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1. Quantitative Multiplex Immunohistochemistry Reveals Myeloid-Inflamed Tumor-Immune Complexity Associated with Poor Prognosis

Author

Takahiro Tsujikawa, Sushil Kumar, Rohan N. Borkar, Vahid Azimi, Guillaume Thibault, Young Hwan Chang, Ariel Balter, Rie Kawashima, Gina Choe, David Sauer, Edward El Rassi, Daniel R. Clayburgh, Molly F. Kulesz-Martin, Eric R. Lutz, Lei Zheng, Elizabeth M. Jaffee, Patrick Leyshock, Adam A. Margolin, Motomi Mori, Joe W. Gray, Paul W. Flint, and Lisa M. Coussens

Subjects
cancer immunology, multiplex, immunohistochemistry, digital pathology, image cytometry, head and neck cancer, pancreatic cancer, tissue biomarker, Biology (General), QH301-705.5
Abstract

Here, we describe a multiplexed immunohistochemical platform with computational image processing workflows, including image cytometry, enabling simultaneous evaluation of 12 biomarkers in one formalin-fixed paraffin-embedded tissue section. To validate this platform, we used tissue microarrays containing 38 archival head and neck squamous cell carcinomas and revealed differential immune profiles based on lymphoid and myeloid cell densities, correlating with human papilloma virus status and prognosis. Based on these results, we investigated 24 pancreatic ductal adenocarcinomas from patients who received neoadjuvant GVAX vaccination and revealed that response to therapy correlated with degree of mono-myelocytic cell density and percentages of CD8+ T cells expressing T cell exhaustion markers. These data highlight the utility of in situ immune monitoring for patient stratification and provide digital image processing pipelines to the community for examining immune complexity in precious tissue sections, where phenotype and tissue architecture are preserved to improve biomarker discovery and assessment.

Published
2017
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2. Quantitative Multiplex Immunohistochemistry Reveals Myeloid-Inflamed Tumor-Immune Complexity Associated with Poor Prognosis

Author

Motomi Mori, Takahiro Tsujikawa, Molly Kulesz-Martin, Lei Zheng, Eric R. Lutz, Paul W. Flint, Daniel R. Clayburgh, Patrick Leyshock, Vahid Azimi, Adam A. Margolin, Rie Kawashima, Young Hwan Chang, Elizabeth M. Jaffee, Gina Choe, Joe W. Gray, Ariel Balter, Guillaume Thibault, Sushil Kumar, David Sauer, Rohan N. Borkar, Edward El Rassi, and Lisa M. Coussens

Subjects
0301 basic medicine, Male, Myeloid, pancreatic cancer, Biology, cancer immunology, General Biochemistry, Genetics and Molecular Biology, Statistics, Nonparametric, Article, Cohort Studies, 03 medical and health sciences, Immune system, Monitoring, Immunologic, Pancreatic cancer, medicine, Biomarkers, Tumor, Image Processing, Computer-Assisted, Humans, Biomarker discovery, lcsh:QH301-705.5, Cancer immunology, Aged, Image Cytometry, Aged, 80 and over, Tissue microarray, tissue biomarker, Middle Aged, medicine.disease, Prognosis, GVAX, Immunohistochemistry, 3. Good health, multiplex, 030104 developmental biology, medicine.anatomical_structure, lcsh:Biology (General), Head and Neck Neoplasms, Tissue Array Analysis, Immunology, Cancer research, Carcinoma, Squamous Cell, head and neck cancer, Female, digital pathology
Abstract

Here we describe a multiplexed immunohistochemical platform, with computational image processing workflows including image cytometry, enabling simultaneous evaluation of 12 biomarkers in one formalin-fixed paraffin-embedded tissue section. To validate this platform, we used tissue microarrays containing 38 archival head and neck squamous cell carcinomas, and revealed differential immune profiles based on lymphoid and myeloid cell densities, correlating with human papilloma virus status and prognosis. Based on these results, we investigated 24 pancreatic ductal adenocarcinomas from patients who received neoadjuvant GVAX vaccination, and revealed that response to therapy correlated with degree of mono-myelocytic cell density, and percentages of CD8+ T cells expressing T cell exhaustion markers. These data highlight the utility of in situ immune monitoring for patient stratification, and provide digital image processing pipelines (https://github.com/multiplexIHC/cppipe) to the community for examining immune complexity in precious tissue sections, where phenotype and tissue architecture are preserved to thus improve biomarker discovery and assessment.

Published
2016

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