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

• DOI: 10.1016/j.celrep.2017.03.037
• Publication Date: 2017-04-04T17:37:47Z
• Authors: 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-M...; Eric R. Lutz; Lei Zheng; Elizabeth M. Jaffee; Patrick Leyshock; Adam A. Margolin; Motomi Mori; Joe W. Gray; Paul W. Flint; Lisa M. Coussens
• Keywords: Male; 0301 basic medicine; QH301-705.5; pancreatic cancer; cancer immunology; Statistics, Nonparametric; Cohort Studies; 03 medical and health sciences; image cytometry; Monitoring, Immunologic; Biomarkers, Tumor; Image Processing, Computer-Assisted; Humans; Biology (General); Aged; Image Cytometry; Aged, 80 and over; tissue biomarker; Middle Aged; Prognosis; Immunohistochemistry; 3. Good health; multiplex; Head and Neck Neoplasms; Tissue Array Analysis; immunohistochemistry; Carcinoma, Squamous Cell; head and neck cancer; Female; digital pathology

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.