Abstract
BackgroundTo date, pathological examination of specimens remains largely qualitative. Quantitative measures of tissue spatial features are generally not captured. To gain additional mechanistic and prognostic insights, a need for quantitative architectural analysis arises in studying immune cell-cancer interactions within the tumor microenvironment and tumor-draining lymph nodes (TDLNs).Methodology/Principal FindingsWe present a novel, quantitative image analysis approach incorporating 1) multi-color tissue staining, 2) high-resolution, automated whole-section imaging, 3) custom image analysis software that identifies cell types and locations, and 4) spatial statistical analysis. As a proof of concept, we applied this approach to study the architectural patterns of T and B cells within tumor-draining lymph nodes from breast cancer patients versus healthy lymph nodes. We found that the spatial grouping patterns of T and B cells differed between healthy and breast cancer lymph nodes, and this could be attributed to the lack of B cell localization in the extrafollicular region of the TDLNs.Conclusions/SignificanceOur integrative approach has made quantitative analysis of complex visual data possible. Our results highlight spatial alterations of immune cells within lymph nodes from breast cancer patients as an independent variable from numerical changes. This opens up new areas of investigations in research and medicine. Future application of this approach will lead to a better understanding of immune changes in the tumor microenvironment and TDLNs, and how they affect clinical outcomes.
Highlights
Medicine is evolving from a qualitative to a quantitative science
Quantitative image analysis approach To analyze the spatial relationships between several different cell populations within tissue, samples were stained using multiple chromogens and the entire tissue sections were imaged at high resolution
Estrogen and progesterone (ER/PR) receptor expression is generally described by roughly estimating the intensity and percentage of tumor cells staining, such as 3+ and 30%
Summary
Medicine is evolving from a qualitative to a quantitative science This has led to improved diagnostic and prognostic tools as well as novel therapies. Qualitative measures of tumor characteristics remain the standard of practice. Spatial measures, such as average distance between cells or local density of cells, are generally overlooked in research or clinical studies, largely due to the laborious nature of manually scoring histological data, generating a quantitative code which captures the histological characteristics, and performing statistical analysis of this data. Quantitative measures of tissue spatial features are generally not captured. To gain additional mechanistic and prognostic insights, a need for quantitative architectural analysis arises in studying immune cell-cancer interactions within the tumor microenvironment and tumordraining lymph nodes (TDLNs)
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