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GBM Derived Gangliosides Induce T Cell Apoptosis through Activation of the Caspase Cascade Involving Both the Extrinsic and the Intrinsic Pathway

Previously we demonstrated that human glioblastoma cell lines induce apoptosis in peripheral blood T cells through partial involvement of secreted gangliosides. Here we show that GBM-derived gangliosides induce apoptosis through involvement of the TNF receptor and activation of the caspase cascade....

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Autores principales: Mahata, Barun, Biswas, Soumika, Rayman, Patricia, Chahlavi, Ali, Ko, Jennifer, Bhattacharjee, Ashish, Li, Yu-Teh, Li, Yuntao, Das, Tanya, Sa, Gaurisankar, Raychaudhuri, Baisakhi, Vogelbaum, Michael A., Tannenbaum, Charles, Finke, James H., Biswas, Kaushik
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4520498/
https://www.ncbi.nlm.nih.gov/pubmed/26226135
http://dx.doi.org/10.1371/journal.pone.0134425
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author Mahata, Barun
Biswas, Soumika
Rayman, Patricia
Chahlavi, Ali
Ko, Jennifer
Bhattacharjee, Ashish
Li, Yu-Teh
Li, Yuntao
Das, Tanya
Sa, Gaurisankar
Raychaudhuri, Baisakhi
Vogelbaum, Michael A.
Tannenbaum, Charles
Finke, James H.
Biswas, Kaushik
author_facet Mahata, Barun
Biswas, Soumika
Rayman, Patricia
Chahlavi, Ali
Ko, Jennifer
Bhattacharjee, Ashish
Li, Yu-Teh
Li, Yuntao
Das, Tanya
Sa, Gaurisankar
Raychaudhuri, Baisakhi
Vogelbaum, Michael A.
Tannenbaum, Charles
Finke, James H.
Biswas, Kaushik
author_sort Mahata, Barun
collection PubMed
description Previously we demonstrated that human glioblastoma cell lines induce apoptosis in peripheral blood T cells through partial involvement of secreted gangliosides. Here we show that GBM-derived gangliosides induce apoptosis through involvement of the TNF receptor and activation of the caspase cascade. Culturing T lymphocytes with GBM cell line derived gangliosides (10-20μg/ml) demonstrated increased ROS production as early as 18 hrs as indicated by increased uptake of the dye H(2)DCFDA while western blotting demonstrated mitochondrial damage as evident by cleavage of Bid to t-Bid and by the release of cytochrome-c into the cytosol. Within 48-72 hrs apoptosis was evident by nuclear blebbing, trypan blue positivity and annexinV/7AAD staining. GBM-ganglioside induced activation of the effector caspase-3 along with both initiator caspases (-9 and -8) in T cells while both the caspase-8 and -9 inhibitors were equally effective in blocking apoptosis (60% protection) confirming the role of caspases in the apoptotic process. Ganglioside-induced T cell apoptosis did not involve production of TNF-α since anti-human TNFα antibody was unable to protect T cells from nuclear blebbing and subsequent cell death. However, confocal microscopy demonstrated co-localization of GM2 ganglioside with the TNF receptor and co-immunoprecipitation experiments showed recruitment of death domains FADD and TRADD with the TNF receptor post ganglioside treatment, suggesting direct interaction of gangliosides with the TNF receptor. Further confirmation of the interaction between GM2 and TNFR1 was obtained from confocal microscopy data with wild type and TNFR1 KO (TALEN mediated) Jurkat cells, which clearly demonstrated co-localization of GM2 and TNFR1 in the wild type cells but not in the TNFR1 KO clones. Thus, GBM-ganglioside can mediate T cell apoptosis by interacting with the TNF receptor followed by activation of both the extrinsic and the intrinsic pathway of caspases.
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spelling pubmed-45204982015-08-06 GBM Derived Gangliosides Induce T Cell Apoptosis through Activation of the Caspase Cascade Involving Both the Extrinsic and the Intrinsic Pathway Mahata, Barun Biswas, Soumika Rayman, Patricia Chahlavi, Ali Ko, Jennifer Bhattacharjee, Ashish Li, Yu-Teh Li, Yuntao Das, Tanya Sa, Gaurisankar Raychaudhuri, Baisakhi Vogelbaum, Michael A. Tannenbaum, Charles Finke, James H. Biswas, Kaushik PLoS One Research Article Previously we demonstrated that human glioblastoma cell lines induce apoptosis in peripheral blood T cells through partial involvement of secreted gangliosides. Here we show that GBM-derived gangliosides induce apoptosis through involvement of the TNF receptor and activation of the caspase cascade. Culturing T lymphocytes with GBM cell line derived gangliosides (10-20μg/ml) demonstrated increased ROS production as early as 18 hrs as indicated by increased uptake of the dye H(2)DCFDA while western blotting demonstrated mitochondrial damage as evident by cleavage of Bid to t-Bid and by the release of cytochrome-c into the cytosol. Within 48-72 hrs apoptosis was evident by nuclear blebbing, trypan blue positivity and annexinV/7AAD staining. GBM-ganglioside induced activation of the effector caspase-3 along with both initiator caspases (-9 and -8) in T cells while both the caspase-8 and -9 inhibitors were equally effective in blocking apoptosis (60% protection) confirming the role of caspases in the apoptotic process. Ganglioside-induced T cell apoptosis did not involve production of TNF-α since anti-human TNFα antibody was unable to protect T cells from nuclear blebbing and subsequent cell death. However, confocal microscopy demonstrated co-localization of GM2 ganglioside with the TNF receptor and co-immunoprecipitation experiments showed recruitment of death domains FADD and TRADD with the TNF receptor post ganglioside treatment, suggesting direct interaction of gangliosides with the TNF receptor. Further confirmation of the interaction between GM2 and TNFR1 was obtained from confocal microscopy data with wild type and TNFR1 KO (TALEN mediated) Jurkat cells, which clearly demonstrated co-localization of GM2 and TNFR1 in the wild type cells but not in the TNFR1 KO clones. Thus, GBM-ganglioside can mediate T cell apoptosis by interacting with the TNF receptor followed by activation of both the extrinsic and the intrinsic pathway of caspases. Public Library of Science 2015-07-30 /pmc/articles/PMC4520498/ /pubmed/26226135 http://dx.doi.org/10.1371/journal.pone.0134425 Text en © 2015 Mahata et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Mahata, Barun
Biswas, Soumika
Rayman, Patricia
Chahlavi, Ali
Ko, Jennifer
Bhattacharjee, Ashish
Li, Yu-Teh
Li, Yuntao
Das, Tanya
Sa, Gaurisankar
Raychaudhuri, Baisakhi
Vogelbaum, Michael A.
Tannenbaum, Charles
Finke, James H.
Biswas, Kaushik
GBM Derived Gangliosides Induce T Cell Apoptosis through Activation of the Caspase Cascade Involving Both the Extrinsic and the Intrinsic Pathway
title GBM Derived Gangliosides Induce T Cell Apoptosis through Activation of the Caspase Cascade Involving Both the Extrinsic and the Intrinsic Pathway
title_full GBM Derived Gangliosides Induce T Cell Apoptosis through Activation of the Caspase Cascade Involving Both the Extrinsic and the Intrinsic Pathway
title_fullStr GBM Derived Gangliosides Induce T Cell Apoptosis through Activation of the Caspase Cascade Involving Both the Extrinsic and the Intrinsic Pathway
title_full_unstemmed GBM Derived Gangliosides Induce T Cell Apoptosis through Activation of the Caspase Cascade Involving Both the Extrinsic and the Intrinsic Pathway
title_short GBM Derived Gangliosides Induce T Cell Apoptosis through Activation of the Caspase Cascade Involving Both the Extrinsic and the Intrinsic Pathway
title_sort gbm derived gangliosides induce t cell apoptosis through activation of the caspase cascade involving both the extrinsic and the intrinsic pathway
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4520498/
https://www.ncbi.nlm.nih.gov/pubmed/26226135
http://dx.doi.org/10.1371/journal.pone.0134425
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