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.
Author Info: (1) Department of Cell, Developmental & Cancer Biology, Oregon Health and Science University, Portland, OR 97239, USA; Department of Otolaryngology-Head and Neck Surgery, Oregon He
Author Info: (1) Department of Cell, Developmental & Cancer Biology, Oregon Health and Science University, Portland, OR 97239, USA; Department of Otolaryngology-Head and Neck Surgery, Oregon Health and Science University, Portland, OR 97239, USA. (2) Department of Cell, Developmental & Cancer Biology, Oregon Health and Science University, Portland, OR 97239, USA. (3) Intel Health and Life Sciences, Intel Corporation, Hillsboro, OR 97124, USA. (4) Intel Health and Life Sciences, Intel Corporation, Hillsboro, OR 97124, USA. (5) Department of Biomedical Engineering, Oregon Health and Science University, Portland, OR 97239, USA. (6) Department of Biomedical Engineering, Oregon Health and Science University, Portland, OR 97239, USA; Department of Computational Biology, Oregon Health and Science University, Portland, OR 97239, USA. (7) Department of Computational Biology, Oregon Health and Science University, Portland, OR 97239, USA. (8) Department of Cell, Developmental & Cancer Biology, Oregon Health and Science University, Portland, OR 97239, USA. (9) Department of Cell, Developmental & Cancer Biology, Oregon Health and Science University, Portland, OR 97239, USA. (10) Department of Pathology, Oregon Health and Science University, Portland, OR 97239, USA. (11) Department of Otolaryngology-Head and Neck Surgery, Oregon Health and Science University, Portland, OR 97239, USA. (12) Department of Otolaryngology-Head and Neck Surgery, Oregon Health and Science University, Portland, OR 97239, USA; Knight Cancer Institute, Oregon Health and Science University, Portland, OR 97239, USA. (13) Department of Cell, Developmental & Cancer Biology, Oregon Health and Science University, Portland, OR 97239, USA; Department of Dermatology, Oregon Health and Science University, Portland, OR 97239, USA; Knight Cancer Institute, Oregon Health and Science University, Portland, OR 97239, USA. (14) Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; The Sidney Kimmel Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; The Skip Viragh Center for Pancreatic Cancer Research and Clinical Care, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. (15) Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Surgery, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; The Sidney Kimmel Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; The Skip Viragh Center for Pancreatic Cancer Research and Clinical Care, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. (16) Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; The Sidney Kimmel Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; The Skip Viragh Center for Pancreatic Cancer Research and Clinical Care, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. (17) Department of Computational Biology, Oregon Health and Science University, Portland, OR 97239, USA. (18) Department of Computational Biology, Oregon Health and Science University, Portland, OR 97239, USA; OHSU Center for Spatial Systems Biomedicine, Oregon Health and Science University, Portland, OR 97239, USA; Knight Cancer Institute, Oregon Health and Science University, Portland, OR 97239, USA. (19) School of Public Health, Oregon Health and Science University, Portland, OR 97239, USA; Knight Cancer Institute, Oregon Health and Science University, Portland, OR 97239, USA. (20) Department of Biomedical Engineering, Oregon Health and Science University, Portland, OR 97239, USA; OHSU Center for Spatial Systems Biomedicine, Oregon Health and Science University, Portland, OR 97239, USA; Knight Cancer Institute, Oregon Health and Science University, Portland, OR 97239, USA. (21) Department of Otolaryngology-Head and Neck Surgery, Oregon Health and Science University, Portland, OR 97239, USA; Knight Cancer Institute, Oregon Health and Science University, Portland, OR 97239, USA. (22) Department of Cell, Developmental & Cancer Biology, Oregon Health and Science University, Portland, OR 97239, USA; Knight Cancer Institute, Oregon Health and Science University, Portland, OR 97239, USA. Electronic address: coussenl@ohsu.edu.
