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Nondestructive, multiplex three-dimensional mapping of immune infiltrates in core needle biopsy

Steve Seung-Young Lee1,2, Vytautas P Bindokas3, Mark W Lingen4

  • 1Department of Molecular Genetics and Cell Biology, The University of Chicago, Chicago, IL, USA.

Laboratory Investigation; a Journal of Technical Methods and Pathology
|November 8, 2018
PubMed

Insights

Transparent tissue tomography (T3) enables 3D imaging of tumor immune infiltrates. This new tool reveals spatial immune cell distribution and relationships, offering a potential diagnostic for personalized cancer immunotherapy.

Area of Science:

  • Oncology
  • Immunology
  • Biomedical Imaging

Background:

  • Traditional tumor-infiltrating lymphocyte (TIL) enumeration lacks predictive power for immunotherapy.
  • Multiparametric flow cytometry loses spatial context, and multiplex immunohistochemistry analyzes single sections.
  • A need exists for tools that analyze immune infiltrates in 3D within the tumor microenvironment.

Purpose of the Study:

  • To introduce Transparent Tissue Tomography (T3) as a 3D imaging cytometry tool.
  • To demonstrate T3's capability for multiplexed immunofluorescent analysis in core needle biopsies.
  • To investigate the spatial distribution of cytotoxic T cells (CTLs) within the tumor microenvironment.

Main Methods:

  • Development and application of Transparent Tissue Tomography (T3) for 3D imaging cytometry.
  • Multiplexed immunofluorescent staining and analysis of core needle biopsies.
  • Image processing and machine learning for mapping CD3+CD8+ CTLs.
  • Analysis of Her2+ murine mammary tumors and human head and neck cancer specimens.

Main Results:

  • T3 revealed marked inhomogeneity in CD3+CD8+ CTL distribution within single needle cores.
  • A strong spatial correlation was found between CD3+CD8+ CTLs and microvasculature in EGFR+ parenchyma.
  • Significant patient-to-patient differences in immune cell distribution were observed in head and neck cancers.

Conclusions:

  • T3 provides rapid, 3D, quantitative maps of the tumor microenvironment and immune infiltrate.
  • T3 imaging cytometry offers a novel diagnostic approach for personalized cancer immunotherapy.
  • Understanding spatial immune cell distribution is crucial for predicting immunotherapy response.

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