High-dimensional multiplexed immunohistochemical characterization of immune contexture in human cancers

Grace Banik, Courtney B. Betts, Shannon M. Liudahl, Shamilene Sivagnanam, Rie Kawashima, Tiziana Cotechini, William Larson, Jeremy Goecks, Sara I. Pai, Daniel Clayburgh, Takahiro Tsujikawa, Lisa M. Coussens

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Biomarker assessments of tumor specimens is widely used in cancer research to audit tumor cell intrinsic as well as tumor cell extrinsic features including the diversity of immune, stromal, and mesenchymal cells. To comprehensively and quantitatively audit the tumor-immune microenvironment (TiME), we developed a novel multiplex immunohistochemistry (mIHC) platform and computational image processing workflow using a single formalin-fixed paraffin-embedded (FFPE) tissue section. Herein, we validated this platform using nine matched primary newly diagnosed and recurrent head and neck squamous cell carcinoma (HNSCC) sections sequentially subjected to immunodetection with a panel of 29 antibodies identifying malignant tumor cells, and 17 distinct leukocyte lineages and their functional states. Image cytometric analysis was applied to interpret chromogenic signals from digitally scanned and coregistered light microscopy-based images enabling identification and quantification of individual tumor cells, structural features, immune cell phenotypes and their functional state. In agreement with our previous study via a 12-plex imaging mIHC platform, myeloid-inflamed status in newly diagnosed primary tumors associated with significantly short progression free survival, independent of lymphoid-inflamed status. Spatial distribution of tumor and immune cell lineages in TiME was also examined and revealed statistically significant CD8+ T cell exclusion from tumor nests, whereas regulatory T cells and myeloid cells, when present in close proximity to tumor cells, highly associated with rapid cancer recurrence. These findings indicate presence of differential immune-spatial profiles in newly diagnosed and recurrent HNSCC, and establish the robustness of the 29-plex mIHC platform and associated analytics for quantitative analysis of single tissue sections revealing longitudinal TiME changes.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
PublisherAcademic Press Inc.
DOIs
StatePublished - Jan 1 2019

Publication series

NameMethods in Enzymology
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Fingerprint

Tumors
Neoplasms
Tumor Microenvironment
Cells
Immunohistochemistry
T-cells
Chromogenics
Tissue
Workflow
Cell Lineage
Regulatory T-Lymphocytes
Myeloid Cells
Tumor Biomarkers
Mesenchymal Stromal Cells
Paraffin
Formaldehyde
Disease-Free Survival
Microscopy
Image analysis
Spatial distribution

Keywords

  • Cancer immunology
  • Histopathology
  • Immunohistochemistry
  • Multiplex imaging

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Banik, G., Betts, C. B., Liudahl, S. M., Sivagnanam, S., Kawashima, R., Cotechini, T., ... Coussens, L. M. (2019). High-dimensional multiplexed immunohistochemical characterization of immune contexture in human cancers. In Methods in Enzymology (Methods in Enzymology). Academic Press Inc.. https://doi.org/10.1016/bs.mie.2019.05.039

High-dimensional multiplexed immunohistochemical characterization of immune contexture in human cancers. / Banik, Grace; Betts, Courtney B.; Liudahl, Shannon M.; Sivagnanam, Shamilene; Kawashima, Rie; Cotechini, Tiziana; Larson, William; Goecks, Jeremy; Pai, Sara I.; Clayburgh, Daniel; Tsujikawa, Takahiro; Coussens, Lisa M.

Methods in Enzymology. Academic Press Inc., 2019. (Methods in Enzymology).

Research output: Chapter in Book/Report/Conference proceedingChapter

Banik, G, Betts, CB, Liudahl, SM, Sivagnanam, S, Kawashima, R, Cotechini, T, Larson, W, Goecks, J, Pai, SI, Clayburgh, D, Tsujikawa, T & Coussens, LM 2019, High-dimensional multiplexed immunohistochemical characterization of immune contexture in human cancers. in Methods in Enzymology. Methods in Enzymology, Academic Press Inc. https://doi.org/10.1016/bs.mie.2019.05.039
Banik G, Betts CB, Liudahl SM, Sivagnanam S, Kawashima R, Cotechini T et al. High-dimensional multiplexed immunohistochemical characterization of immune contexture in human cancers. In Methods in Enzymology. Academic Press Inc. 2019. (Methods in Enzymology). https://doi.org/10.1016/bs.mie.2019.05.039
Banik, Grace ; Betts, Courtney B. ; Liudahl, Shannon M. ; Sivagnanam, Shamilene ; Kawashima, Rie ; Cotechini, Tiziana ; Larson, William ; Goecks, Jeremy ; Pai, Sara I. ; Clayburgh, Daniel ; Tsujikawa, Takahiro ; Coussens, Lisa M. / High-dimensional multiplexed immunohistochemical characterization of immune contexture in human cancers. Methods in Enzymology. Academic Press Inc., 2019. (Methods in Enzymology).
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