Cargando…

Exosomes Isolated from Ascites of T-Cell Lymphoma-Bearing Mice Expressing Surface CD24 and HSP-90 Induce a Tumor-Specific Immune Response

Extracellular vesicles (EVs), including endosome-derived nanovesicles (exosomes), are involved in cell–cell communication. Through transfer of their molecular contents, extracellular nanovesicles can alter the function of recipient cells. Due to these characteristics, EVs have shown potential as a n...

Descripción completa

Detalles Bibliográficos
Autores principales: Menay, Florencia, Herschlik, Leticia, De Toro, Julieta, Cocozza, Federico, Tsacalian, Rodrigo, Gravisaco, María José, Di Sciullo, María Paula, Vendrell, Alejandrina, Waldner, Claudia I., Mongini, Claudia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5352668/
https://www.ncbi.nlm.nih.gov/pubmed/28360912
http://dx.doi.org/10.3389/fimmu.2017.00286
_version_ 1782514989339246592
author Menay, Florencia
Herschlik, Leticia
De Toro, Julieta
Cocozza, Federico
Tsacalian, Rodrigo
Gravisaco, María José
Di Sciullo, María Paula
Vendrell, Alejandrina
Waldner, Claudia I.
Mongini, Claudia
author_facet Menay, Florencia
Herschlik, Leticia
De Toro, Julieta
Cocozza, Federico
Tsacalian, Rodrigo
Gravisaco, María José
Di Sciullo, María Paula
Vendrell, Alejandrina
Waldner, Claudia I.
Mongini, Claudia
author_sort Menay, Florencia
collection PubMed
description Extracellular vesicles (EVs), including endosome-derived nanovesicles (exosomes), are involved in cell–cell communication. Through transfer of their molecular contents, extracellular nanovesicles can alter the function of recipient cells. Due to these characteristics, EVs have shown potential as a new alternative for cancer immunotherapy. Tumor exosomes isolated from malignant ascites can activate dendritic cells, thereby priming the immune system to recognize and kill cancer cells. However, a suppressive role on tumor immune response has also been reported, suggesting that the neoplastic stage of carcinogenesis and the microenvironment where tumor cells grow may influence the amount of EVs released by the cell. This neoplastic stage and microenvironment may also impact EVs’ components such as proteins and miRNA, determining their biological behavior. Most T-cell lymphomas have an aggressive clinical course and poor prognosis. Consequently, complementary alternative therapies are needed to improve the survival rates achieved with conventional treatments. In this work, we have characterized EVs isolated from ascites of mice bearing a very aggressive murine T-cell lymphoma and have studied their immunogenic properties. Small EVs were isolated by differential centrifugation, ultrafiltration, and ultracentrifugation at 100,000 × g on a sucrose cushion. The EVs were defined as exosomes by their morphology and size analyzed by electron microscopy, their floating density on a sucrose gradient, as well as their expression of endosome marker proteins ALIX, TSG-101; the tetraspanins CD63, CD9, and CD81. In addition, they contain tumor antigens, the marker for malignancy CD24, the heat shock protein HSP-70, and an unusual surface expression of HSP-90 was demonstrated. The administration of EVs isolated from ascites (EVs A) into naïve-syngeneic mice induced both humoral and cellular immune responses that allowed the rejection of subsequent tumor challenges. However, the immunization had no effect on a non-related mammary adenocarcinoma, demonstrating that the immune response elicited was specific and also it induced immune memory. In vitro analysis demonstrated that T-cells from EVs A-immunized mice secrete IFN-γ in response to tumor stimulation. Furthermore, tumor-specific CD4+ and CD8+ IFN-γ secreting cells could be efficiently expanded from mice immunized with EVs A, showing that a T helper 1 response is involved in tumor rejection. Our findings confirm exosomes as promising defined acellular tumor antigens for the development of an antitumor vaccine.
format Online
Article
Text
id pubmed-5352668
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-53526682017-03-30 Exosomes Isolated from Ascites of T-Cell Lymphoma-Bearing Mice Expressing Surface CD24 and HSP-90 Induce a Tumor-Specific Immune Response Menay, Florencia Herschlik, Leticia De Toro, Julieta Cocozza, Federico Tsacalian, Rodrigo Gravisaco, María José Di Sciullo, María Paula Vendrell, Alejandrina Waldner, Claudia I. Mongini, Claudia Front Immunol Immunology Extracellular vesicles (EVs), including endosome-derived nanovesicles (exosomes), are involved in cell–cell communication. Through transfer of their molecular contents, extracellular nanovesicles can alter the function of recipient cells. Due to these characteristics, EVs have shown potential as a new alternative for cancer immunotherapy. Tumor exosomes isolated from malignant ascites can activate dendritic cells, thereby priming the immune system to recognize and kill cancer cells. However, a suppressive role on tumor immune response has also been reported, suggesting that the neoplastic stage of carcinogenesis and the microenvironment where tumor cells grow may influence the amount of EVs released by the cell. This neoplastic stage and microenvironment may also impact EVs’ components such as proteins and miRNA, determining their biological behavior. Most T-cell lymphomas have an aggressive clinical course and poor prognosis. Consequently, complementary alternative therapies are needed to improve the survival rates achieved with conventional treatments. In this work, we have characterized EVs isolated from ascites of mice bearing a very aggressive murine T-cell lymphoma and have studied their immunogenic properties. Small EVs were isolated by differential centrifugation, ultrafiltration, and ultracentrifugation at 100,000 × g on a sucrose cushion. The EVs were defined as exosomes by their morphology and size analyzed by electron microscopy, their floating density on a sucrose gradient, as well as their expression of endosome marker proteins ALIX, TSG-101; the tetraspanins CD63, CD9, and CD81. In addition, they contain tumor antigens, the marker for malignancy CD24, the heat shock protein HSP-70, and an unusual surface expression of HSP-90 was demonstrated. The administration of EVs isolated from ascites (EVs A) into naïve-syngeneic mice induced both humoral and cellular immune responses that allowed the rejection of subsequent tumor challenges. However, the immunization had no effect on a non-related mammary adenocarcinoma, demonstrating that the immune response elicited was specific and also it induced immune memory. In vitro analysis demonstrated that T-cells from EVs A-immunized mice secrete IFN-γ in response to tumor stimulation. Furthermore, tumor-specific CD4+ and CD8+ IFN-γ secreting cells could be efficiently expanded from mice immunized with EVs A, showing that a T helper 1 response is involved in tumor rejection. Our findings confirm exosomes as promising defined acellular tumor antigens for the development of an antitumor vaccine. Frontiers Media S.A. 2017-03-16 /pmc/articles/PMC5352668/ /pubmed/28360912 http://dx.doi.org/10.3389/fimmu.2017.00286 Text en Copyright © 2017 Menay, Herschlik, De Toro, Cocozza, Tsacalian, Gravisaco, Di Sciullo, Vendrell, Waldner and Mongini. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Immunology
Menay, Florencia
Herschlik, Leticia
De Toro, Julieta
Cocozza, Federico
Tsacalian, Rodrigo
Gravisaco, María José
Di Sciullo, María Paula
Vendrell, Alejandrina
Waldner, Claudia I.
Mongini, Claudia
Exosomes Isolated from Ascites of T-Cell Lymphoma-Bearing Mice Expressing Surface CD24 and HSP-90 Induce a Tumor-Specific Immune Response
title Exosomes Isolated from Ascites of T-Cell Lymphoma-Bearing Mice Expressing Surface CD24 and HSP-90 Induce a Tumor-Specific Immune Response
title_full Exosomes Isolated from Ascites of T-Cell Lymphoma-Bearing Mice Expressing Surface CD24 and HSP-90 Induce a Tumor-Specific Immune Response
title_fullStr Exosomes Isolated from Ascites of T-Cell Lymphoma-Bearing Mice Expressing Surface CD24 and HSP-90 Induce a Tumor-Specific Immune Response
title_full_unstemmed Exosomes Isolated from Ascites of T-Cell Lymphoma-Bearing Mice Expressing Surface CD24 and HSP-90 Induce a Tumor-Specific Immune Response
title_short Exosomes Isolated from Ascites of T-Cell Lymphoma-Bearing Mice Expressing Surface CD24 and HSP-90 Induce a Tumor-Specific Immune Response
title_sort exosomes isolated from ascites of t-cell lymphoma-bearing mice expressing surface cd24 and hsp-90 induce a tumor-specific immune response
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5352668/
https://www.ncbi.nlm.nih.gov/pubmed/28360912
http://dx.doi.org/10.3389/fimmu.2017.00286
work_keys_str_mv AT menayflorencia exosomesisolatedfromascitesoftcelllymphomabearingmiceexpressingsurfacecd24andhsp90induceatumorspecificimmuneresponse
AT herschlikleticia exosomesisolatedfromascitesoftcelllymphomabearingmiceexpressingsurfacecd24andhsp90induceatumorspecificimmuneresponse
AT detorojulieta exosomesisolatedfromascitesoftcelllymphomabearingmiceexpressingsurfacecd24andhsp90induceatumorspecificimmuneresponse
AT cocozzafederico exosomesisolatedfromascitesoftcelllymphomabearingmiceexpressingsurfacecd24andhsp90induceatumorspecificimmuneresponse
AT tsacalianrodrigo exosomesisolatedfromascitesoftcelllymphomabearingmiceexpressingsurfacecd24andhsp90induceatumorspecificimmuneresponse
AT gravisacomariajose exosomesisolatedfromascitesoftcelllymphomabearingmiceexpressingsurfacecd24andhsp90induceatumorspecificimmuneresponse
AT disciullomariapaula exosomesisolatedfromascitesoftcelllymphomabearingmiceexpressingsurfacecd24andhsp90induceatumorspecificimmuneresponse
AT vendrellalejandrina exosomesisolatedfromascitesoftcelllymphomabearingmiceexpressingsurfacecd24andhsp90induceatumorspecificimmuneresponse
AT waldnerclaudiai exosomesisolatedfromascitesoftcelllymphomabearingmiceexpressingsurfacecd24andhsp90induceatumorspecificimmuneresponse
AT monginiclaudia exosomesisolatedfromascitesoftcelllymphomabearingmiceexpressingsurfacecd24andhsp90induceatumorspecificimmuneresponse