Plasma Membranes ◽  
Mononuclear Cells ◽  
Surface Membrane ◽  
Freeze Fracture ◽  
Membrane Vesicles ◽  
Cell Types ◽  
Target Cells ◽  
Freeze Fracture Electron Microscopy

In order to examine the contribution of cell surface materials to erythroid burst-promoting activity (BPA), we separated media conditioned by a variety of human cell types into pellets and supernatants by centrifugation. When added to serum-restricted cultures of nonadherent human marrow cells, pellets contained about half of the total stimulatory activity. Freeze-fracture electron microscopy of the pellets revealed the presence of unilamellar membrane vesicles ranging from 0.10 to 0.40 microM in diameter. The amount of BPA in culture increased with added vesicle concentration in a saturable fashion. Preparation of leukocyte conditioned medium (LCM) from 125I-wheat germ agglutinin labeled cells and studies comparing the glycoprotein composition of vesicles with that of leukocyte plasma membranes suggest that LCM-derived vesicles are of plasma membrane origin. Moreover, partially purified leukocyte plasma membrane preparations also contained BPA. While disruption of vesicles by freezing/thawing and hypotonic lysis did not alter BPA, heat, trypsin, or pronase treatment removed greater than 65% of BPA, implying that vesicle surface rather than intravesicular molecules express BPA. Results of BPA assays performed in two-layer clots indicated that proximity to target cells is required for vesicle BPA expression. We conclude that membrane vesicles spontaneously shed from cell surfaces may be important regulators of erythroid burst proliferation in vitro.

scholarly journals Human Dendritic Cells Mediate Cellular Apoptosis via Tumor Necrosis Factor–Related Apoptosis-Inducing Ligand (Trail)

1999 ◽  
Vol 190 (8) ◽  
pp. 1155-1164 ◽  
Author(s):  
Neil A. Fanger ◽  
Charles R. Maliszewski ◽  
Ken Schooley ◽  
Thomas S. Griffith
Keyword(s):  
Dendritic Cells ◽  
Tumor Cells ◽  
Cell Types ◽  
Target Cells ◽  
Functional Difference ◽  
Cellular Apoptosis ◽  
Soluble Trail ◽  
Innate Effector ◽  
Human Dendritic Cells ◽  
Necrosis Factor

TRAIL (TNF-related apoptosis-inducing ligand) is a member of the TNF family that induces apoptosis in a variety of cancer cells. In this study, we demonstrate that human CD11c+ blood dendritic cells (DCs) express TRAIL after stimulation with either interferon (IFN)-γ or -α and acquire the ability to kill TRAIL-sensitive tumor cell targets but not TRAIL-resistant tumor cells or normal cell types. The DC-mediated apoptosis was TRAIL specific, as soluble TRAIL receptor blocked target cell death. Moreover, IFN-stimulated interleukin (IL)-3 receptor (R)α+ blood precursor (pre-)DCs displayed minimal cytotoxicity toward the same target cells, demonstrating a clear functional difference between the CD11c+ DC and IL-3Rα+ pre-DC subsets. These results indicate that TRAIL may serve as an innate effector molecule on CD11c+ DCs for the elimination of spontaneously arising tumor cells and suggest a means by which TRAIL-expressing DCs may regulate or eliminate T cells responding to antigen presented by the DCs.

scholarly journals Participation of the H-2 antigens of tumor cells in their lysis by syngeneic T cells.

1976 ◽  
Vol 143 (3) ◽  
pp. 601-614 ◽  
Author(s):  
J W Schrader ◽  
G M Edelman
Keyword(s):  
T Cells ◽  
Tumor Cell ◽  
Tumor Cells ◽  
Target Cell ◽  
Cytotoxic T Cells ◽  
Hybrid Origin ◽  
Target Cells ◽  
Cytotoxic Lymphocytes ◽  
Effector Cells

Cytotoxic T lymphocytes were generated in vitro against H-2 compatible or syngeneic tumor cells. In vitro cytotoxic activity was inhibited by specific anti-H2 sera, suggesting that H-2 antigens are involved in cell lysis. Two observations directly demonstrated the participation of the H-2 antigens on the tumor cells in their lysis by H-2-compatible T cells. First, coating of the H-2 antigens on the target tumor cell reduced the number of cells lysed on subsequent exposure to cytotoxic T cells. Second, when cytotoxic T cells were activated against an H-2 compatible tumor and assayed against an H-2-incompatible tumor, anti-H-2 serum that could bind to the target cell, but not to the cytotoxic lymphocyte, inhibited lysis. H-2 antigens were also shown to be present on the cytotoxic lymphocytes. Specific antisera reacting with these H-2 antigens, but not those of the target cell, failed to inhibit lysis when small numbers of effector cells were assayed against H-2-incompatible target cells or when effector cells of F1-hybrid origin and bearing two H-2 haplotypes were assayed against a tumor cell of one of the parental strains. These findings suggest that it is the H-2 antigens on the tumor cell and not those on the cytotoxic lymphocytes that are important in cell-mediated lysis of H-2-compatible tumor cells.

scholarly journals B cell survival, surface BCR and BAFFR expression, CD74 metabolism, and CD8− dendritic cells require the intramembrane endopeptidase SPPL2A

2012 ◽  
Vol 210 (1) ◽  
pp. 31-40 ◽  
Author(s):  
Hannes Bergmann ◽  
Mehmet Yabas ◽  
Alanna Short ◽  
Lisa Miosge ◽  
Nadine Barthel ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
B Cell ◽  
B Lymphocyte ◽  
B Cell Lymphoma ◽  
Cell Types ◽  
Proteolytic Processing ◽  
Mhc Ii ◽  
Humoral Immunodeficiency ◽  
B Cell Subsets ◽  
New Treatments

Druggable proteins required for B lymphocyte survival and immune responses are an emerging source of new treatments for autoimmunity and lymphoid malignancy. In this study, we show that mice with an inactivating mutation in the intramembrane protease signal peptide peptidase–like 2A (SPPL2A) unexpectedly exhibit profound humoral immunodeficiency and lack mature B cell subsets, mirroring deficiency of the cytokine B cell–activating factor (BAFF). Accumulation of Sppl2a-deficient B cells was rescued by overexpression of the BAFF-induced survival protein B cell lymphoma 2 (BCL2) but not BAFF and was distinguished by low surface BAFF receptor and IgM and IgD B cell receptors. CD8-negative dendritic cells were also greatly decreased. SPPL2A deficiency blocked the proteolytic processing of CD74 MHC II invariant chain in both cell types, causing dramatic build-up of the p8 product of Cathepsin S and interfering with earlier steps in CD74 endosomal retention and processing. The findings illuminate an important role for the final step in the CD74–MHC II pathway and a new target for protease inhibitor treatment of B cell diseases.

scholarly journals Differential Infectivity and Division ofToxoplasma gondii in Human Peripheral Blood Leukocytes

2000 ◽  
Vol 68 (8) ◽  
pp. 4822-4826 ◽  
Author(s):  
Jacqueline Y. Channon ◽  
Rosanne M. Seguin ◽  
Lloyd H. Kasper
Keyword(s):  
Dendritic Cells ◽  
Peripheral Blood ◽  
Human Peripheral Blood ◽  
Cell Types ◽  
Peripheral Blood Leukocytes ◽  
Blood Leukocytes ◽  
Differential Infectivity

ABSTRACT When tachyzoites were incubated with human peripheral blood leukocytes in vitro, more monocytes and dendritic cells than neutrophils or lymphocytes were infected. Although tachyzoites were able to divide in each of these cell types, monocytes and dendritic cells were more permissive to rapid tachyzoite division than neutrophils or lymphocytes.

scholarly journals Syncytin 1 dependent horizontal transfer of marker genes from retrovirally transduced cells

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Berna Uygur ◽  
Kamran Melikov ◽  
Anush Arakelyan ◽  
Leonid B. Margolis ◽  
Leonid V. Chernomordik
Keyword(s):  
Cell Lines ◽  
Membrane Vesicles ◽  
Retroviral Vectors ◽  
Extracellular Vesicle ◽  
Marker Genes ◽  
Target Cells ◽  
Retroviral Transduction ◽  
Gfp Gene ◽  
Safety Test

AbstractRetroviral transduction is routinely used to generate cell lines expressing exogenous non-viral genes. Here, we show that human cells transduced to stably express GFP transfer GFP gene to non-transduced cells. This horizontal gene transfer was mediated by a fraction of extracellular membrane vesicles that were released by the transduced cells. These vesicles carried endogenous retroviral envelope protein syncytin 1 and essentially acted as replication-competent retroviruses. The ability to transfer the GFP gene correlated with the levels of syncytin 1 expression in the transduced cells and depended on the fusogenic activity of this protein, substantiating the hypothesis that endogenous syncytin 1 mediates fusion stage in the delivery of extracellular vesicle cargo into target cells. Our findings suggest that testing for replication-competent retroviruses, a routine safety test for transduced cell products in clinical studies, should be also carried out for cell lines generated by retroviral vectors in in vitro studies.

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