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Understanding immune phenotypes in human gastric disease tissues by multiplexed immunohistochemistry

BACKGROUND: Understanding immune phenotypes and human gastric disease in situ requires an approach that leverages multiplexed immunohistochemistry (mIHC) with multispectral imaging to facilitate precise image analyses. METHODS: We developed a novel 4-color mIHC assay based on tyramide signal amplifi...

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Autores principales: Ying, Le, Yan, Feng, Meng, Qiaohong, Yuan, Xiangliang, Yu, Liang, Williams, Bryan R. G., Chan, David W., Shi, Liyun, Tu, Yugang, Ni, Peihua, Wang, Xuefeng, Xu, Dakang, Hu, Yiqun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5639762/
https://www.ncbi.nlm.nih.gov/pubmed/29025424
http://dx.doi.org/10.1186/s12967-017-1311-8
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author Ying, Le
Yan, Feng
Meng, Qiaohong
Yuan, Xiangliang
Yu, Liang
Williams, Bryan R. G.
Chan, David W.
Shi, Liyun
Tu, Yugang
Ni, Peihua
Wang, Xuefeng
Xu, Dakang
Hu, Yiqun
author_facet Ying, Le
Yan, Feng
Meng, Qiaohong
Yuan, Xiangliang
Yu, Liang
Williams, Bryan R. G.
Chan, David W.
Shi, Liyun
Tu, Yugang
Ni, Peihua
Wang, Xuefeng
Xu, Dakang
Hu, Yiqun
author_sort Ying, Le
collection PubMed
description BACKGROUND: Understanding immune phenotypes and human gastric disease in situ requires an approach that leverages multiplexed immunohistochemistry (mIHC) with multispectral imaging to facilitate precise image analyses. METHODS: We developed a novel 4-color mIHC assay based on tyramide signal amplification that allowed us to reliably interrogate immunologic checkpoints, including programmed death-ligand 1 (PD-L1), cytotoxic T cells (CD8(+)T) and regulatory T cells (Foxp3), in formalin-fixed, paraffin-embedded tissues of various human gastric diseases. By observing cell phenotypes within the disease tissue microenvironment, we were able to determine specific co-localized staining combinations and various measures of cell density. RESULTS: We found that PD-L1 was expressed in gastric ulcer and in tumor cells (TCs), as well as in tumor-infiltrating immune cells (TIICs), but not in normal gastric mucosa or other gastric intraepithelial neoplastic tissues. Furthermore, we found no significant reduction in CD8(+)T cells, whereas the ratio of CD8(+)T:Foxp3 cells and CD8(+)T:PD-L1 cells was suppressed in tumor tissues and elevated in adjacent normal tissues. An unsupervised hierarchical analysis also identified correlations between CD8(+)T and Foxp3(+) tumor-infiltrating lymphocyte (TIL) densities and average PD-L1 levels. Three main groups were identified based on the results of CD8(+)T:PD-L1 ratios in gastric tumor tissues. Furthermore, integrating CD8(+)T:Foxp3 ratios, which increased the complexity for immune phenotype status, revealed 6–7 clusters that enabled the separation of gastric cancer patients at the same clinical stage into different risk-group subsets. CONCLUSIONS: Characterizing immune phenotypes in human gastric disease tissues via multiplexed immunohistochemistry may help guide PD-L1 clinical therapy. Observing unique disease tissue microenvironments can improve our understanding of immune phenotypes and cell interactions within these microenvironments, providing the ability to predict safe responses to immunotherapies. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12967-017-1311-8) contains supplementary material, which is available to authorized users.
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spelling pubmed-56397622017-10-18 Understanding immune phenotypes in human gastric disease tissues by multiplexed immunohistochemistry Ying, Le Yan, Feng Meng, Qiaohong Yuan, Xiangliang Yu, Liang Williams, Bryan R. G. Chan, David W. Shi, Liyun Tu, Yugang Ni, Peihua Wang, Xuefeng Xu, Dakang Hu, Yiqun J Transl Med Research BACKGROUND: Understanding immune phenotypes and human gastric disease in situ requires an approach that leverages multiplexed immunohistochemistry (mIHC) with multispectral imaging to facilitate precise image analyses. METHODS: We developed a novel 4-color mIHC assay based on tyramide signal amplification that allowed us to reliably interrogate immunologic checkpoints, including programmed death-ligand 1 (PD-L1), cytotoxic T cells (CD8(+)T) and regulatory T cells (Foxp3), in formalin-fixed, paraffin-embedded tissues of various human gastric diseases. By observing cell phenotypes within the disease tissue microenvironment, we were able to determine specific co-localized staining combinations and various measures of cell density. RESULTS: We found that PD-L1 was expressed in gastric ulcer and in tumor cells (TCs), as well as in tumor-infiltrating immune cells (TIICs), but not in normal gastric mucosa or other gastric intraepithelial neoplastic tissues. Furthermore, we found no significant reduction in CD8(+)T cells, whereas the ratio of CD8(+)T:Foxp3 cells and CD8(+)T:PD-L1 cells was suppressed in tumor tissues and elevated in adjacent normal tissues. An unsupervised hierarchical analysis also identified correlations between CD8(+)T and Foxp3(+) tumor-infiltrating lymphocyte (TIL) densities and average PD-L1 levels. Three main groups were identified based on the results of CD8(+)T:PD-L1 ratios in gastric tumor tissues. Furthermore, integrating CD8(+)T:Foxp3 ratios, which increased the complexity for immune phenotype status, revealed 6–7 clusters that enabled the separation of gastric cancer patients at the same clinical stage into different risk-group subsets. CONCLUSIONS: Characterizing immune phenotypes in human gastric disease tissues via multiplexed immunohistochemistry may help guide PD-L1 clinical therapy. Observing unique disease tissue microenvironments can improve our understanding of immune phenotypes and cell interactions within these microenvironments, providing the ability to predict safe responses to immunotherapies. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12967-017-1311-8) contains supplementary material, which is available to authorized users. BioMed Central 2017-10-12 /pmc/articles/PMC5639762/ /pubmed/29025424 http://dx.doi.org/10.1186/s12967-017-1311-8 Text en © The Author(s) 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Ying, Le
Yan, Feng
Meng, Qiaohong
Yuan, Xiangliang
Yu, Liang
Williams, Bryan R. G.
Chan, David W.
Shi, Liyun
Tu, Yugang
Ni, Peihua
Wang, Xuefeng
Xu, Dakang
Hu, Yiqun
Understanding immune phenotypes in human gastric disease tissues by multiplexed immunohistochemistry
title Understanding immune phenotypes in human gastric disease tissues by multiplexed immunohistochemistry
title_full Understanding immune phenotypes in human gastric disease tissues by multiplexed immunohistochemistry
title_fullStr Understanding immune phenotypes in human gastric disease tissues by multiplexed immunohistochemistry
title_full_unstemmed Understanding immune phenotypes in human gastric disease tissues by multiplexed immunohistochemistry
title_short Understanding immune phenotypes in human gastric disease tissues by multiplexed immunohistochemistry
title_sort understanding immune phenotypes in human gastric disease tissues by multiplexed immunohistochemistry
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5639762/
https://www.ncbi.nlm.nih.gov/pubmed/29025424
http://dx.doi.org/10.1186/s12967-017-1311-8
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