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Isolated Limb Perfusion With Melphalan Triggers Immune Activation in Melanoma Patients

Hyperthermic isolated limb perfusion with melphalan (M-ILP) is a treatment option for melanoma patients with metastases confined to the limbs. This study aimed at defining the role of cellular immunity for the clinical response to M-ILP in melanoma patients. It was observed that patients with enhanc...

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Autores principales: Johansson, Junko, Kiffin, Roberta, Andersson, Annica, Lindnér, Per, Naredi, Peter L., Olofsson Bagge, Roger, Martner, Anna
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6286961/
https://www.ncbi.nlm.nih.gov/pubmed/30560089
http://dx.doi.org/10.3389/fonc.2018.00570
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author Johansson, Junko
Kiffin, Roberta
Andersson, Annica
Lindnér, Per
Naredi, Peter L.
Olofsson Bagge, Roger
Martner, Anna
author_facet Johansson, Junko
Kiffin, Roberta
Andersson, Annica
Lindnér, Per
Naredi, Peter L.
Olofsson Bagge, Roger
Martner, Anna
author_sort Johansson, Junko
collection PubMed
description Hyperthermic isolated limb perfusion with melphalan (M-ILP) is a treatment option for melanoma patients with metastases confined to the limbs. This study aimed at defining the role of cellular immunity for the clinical response to M-ILP in melanoma patients. It was observed that patients with enhanced cytotoxic CD8(+) T cell reactivity to common antigens (HCMV/EBV/influenza virus) prior to M-ILP were more likely to achieve a complete disappearance of macroscopic tumors (complete response). Following M-ILP treatment, the proportions of CD16(+) intermediate and non-classical monocytes in peripheral blood were significantly enhanced along with induction of HLA-DR on CD4(+) and CD8(+) T cells. For further studies of the mechanism behind melphalan-induced immune activation an in vitro model, aiming at mimicking the clinical M-ILP protocol, was established, where PBMCs were co-cultured with melanoma cells, which had been pre-exposed to melphalan under mild hyperthermia. Upon exposure to melphalan, melanoma cells showed increased expression of immune-related markers including MHC class I and Hsp70. Moreover, when the melphalan-treated melanoma cells were co-cultured with PBMCs, this triggered an increased proportion of CD33(+)CD14(+)CD16(++) non-classical monocytes among the PBMCs. Furthermore, the melphalan-treated melanoma cells stimulated the expansion of CD8(+) T cells in the co-cultured PBMCs. These cells produced enhanced levels of IFN-γ and granzyme B and were capable of killing melanoma cells. To further verify an immunogenic role of melphalan, mice were vaccinated with melphalan-exposed murine melanoma cells. When challenged with live melanoma cells, vaccinated mice showed reduced tumor growth and enhanced infiltration of tumor-specific T cells into tumors. We conclude that melphalan-exposed melanoma cells trigger expansion of CD16(+) monocytes and activate cytotoxic T cells and that these events may contribute to the antitumoral efficacy of M-ILP.
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spelling pubmed-62869612018-12-17 Isolated Limb Perfusion With Melphalan Triggers Immune Activation in Melanoma Patients Johansson, Junko Kiffin, Roberta Andersson, Annica Lindnér, Per Naredi, Peter L. Olofsson Bagge, Roger Martner, Anna Front Oncol Oncology Hyperthermic isolated limb perfusion with melphalan (M-ILP) is a treatment option for melanoma patients with metastases confined to the limbs. This study aimed at defining the role of cellular immunity for the clinical response to M-ILP in melanoma patients. It was observed that patients with enhanced cytotoxic CD8(+) T cell reactivity to common antigens (HCMV/EBV/influenza virus) prior to M-ILP were more likely to achieve a complete disappearance of macroscopic tumors (complete response). Following M-ILP treatment, the proportions of CD16(+) intermediate and non-classical monocytes in peripheral blood were significantly enhanced along with induction of HLA-DR on CD4(+) and CD8(+) T cells. For further studies of the mechanism behind melphalan-induced immune activation an in vitro model, aiming at mimicking the clinical M-ILP protocol, was established, where PBMCs were co-cultured with melanoma cells, which had been pre-exposed to melphalan under mild hyperthermia. Upon exposure to melphalan, melanoma cells showed increased expression of immune-related markers including MHC class I and Hsp70. Moreover, when the melphalan-treated melanoma cells were co-cultured with PBMCs, this triggered an increased proportion of CD33(+)CD14(+)CD16(++) non-classical monocytes among the PBMCs. Furthermore, the melphalan-treated melanoma cells stimulated the expansion of CD8(+) T cells in the co-cultured PBMCs. These cells produced enhanced levels of IFN-γ and granzyme B and were capable of killing melanoma cells. To further verify an immunogenic role of melphalan, mice were vaccinated with melphalan-exposed murine melanoma cells. When challenged with live melanoma cells, vaccinated mice showed reduced tumor growth and enhanced infiltration of tumor-specific T cells into tumors. We conclude that melphalan-exposed melanoma cells trigger expansion of CD16(+) monocytes and activate cytotoxic T cells and that these events may contribute to the antitumoral efficacy of M-ILP. Frontiers Media S.A. 2018-12-03 /pmc/articles/PMC6286961/ /pubmed/30560089 http://dx.doi.org/10.3389/fonc.2018.00570 Text en Copyright © 2018 Johansson, Kiffin, Andersson, Lindnér, Naredi, Olofsson Bagge and Martner. 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) and the copyright owner(s) 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 Oncology
Johansson, Junko
Kiffin, Roberta
Andersson, Annica
Lindnér, Per
Naredi, Peter L.
Olofsson Bagge, Roger
Martner, Anna
Isolated Limb Perfusion With Melphalan Triggers Immune Activation in Melanoma Patients
title Isolated Limb Perfusion With Melphalan Triggers Immune Activation in Melanoma Patients
title_full Isolated Limb Perfusion With Melphalan Triggers Immune Activation in Melanoma Patients
title_fullStr Isolated Limb Perfusion With Melphalan Triggers Immune Activation in Melanoma Patients
title_full_unstemmed Isolated Limb Perfusion With Melphalan Triggers Immune Activation in Melanoma Patients
title_short Isolated Limb Perfusion With Melphalan Triggers Immune Activation in Melanoma Patients
title_sort isolated limb perfusion with melphalan triggers immune activation in melanoma patients
topic Oncology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6286961/
https://www.ncbi.nlm.nih.gov/pubmed/30560089
http://dx.doi.org/10.3389/fonc.2018.00570
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