Cargando…

Multiplex Immunofluorescence Tyramide Signal Amplification for Immune Cell Profiling of Paraffin-Embedded Tumor Tissues

Every day, more evidence is revealed regarding the importance of the relationship between the response to cancer immunotherapy and the cancer immune microenvironment. It is well established that a profound characterization of the immune microenvironment is needed to identify prognostic and predictiv...

Descripción completa

Detalles Bibliográficos
Autores principales: Hernandez, Sharia, Rojas, Frank, Laberiano, Caddie, Lazcano, Rossana, Wistuba, Ignacio, Parra, Edwin Roger
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8118604/
https://www.ncbi.nlm.nih.gov/pubmed/33996912
http://dx.doi.org/10.3389/fmolb.2021.667067
_version_ 1783691779961257984
author Hernandez, Sharia
Rojas, Frank
Laberiano, Caddie
Lazcano, Rossana
Wistuba, Ignacio
Parra, Edwin Roger
author_facet Hernandez, Sharia
Rojas, Frank
Laberiano, Caddie
Lazcano, Rossana
Wistuba, Ignacio
Parra, Edwin Roger
author_sort Hernandez, Sharia
collection PubMed
description Every day, more evidence is revealed regarding the importance of the relationship between the response to cancer immunotherapy and the cancer immune microenvironment. It is well established that a profound characterization of the immune microenvironment is needed to identify prognostic and predictive immune biomarkers. To this end, we find phenotyping cells by multiplex immunofluorescence (mIF) a powerful and useful tool to identify cell types in biopsy specimens. Here, we describe the use of mIF tyramide signal amplification for labeling up to eight markers on a single slide of formalin-fixed, paraffin-embedded tumor tissue to phenotype immune cells in tumor tissues. Different panels show different markers, and the different panels can be used to characterize immune cells and relevant checkpoint proteins. The panel design depends on the research hypothesis, the cell population of interest, or the treatment under investigation. To phenotype the cells, image analysis software is used to identify individual marker expression or specific co-expression markers, which can differentiate already selected phenotypes. The individual-markers approach identifies a broad number of cell phenotypes, including rare cells, which may be helpful in a tumor microenvironment study. To accurately interpret results, it is important to recognize which receptors are expressed on different cell types and their typical location (i.e., nuclear, membrane, and/or cytoplasm). Furthermore, the amplification system of mIF may allow us to see weak marker signals, such as programmed cell death ligand 1, more easily than they are seen with single-marker immunohistochemistry (IHC) labeling. Finally, mIF technologies are promising resources for discovery of novel cancer immunotherapies and related biomarkers. In contrast with conventional IHC, which permits only the labeling of one single marker per tissue sample, mIF can detect multiple markers from a single tissue sample, and at the same time, deliver extensive information about the cell phenotypes composition and their spatial localization. In this matter, the phenotyping process is critical and must be done accurately by a highly trained personal with knowledge of immune cell protein expression and tumor pathology.
format Online
Article
Text
id pubmed-8118604
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-81186042021-05-14 Multiplex Immunofluorescence Tyramide Signal Amplification for Immune Cell Profiling of Paraffin-Embedded Tumor Tissues Hernandez, Sharia Rojas, Frank Laberiano, Caddie Lazcano, Rossana Wistuba, Ignacio Parra, Edwin Roger Front Mol Biosci Molecular Biosciences Every day, more evidence is revealed regarding the importance of the relationship between the response to cancer immunotherapy and the cancer immune microenvironment. It is well established that a profound characterization of the immune microenvironment is needed to identify prognostic and predictive immune biomarkers. To this end, we find phenotyping cells by multiplex immunofluorescence (mIF) a powerful and useful tool to identify cell types in biopsy specimens. Here, we describe the use of mIF tyramide signal amplification for labeling up to eight markers on a single slide of formalin-fixed, paraffin-embedded tumor tissue to phenotype immune cells in tumor tissues. Different panels show different markers, and the different panels can be used to characterize immune cells and relevant checkpoint proteins. The panel design depends on the research hypothesis, the cell population of interest, or the treatment under investigation. To phenotype the cells, image analysis software is used to identify individual marker expression or specific co-expression markers, which can differentiate already selected phenotypes. The individual-markers approach identifies a broad number of cell phenotypes, including rare cells, which may be helpful in a tumor microenvironment study. To accurately interpret results, it is important to recognize which receptors are expressed on different cell types and their typical location (i.e., nuclear, membrane, and/or cytoplasm). Furthermore, the amplification system of mIF may allow us to see weak marker signals, such as programmed cell death ligand 1, more easily than they are seen with single-marker immunohistochemistry (IHC) labeling. Finally, mIF technologies are promising resources for discovery of novel cancer immunotherapies and related biomarkers. In contrast with conventional IHC, which permits only the labeling of one single marker per tissue sample, mIF can detect multiple markers from a single tissue sample, and at the same time, deliver extensive information about the cell phenotypes composition and their spatial localization. In this matter, the phenotyping process is critical and must be done accurately by a highly trained personal with knowledge of immune cell protein expression and tumor pathology. Frontiers Media S.A. 2021-04-29 /pmc/articles/PMC8118604/ /pubmed/33996912 http://dx.doi.org/10.3389/fmolb.2021.667067 Text en Copyright © 2021 Hernandez, Rojas, Laberiano, Lazcano, Wistuba and Parra. https://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 Molecular Biosciences
Hernandez, Sharia
Rojas, Frank
Laberiano, Caddie
Lazcano, Rossana
Wistuba, Ignacio
Parra, Edwin Roger
Multiplex Immunofluorescence Tyramide Signal Amplification for Immune Cell Profiling of Paraffin-Embedded Tumor Tissues
title Multiplex Immunofluorescence Tyramide Signal Amplification for Immune Cell Profiling of Paraffin-Embedded Tumor Tissues
title_full Multiplex Immunofluorescence Tyramide Signal Amplification for Immune Cell Profiling of Paraffin-Embedded Tumor Tissues
title_fullStr Multiplex Immunofluorescence Tyramide Signal Amplification for Immune Cell Profiling of Paraffin-Embedded Tumor Tissues
title_full_unstemmed Multiplex Immunofluorescence Tyramide Signal Amplification for Immune Cell Profiling of Paraffin-Embedded Tumor Tissues
title_short Multiplex Immunofluorescence Tyramide Signal Amplification for Immune Cell Profiling of Paraffin-Embedded Tumor Tissues
title_sort multiplex immunofluorescence tyramide signal amplification for immune cell profiling of paraffin-embedded tumor tissues
topic Molecular Biosciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8118604/
https://www.ncbi.nlm.nih.gov/pubmed/33996912
http://dx.doi.org/10.3389/fmolb.2021.667067
work_keys_str_mv AT hernandezsharia multipleximmunofluorescencetyramidesignalamplificationforimmunecellprofilingofparaffinembeddedtumortissues
AT rojasfrank multipleximmunofluorescencetyramidesignalamplificationforimmunecellprofilingofparaffinembeddedtumortissues
AT laberianocaddie multipleximmunofluorescencetyramidesignalamplificationforimmunecellprofilingofparaffinembeddedtumortissues
AT lazcanorossana multipleximmunofluorescencetyramidesignalamplificationforimmunecellprofilingofparaffinembeddedtumortissues
AT wistubaignacio multipleximmunofluorescencetyramidesignalamplificationforimmunecellprofilingofparaffinembeddedtumortissues
AT parraedwinroger multipleximmunofluorescencetyramidesignalamplificationforimmunecellprofilingofparaffinembeddedtumortissues