Nondestructive, multiplex three-dimensional mapping of immune infiltrates in core needle biopsy.
IF: 5.502
Cited by: 9


Enumeration of tumor-infiltrating lymphocytes (TILs) in H&E stained tissue sections has demonstrated limited value in predicting immune responses to cancer immunotherapy, likely reflecting the diversity of cell types and immune activation states among tumor infiltrates. Multiparametric flow cytometry enables robust phenotypic and functional analysis to distinguish suppression from activation, but tissue dissociation eliminates spatial context. Multiplex methods for immunohistochemistry (IHC) are emerging, but these interrogate only a single tissue section at a time. Here, we report transparent tissue tomography (T3) as a tool for three-dimensional (3D) imaging cytometry in the complex architecture of the tumor microenvironment, demonstrating multiplexed immunofluorescent analysis in core needle biopsies. Using T3 imaging, image processing and machine learning to map CD3+CD8+ cytotoxic T cells (CTLs) in whole core needle biopsies from Her2+ murine mammary tumors and human head and neck surgical specimens revealed marked inhomogeneity within single needle cores, confirmed by serial section IHC. Applying T3 imaging cytometry, we discovered a strong spatial correlation between CD3+CD8+ CTLs and microvasculature in the EGFR+ parenchyma, revealing significant differences among head and neck cancer patients. These results show that T3 offers simple and rapid access to three-dimensional and quantitative maps of the tumor microenvironment and immune infiltrate, offering a new diagnostic tool for personalized cancer immunotherapy.



MeSH terms

Aged, 80 and over
Biopsy, Large-Core Needle
CD8-Positive T-Lymphocytes
Head and Neck Neoplasms
Image Cytometry
Imaging, Three-Dimensional
Lymphocytes, Tumor-Infiltrating
Mammary Neoplasms, Animal
Mice, Inbred BALB C
Mice, Transgenic
Middle Aged
Reproducibility of Results
T-Lymphocytes, Cytotoxic
Tomography, X-Ray Computed


Lee, Steve Seung-Young
Bindokas, Vytautas P
Lingen, Mark W
Kron, Stephen J

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