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...

Full description

Bibliographic Details
Main Authors: Menay, Florencia, Herschlik, Leticia, De Toro, Julieta, Cocozza, Federico, Tsacalian, Rodrigo Farid, Gravisaco, Marí­a José, Di Sciullo , Maria Paula, Vendrell, Alejandrina, Waldner, Claudia Inés, Mongini, Claudia
Format: Artículo
Language:Inglés
Published: 2017
Subjects:
Online Access:http://hdl.handle.net/20.500.12123/497
http://journal.frontiersin.org/article/10.3389/fimmu.2017.00286/full
https://doi.org/10.3389/fimmu.2017.00286
_version_ 1855482735395602432
author Menay, Florencia
Herschlik, Leticia
De Toro, Julieta
Cocozza, Federico
Tsacalian, Rodrigo Farid
Gravisaco, Marí­a José
Di Sciullo , Maria Paula
Vendrell, Alejandrina
Waldner, Claudia Inés
Mongini, Claudia
author_browse Cocozza, Federico
De Toro, Julieta
Di Sciullo , Maria Paula
Gravisaco, Marí­a José
Herschlik, Leticia
Menay, Florencia
Mongini, Claudia
Tsacalian, Rodrigo Farid
Vendrell, Alejandrina
Waldner, Claudia Inés
author_facet Menay, Florencia
Herschlik, Leticia
De Toro, Julieta
Cocozza, Federico
Tsacalian, Rodrigo Farid
Gravisaco, Marí­a José
Di Sciullo , Maria Paula
Vendrell, Alejandrina
Waldner, Claudia Inés
Mongini, Claudia
author_sort Menay, Florencia
collection INTA Digital
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 Artículo
id INTA497
institution Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina)
language Inglés
publishDate 2017
publishDateRange 2017
publishDateSort 2017
record_format dspace
spelling INTA4972017-12-26T12:07:04Z 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 Farid Gravisaco, Marí­a José Di Sciullo , Maria Paula Vendrell, Alejandrina Waldner, Claudia Inés Mongini, Claudia Respuesta inmunológica Linfoma Ascitis Ratón Linfocitos-t Neoplasmas Immune Response Lymphoma Mice T-lymphocytes 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. Inst. de Biotecnología Fil: Menay, Florencia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Estudios Farmacológicos y Botánicos; Argentina. Universidad de Buenos. Facultad de Medicina, Aires; Argentina Fil: Herschlik, Leticia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Estudios Farmacológicos y Botánicos; Argentina. Universidad de Buenos. Facultad de Medicina, Aires; Argentina Fil: Cocozza, Federico. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Estudios Farmacológicos y Botánicos; Argentina. Universidad de Buenos. Facultad de Medicina, Aires; Argentina Fil: Tsacalian, Rodrigo Farid. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Estudios Farmacológicos y Botánicos; Argentina. Universidad de Buenos. Facultad de Medicina, Aires; Argentina Fil: Gravisaco, Marí­a José. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Di Sciullo, Maria Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Estudios Farmacológicos y Botánicos; Argentina. Universidad de Buenos. Facultad de Medicina; Argentina Fil: Vendrell, Alejandrina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Estudios Farmacológicos y Botánicos; Argentina. Universidad de Buenos. Facultad de Medicina, Aires; Argentina Fil: Waldner, Claudia Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Estudios Farmacológicos y Botánicos; Argentina. Universidad de Buenos. Facultad de Medicina, Aires; Argentina Fil: Mongini, Claudia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Estudios Farmacológicos y Botánicos; Argentina. Universidad de Buenos. Facultad de Medicina; Argentina 2017-06-28T14:46:22Z 2017-06-28T14:46:22Z 2017-03-16 info:ar-repo/semantics/artículo info:eu-repo/semantics/publishedVersion info:eu-repo/semantics/article http://hdl.handle.net/20.500.12123/497 http://journal.frontiersin.org/article/10.3389/fimmu.2017.00286/full https://doi.org/10.3389/fimmu.2017.00286 eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf Frontiers in immunology 8 : 1-14. (March 2017)
spellingShingle Respuesta inmunológica
Linfoma
Ascitis
Ratón
Linfocitos-t
Neoplasmas
Immune Response
Lymphoma
Mice
T-lymphocytes
Menay, Florencia
Herschlik, Leticia
De Toro, Julieta
Cocozza, Federico
Tsacalian, Rodrigo Farid
Gravisaco, Marí­a José
Di Sciullo , Maria Paula
Vendrell, Alejandrina
Waldner, Claudia Inés
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 Respuesta inmunológica
Linfoma
Ascitis
Ratón
Linfocitos-t
Neoplasmas
Immune Response
Lymphoma
Mice
T-lymphocytes
url http://hdl.handle.net/20.500.12123/497
http://journal.frontiersin.org/article/10.3389/fimmu.2017.00286/full
https://doi.org/10.3389/fimmu.2017.00286
work_keys_str_mv AT menayflorencia exosomesisolatedfromascitesoftcelllymphomabearingmiceexpressingsurfacecd24andhsp90induceatumorspecificimmuneresponse
AT herschlikleticia exosomesisolatedfromascitesoftcelllymphomabearingmiceexpressingsurfacecd24andhsp90induceatumorspecificimmuneresponse
AT detorojulieta exosomesisolatedfromascitesoftcelllymphomabearingmiceexpressingsurfacecd24andhsp90induceatumorspecificimmuneresponse
AT cocozzafederico exosomesisolatedfromascitesoftcelllymphomabearingmiceexpressingsurfacecd24andhsp90induceatumorspecificimmuneresponse
AT tsacalianrodrigofarid exosomesisolatedfromascitesoftcelllymphomabearingmiceexpressingsurfacecd24andhsp90induceatumorspecificimmuneresponse
AT gravisacomariajose exosomesisolatedfromascitesoftcelllymphomabearingmiceexpressingsurfacecd24andhsp90induceatumorspecificimmuneresponse
AT disciullomariapaula exosomesisolatedfromascitesoftcelllymphomabearingmiceexpressingsurfacecd24andhsp90induceatumorspecificimmuneresponse
AT vendrellalejandrina exosomesisolatedfromascitesoftcelllymphomabearingmiceexpressingsurfacecd24andhsp90induceatumorspecificimmuneresponse
AT waldnerclaudiaines exosomesisolatedfromascitesoftcelllymphomabearingmiceexpressingsurfacecd24andhsp90induceatumorspecificimmuneresponse
AT monginiclaudia exosomesisolatedfromascitesoftcelllymphomabearingmiceexpressingsurfacecd24andhsp90induceatumorspecificimmuneresponse