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The engagement of CTLA-4 on primary melanoma cell lines induces antibody-dependent cellular cytotoxicity and TNF-α production
BACKGROUND: CTLA-4 (Cytotoxic T lymphocyte antigen-4) is traditionally known as a negative regulator of T cell activation. The blocking of CTLA-4 using human monoclonal antibodies, such as Ipilimumab, is currently used to relieve CTLA-4-mediated inhibition of anti-tumor immune response in metastatic...
Autores principales: | , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3663700/ https://www.ncbi.nlm.nih.gov/pubmed/23634660 http://dx.doi.org/10.1186/1479-5876-11-108 |
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author | Laurent, Stefania Queirolo, Paola Boero, Silvia Salvi, Sandra Piccioli, Patrizia Boccardo, Simona Minghelli, Simona Morabito, Anna Fontana, Vincenzo Pietra, Gabriella Carrega, Paolo Ferrari, Nicoletta Tosetti, Francesca Chang, Lung-Ji Mingari, Maria Cristina Ferlazzo, Guido Poggi, Alessandro Pistillo, Maria Pia |
author_facet | Laurent, Stefania Queirolo, Paola Boero, Silvia Salvi, Sandra Piccioli, Patrizia Boccardo, Simona Minghelli, Simona Morabito, Anna Fontana, Vincenzo Pietra, Gabriella Carrega, Paolo Ferrari, Nicoletta Tosetti, Francesca Chang, Lung-Ji Mingari, Maria Cristina Ferlazzo, Guido Poggi, Alessandro Pistillo, Maria Pia |
author_sort | Laurent, Stefania |
collection | PubMed |
description | BACKGROUND: CTLA-4 (Cytotoxic T lymphocyte antigen-4) is traditionally known as a negative regulator of T cell activation. The blocking of CTLA-4 using human monoclonal antibodies, such as Ipilimumab, is currently used to relieve CTLA-4-mediated inhibition of anti-tumor immune response in metastatic melanoma. Herein, we have analyzed CTLA-4 expression and Ipilimumab reactivity on melanoma cell lines and tumor tissues from cutaneous melanoma patients. Then, we investigated whether Ipilimumab can trigger innate immunity in terms of antibody dependent cellular cytotoxicity (ADCC) or Tumor Necrosis Factor (TNF)-α release. Finally, a xenograft murine model was set up to determine in vivo the effects of Ipilimumab and NK cells on melanoma. METHODS: CTLA-4 expression and Ipilimumab reactivity were analyzed on 17 melanoma cell lines (14 primary and 3 long-term cell lines) by cytofluorimetry and on 33 melanoma tissues by immunohistochemistry. CTLA-4 transcripts were analyzed by quantitative RT-PCR. Soluble CTLA-4 and TNF-α were tested by ELISA. Peripheral blood mononuclear cells (PBMC), NK and γδT cells were tested in ADCC assay with Ipilimumab and melanoma cell lines. TNF-α release was analyzed in NK-melanoma cell co-cultures in the presence of ipilimumab. In vivo experiments of xenotransplantation were carried out in NOD/SCID mice. Results were analyzed using unpaired Student’s t-test. RESULTS: All melanoma cell lines expressed mRNA and cytoplasmic CTLA-4 but surface reactivity with Ipilimumab was quite heterogeneous. Accordingly, about 2/3 of melanoma specimens expressed CTLA-4 at different level of intensity. Ipilimumab triggered, via FcγReceptorIIIA (CD16), ex vivo NK cells as well as PBMC, IL-2 activated NK and γδT cells to ADCC of CTLA-4(+) melanoma cells. No ADCC was detected upon interaction with CTLA-4(-) FO-1 melanoma cell line. TNF-α was released upon interaction of NK cells with CTLA-4(+) melanoma cell lines. Remarkably, Ipilimumab neither affected proliferation and viability nor triggered ADCC of CTLA-4(+) T lymphocytes. In a chimeric murine xenograft model, the co-engraftment of Ipilimumab-treated melanoma cells with human allogeneic NK cells delayed and significantly reduced tumor growth, as compared to mice receiving control xenografts. CONCLUSIONS: Our studies demonstrate that Ipilimumab triggers effector lymphocytes to cytotoxicity and TNF-α release. These findings suggest that Ipilimumab, besides blocking CTLA-4, can directly activate the elimination of CTLA-4(+) melanomas. |
format | Online Article Text |
id | pubmed-3663700 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-36637002013-05-25 The engagement of CTLA-4 on primary melanoma cell lines induces antibody-dependent cellular cytotoxicity and TNF-α production Laurent, Stefania Queirolo, Paola Boero, Silvia Salvi, Sandra Piccioli, Patrizia Boccardo, Simona Minghelli, Simona Morabito, Anna Fontana, Vincenzo Pietra, Gabriella Carrega, Paolo Ferrari, Nicoletta Tosetti, Francesca Chang, Lung-Ji Mingari, Maria Cristina Ferlazzo, Guido Poggi, Alessandro Pistillo, Maria Pia J Transl Med Research BACKGROUND: CTLA-4 (Cytotoxic T lymphocyte antigen-4) is traditionally known as a negative regulator of T cell activation. The blocking of CTLA-4 using human monoclonal antibodies, such as Ipilimumab, is currently used to relieve CTLA-4-mediated inhibition of anti-tumor immune response in metastatic melanoma. Herein, we have analyzed CTLA-4 expression and Ipilimumab reactivity on melanoma cell lines and tumor tissues from cutaneous melanoma patients. Then, we investigated whether Ipilimumab can trigger innate immunity in terms of antibody dependent cellular cytotoxicity (ADCC) or Tumor Necrosis Factor (TNF)-α release. Finally, a xenograft murine model was set up to determine in vivo the effects of Ipilimumab and NK cells on melanoma. METHODS: CTLA-4 expression and Ipilimumab reactivity were analyzed on 17 melanoma cell lines (14 primary and 3 long-term cell lines) by cytofluorimetry and on 33 melanoma tissues by immunohistochemistry. CTLA-4 transcripts were analyzed by quantitative RT-PCR. Soluble CTLA-4 and TNF-α were tested by ELISA. Peripheral blood mononuclear cells (PBMC), NK and γδT cells were tested in ADCC assay with Ipilimumab and melanoma cell lines. TNF-α release was analyzed in NK-melanoma cell co-cultures in the presence of ipilimumab. In vivo experiments of xenotransplantation were carried out in NOD/SCID mice. Results were analyzed using unpaired Student’s t-test. RESULTS: All melanoma cell lines expressed mRNA and cytoplasmic CTLA-4 but surface reactivity with Ipilimumab was quite heterogeneous. Accordingly, about 2/3 of melanoma specimens expressed CTLA-4 at different level of intensity. Ipilimumab triggered, via FcγReceptorIIIA (CD16), ex vivo NK cells as well as PBMC, IL-2 activated NK and γδT cells to ADCC of CTLA-4(+) melanoma cells. No ADCC was detected upon interaction with CTLA-4(-) FO-1 melanoma cell line. TNF-α was released upon interaction of NK cells with CTLA-4(+) melanoma cell lines. Remarkably, Ipilimumab neither affected proliferation and viability nor triggered ADCC of CTLA-4(+) T lymphocytes. In a chimeric murine xenograft model, the co-engraftment of Ipilimumab-treated melanoma cells with human allogeneic NK cells delayed and significantly reduced tumor growth, as compared to mice receiving control xenografts. CONCLUSIONS: Our studies demonstrate that Ipilimumab triggers effector lymphocytes to cytotoxicity and TNF-α release. These findings suggest that Ipilimumab, besides blocking CTLA-4, can directly activate the elimination of CTLA-4(+) melanomas. BioMed Central 2013-05-01 /pmc/articles/PMC3663700/ /pubmed/23634660 http://dx.doi.org/10.1186/1479-5876-11-108 Text en Copyright © 2013 Laurent et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Laurent, Stefania Queirolo, Paola Boero, Silvia Salvi, Sandra Piccioli, Patrizia Boccardo, Simona Minghelli, Simona Morabito, Anna Fontana, Vincenzo Pietra, Gabriella Carrega, Paolo Ferrari, Nicoletta Tosetti, Francesca Chang, Lung-Ji Mingari, Maria Cristina Ferlazzo, Guido Poggi, Alessandro Pistillo, Maria Pia The engagement of CTLA-4 on primary melanoma cell lines induces antibody-dependent cellular cytotoxicity and TNF-α production |
title | The engagement of CTLA-4 on primary melanoma cell lines induces antibody-dependent cellular cytotoxicity and TNF-α production |
title_full | The engagement of CTLA-4 on primary melanoma cell lines induces antibody-dependent cellular cytotoxicity and TNF-α production |
title_fullStr | The engagement of CTLA-4 on primary melanoma cell lines induces antibody-dependent cellular cytotoxicity and TNF-α production |
title_full_unstemmed | The engagement of CTLA-4 on primary melanoma cell lines induces antibody-dependent cellular cytotoxicity and TNF-α production |
title_short | The engagement of CTLA-4 on primary melanoma cell lines induces antibody-dependent cellular cytotoxicity and TNF-α production |
title_sort | engagement of ctla-4 on primary melanoma cell lines induces antibody-dependent cellular cytotoxicity and tnf-α production |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3663700/ https://www.ncbi.nlm.nih.gov/pubmed/23634660 http://dx.doi.org/10.1186/1479-5876-11-108 |
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