Multi-Dimensional Flow Cytometry Analyses Reveal a Dichotomous Role for Nitric Oxide in Melanoma Patients Receiving Immunotherapy
Phenotyping of immune cell subsets in clinical trials is limited to well-defined phenotypes, due to technological limitations of reporting flow cytometry multi-dimensional phenotyping data. We developed a multi-dimensional phenotyping analysis tool and applied it to detect nitric oxide (NO) levels i...
Main Authors: | , , , , , , , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Frontiers Media S.A.
2020-02-01
|
Series: | Frontiers in Immunology |
Subjects: | |
Online Access: | https://www.frontiersin.org/article/10.3389/fimmu.2020.00164/full |
id |
doaj-27868eabfece4beba9ed09c42344dbf3 |
---|---|
record_format |
Article |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Saurabh K. Garg Matthew J. Ott A. G. M. Mostofa Zhihua Chen Y. Ann Chen Jodi Kroeger Biwei Cao Adam W. Mailloux Alisha Agrawal Braydon J. Schaible Amod Sarnaik Amod Sarnaik Jeffrey S. Weber Anders E. Berglund James J. Mulé James J. Mulé Joseph Markowitz Joseph Markowitz Joseph Markowitz |
spellingShingle |
Saurabh K. Garg Matthew J. Ott A. G. M. Mostofa Zhihua Chen Y. Ann Chen Jodi Kroeger Biwei Cao Adam W. Mailloux Alisha Agrawal Braydon J. Schaible Amod Sarnaik Amod Sarnaik Jeffrey S. Weber Anders E. Berglund James J. Mulé James J. Mulé Joseph Markowitz Joseph Markowitz Joseph Markowitz Multi-Dimensional Flow Cytometry Analyses Reveal a Dichotomous Role for Nitric Oxide in Melanoma Patients Receiving Immunotherapy Frontiers in Immunology melanoma nitric oxide immune response ipilimumab flow cytometry |
author_facet |
Saurabh K. Garg Matthew J. Ott A. G. M. Mostofa Zhihua Chen Y. Ann Chen Jodi Kroeger Biwei Cao Adam W. Mailloux Alisha Agrawal Braydon J. Schaible Amod Sarnaik Amod Sarnaik Jeffrey S. Weber Anders E. Berglund James J. Mulé James J. Mulé Joseph Markowitz Joseph Markowitz Joseph Markowitz |
author_sort |
Saurabh K. Garg |
title |
Multi-Dimensional Flow Cytometry Analyses Reveal a Dichotomous Role for Nitric Oxide in Melanoma Patients Receiving Immunotherapy |
title_short |
Multi-Dimensional Flow Cytometry Analyses Reveal a Dichotomous Role for Nitric Oxide in Melanoma Patients Receiving Immunotherapy |
title_full |
Multi-Dimensional Flow Cytometry Analyses Reveal a Dichotomous Role for Nitric Oxide in Melanoma Patients Receiving Immunotherapy |
title_fullStr |
Multi-Dimensional Flow Cytometry Analyses Reveal a Dichotomous Role for Nitric Oxide in Melanoma Patients Receiving Immunotherapy |
title_full_unstemmed |
Multi-Dimensional Flow Cytometry Analyses Reveal a Dichotomous Role for Nitric Oxide in Melanoma Patients Receiving Immunotherapy |
title_sort |
multi-dimensional flow cytometry analyses reveal a dichotomous role for nitric oxide in melanoma patients receiving immunotherapy |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Immunology |
issn |
1664-3224 |
publishDate |
2020-02-01 |
description |
Phenotyping of immune cell subsets in clinical trials is limited to well-defined phenotypes, due to technological limitations of reporting flow cytometry multi-dimensional phenotyping data. We developed a multi-dimensional phenotyping analysis tool and applied it to detect nitric oxide (NO) levels in peripheral blood immune cells before and after adjuvant ipilimumab co-administration with a peptide vaccine in melanoma patients. We analyzed inhibitory and stimulatory markers for immune cell phenotypes that were felt to be important in the NO analysis. The pipeline allows visualization of immune cell phenotypes without knowledge of clustering techniques and to categorize cells by association with relapse-free survival (RFS). Using this analysis, we uncovered the potential for a dichotomous role of NO as a pro- and anti-melanoma factor. NO was found in subsets of immune-suppressor cells associated with shorter-term (≤ 1 year) RFS, whereas NO was also present in immune-stimulatory effector cells obtained from patients with significant longer-term (> 1 year) RFS. These studies provide insights into the cell-specific immunomodulatory role of NO. The methods presented herein can be applied to monitor the pro- and anti-tumor effects of a variety of immune-based therapeutics in cancer patients.Clinical Trial Registration Number: NCT00084656 (https://clinicaltrials.gov/ct2/show/NCT00084656). |
topic |
melanoma nitric oxide immune response ipilimumab flow cytometry |
url |
https://www.frontiersin.org/article/10.3389/fimmu.2020.00164/full |
work_keys_str_mv |
AT saurabhkgarg multidimensionalflowcytometryanalysesrevealadichotomousrolefornitricoxideinmelanomapatientsreceivingimmunotherapy AT matthewjott multidimensionalflowcytometryanalysesrevealadichotomousrolefornitricoxideinmelanomapatientsreceivingimmunotherapy AT agmmostofa multidimensionalflowcytometryanalysesrevealadichotomousrolefornitricoxideinmelanomapatientsreceivingimmunotherapy AT zhihuachen multidimensionalflowcytometryanalysesrevealadichotomousrolefornitricoxideinmelanomapatientsreceivingimmunotherapy AT yannchen multidimensionalflowcytometryanalysesrevealadichotomousrolefornitricoxideinmelanomapatientsreceivingimmunotherapy AT jodikroeger multidimensionalflowcytometryanalysesrevealadichotomousrolefornitricoxideinmelanomapatientsreceivingimmunotherapy AT biweicao multidimensionalflowcytometryanalysesrevealadichotomousrolefornitricoxideinmelanomapatientsreceivingimmunotherapy AT adamwmailloux multidimensionalflowcytometryanalysesrevealadichotomousrolefornitricoxideinmelanomapatientsreceivingimmunotherapy AT alishaagrawal multidimensionalflowcytometryanalysesrevealadichotomousrolefornitricoxideinmelanomapatientsreceivingimmunotherapy AT braydonjschaible multidimensionalflowcytometryanalysesrevealadichotomousrolefornitricoxideinmelanomapatientsreceivingimmunotherapy AT amodsarnaik multidimensionalflowcytometryanalysesrevealadichotomousrolefornitricoxideinmelanomapatientsreceivingimmunotherapy AT amodsarnaik multidimensionalflowcytometryanalysesrevealadichotomousrolefornitricoxideinmelanomapatientsreceivingimmunotherapy AT jeffreysweber multidimensionalflowcytometryanalysesrevealadichotomousrolefornitricoxideinmelanomapatientsreceivingimmunotherapy AT anderseberglund multidimensionalflowcytometryanalysesrevealadichotomousrolefornitricoxideinmelanomapatientsreceivingimmunotherapy AT jamesjmule multidimensionalflowcytometryanalysesrevealadichotomousrolefornitricoxideinmelanomapatientsreceivingimmunotherapy AT jamesjmule multidimensionalflowcytometryanalysesrevealadichotomousrolefornitricoxideinmelanomapatientsreceivingimmunotherapy AT josephmarkowitz multidimensionalflowcytometryanalysesrevealadichotomousrolefornitricoxideinmelanomapatientsreceivingimmunotherapy AT josephmarkowitz multidimensionalflowcytometryanalysesrevealadichotomousrolefornitricoxideinmelanomapatientsreceivingimmunotherapy AT josephmarkowitz multidimensionalflowcytometryanalysesrevealadichotomousrolefornitricoxideinmelanomapatientsreceivingimmunotherapy |
_version_ |
1725103007283019776 |
spelling |
doaj-27868eabfece4beba9ed09c42344dbf32020-11-25T01:27:49ZengFrontiers Media S.A.Frontiers in Immunology1664-32242020-02-011110.3389/fimmu.2020.00164496216Multi-Dimensional Flow Cytometry Analyses Reveal a Dichotomous Role for Nitric Oxide in Melanoma Patients Receiving ImmunotherapySaurabh K. Garg0Matthew J. Ott1A. G. M. Mostofa2Zhihua Chen3Y. Ann Chen4Jodi Kroeger5Biwei Cao6Adam W. Mailloux7Alisha Agrawal8Braydon J. Schaible9Amod Sarnaik10Amod Sarnaik11Jeffrey S. Weber12Anders E. Berglund13James J. Mulé14James J. Mulé15Joseph Markowitz16Joseph Markowitz17Joseph Markowitz18Department of Cutaneous Oncology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, United StatesDepartment of Cutaneous Oncology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, United StatesDepartment of Cutaneous Oncology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, United StatesCancer Informatics Core, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, United StatesDepartment of Biostatistics and Bioinformatics, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, United StatesFlow Cytometry Core, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, United StatesDepartment of Biostatistics and Bioinformatics, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, United StatesDepartment of Immunology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, United StatesDepartment of Oncologic Sciences, USF Health Morsani College of Medicine, University of South Florida, Tampa, FL, United StatesDepartment of Biostatistics and Bioinformatics, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, United StatesDepartment of Cutaneous Oncology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, United StatesDepartment of Oncologic Sciences, USF Health Morsani College of Medicine, University of South Florida, Tampa, FL, United StatesDepartment of Medicine, Laura and Isaac Perlmutter Cancer Center, NYU Langone Health, New York, NY, United StatesDepartment of Biostatistics and Bioinformatics, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, United StatesDepartment of Cutaneous Oncology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, United StatesDepartment of Immunology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, United StatesDepartment of Cutaneous Oncology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, United StatesDepartment of Immunology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, United StatesDepartment of Oncologic Sciences, USF Health Morsani College of Medicine, University of South Florida, Tampa, FL, United StatesPhenotyping of immune cell subsets in clinical trials is limited to well-defined phenotypes, due to technological limitations of reporting flow cytometry multi-dimensional phenotyping data. We developed a multi-dimensional phenotyping analysis tool and applied it to detect nitric oxide (NO) levels in peripheral blood immune cells before and after adjuvant ipilimumab co-administration with a peptide vaccine in melanoma patients. We analyzed inhibitory and stimulatory markers for immune cell phenotypes that were felt to be important in the NO analysis. The pipeline allows visualization of immune cell phenotypes without knowledge of clustering techniques and to categorize cells by association with relapse-free survival (RFS). Using this analysis, we uncovered the potential for a dichotomous role of NO as a pro- and anti-melanoma factor. NO was found in subsets of immune-suppressor cells associated with shorter-term (≤ 1 year) RFS, whereas NO was also present in immune-stimulatory effector cells obtained from patients with significant longer-term (> 1 year) RFS. These studies provide insights into the cell-specific immunomodulatory role of NO. The methods presented herein can be applied to monitor the pro- and anti-tumor effects of a variety of immune-based therapeutics in cancer patients.Clinical Trial Registration Number: NCT00084656 (https://clinicaltrials.gov/ct2/show/NCT00084656).https://www.frontiersin.org/article/10.3389/fimmu.2020.00164/fullmelanomanitric oxideimmune responseipilimumabflow cytometry |