title: Quantitative Multiplex Immunohistochemistry Reveals Myeloid-Inflamed Tumor-Immune Complexity Associated with Poor Prognosis.
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.
authors.affiliation: 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.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Tsujikawa
authors.affiliation: Department of Cell, Developmental & Cancer Biology, Oregon Health and Science University, Portland, OR 97239, USA.
authors.forename: Sushil
authors.initials: S
authors.lastname: Kumar
authors.affiliation: Intel Health and Life Sciences, Intel Corporation, Hillsboro, OR 97124, USA.
authors.forename: Rohan N
authors.initials: RN
authors.lastname: Borkar
authors.affiliation: Intel Health and Life Sciences, Intel Corporation, Hillsboro, OR 97124, USA.
authors.forename: Vahid
authors.initials: V
authors.lastname: Azimi
authors.affiliation: Department of Biomedical Engineering, Oregon Health and Science University, Portland, OR 97239, USA.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Thibault
authors.affiliation: 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.
authors.forename: Young Hwan
authors.initials: YH
authors.lastname: Chang
authors.affiliation: Department of Computational Biology, Oregon Health and Science University, Portland, OR 97239, USA.
authors.forename: Ariel
authors.initials: A
authors.lastname: Balter
authors.affiliation: Department of Cell, Developmental & Cancer Biology, Oregon Health and Science University, Portland, OR 97239, USA.
authors.forename: Rie
authors.initials: R
authors.lastname: Kawashima
authors.affiliation: Department of Cell, Developmental & Cancer Biology, Oregon Health and Science University, Portland, OR 97239, USA.
authors.forename: Gina
authors.initials: G
authors.lastname: Choe
authors.affiliation: Department of Pathology, Oregon Health and Science University, Portland, OR 97239, USA.
authors.forename: David
authors.initials: D
authors.lastname: Sauer
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, Oregon Health and Science University, Portland, OR 97239, USA.
authors.forename: Edward
authors.initials: E
authors.lastname: El Rassi
authors.affiliation: 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.
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Clayburgh
authors.affiliation: 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.
authors.forename: Molly F
authors.initials: MF
authors.lastname: Kulesz-Martin
authors.affiliation: 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.
authors.forename: Eric R
authors.initials: ER
authors.lastname: Lutz
authors.affiliation: 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.
authors.forename: Lei
authors.initials: L
authors.lastname: Zheng
authors.affiliation: 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.
authors.forename: Elizabeth M
authors.initials: EM
authors.lastname: Jaffee
authors.affiliation: Department of Computational Biology, Oregon Health and Science University, Portland, OR 97239, USA.
authors.forename: Patrick
authors.initials: P
authors.lastname: Leyshock
authors.affiliation: 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.
authors.forename: Adam A
authors.initials: AA
authors.lastname: Margolin
authors.affiliation: 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.
authors.forename: Motomi
authors.initials: M
authors.lastname: Mori
authors.affiliation: 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.
authors.forename: Joe W
authors.initials: JW
authors.lastname: Gray
authors.affiliation: 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.
authors.forename: Paul W
authors.initials: PW
authors.lastname: Flint
authors.affiliation: 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.
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Coussens
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor / Q000032:analysis* / Q000378:metabolism; D002294:Carcinoma, Squamous Cell / Q000276:immunology*; D015331:Cohort Studies; D005260:Female; D006258:Head and Neck Neoplasms / Q000276:immunology*; D006801:Humans; D019044:Image Cytometry / Q000379:methods*; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D008297:Male; D008875:Middle Aged; D015166:Monitoring, Immunologic / Q000379:methods*; D011379:Prognosis; D018709:Statistics, Nonparametric; D046888:Tissue Array Analysis
chemical_list: D014408:Biomarkers, Tumor
keywords: cancer immunology; digital pathology; head and neck cancer; image cytometry; immunohistochemistry; multiplex; pancreatic cancer; tissue biomarker