CD8+ T cell infiltration in breast and colon cancer: A histologic and statistical analysis

PLoS One. 2018 Jan 10;13(1):e0190158. doi: 10.1371/journal.pone.0190158. eCollection 2018.

Abstract

The prevalence of cytotoxic tumor infiltrating lymphocytes (TILs) has demonstrated prognostic value in multiple tumor types. In particular, CD8 counts (in combination with CD3 and CD45RO) have been shown to be superior to traditional UICC staging in colon cancer patients and higher total CD8 counts have been associated with better survival in breast cancer patients. However, immune infiltrate heterogeneity can lead to potentially significant misrepresentations of marker prevalence in routine histologic sections. We examined step sections of breast and colorectal cancer samples for CD8+ T cell prevalence by standard chromogenic immunohistochemistry to determine marker variability and inform practice of T cell biomarker assessment in formalin-fixed, paraffin-embedded (FFPE) tissue samples. Stained sections were digitally imaged and CD8+ lymphocytes within defined regions of interest (ROI) including the tumor and surrounding stroma were enumerated. Statistical analyses of CD8+ cell count variability using a linear model/ANOVA framework between patients as well as between levels within a patient sample were performed. Our results show that CD8+ T-cell distribution is highly homogeneous within a standard tissue sample in both colorectal and breast carcinomas. As such, cytotoxic T cell prevalence by immunohistochemistry on a single level or even from a subsample of biopsy fragments taken from that level can be considered representative of cytotoxic T cell infiltration for the entire tumor section within the block. These findings support the technical validity of biomarker strategies relying on CD8 immunohistochemistry.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Biopsy / methods
  • Breast Neoplasms / immunology*
  • Breast Neoplasms / pathology
  • CD8-Positive T-Lymphocytes / immunology
  • CD8-Positive T-Lymphocytes / pathology*
  • Carcinoma / immunology*
  • Carcinoma / pathology
  • Colorectal Neoplasms / immunology*
  • Colorectal Neoplasms / pathology
  • Computer Simulation
  • Cytotoxicity, Immunologic
  • Female
  • Histological Techniques
  • Humans
  • Image Processing, Computer-Assisted
  • Lymphocyte Count
  • Lymphocytes, Tumor-Infiltrating / immunology
  • Lymphocytes, Tumor-Infiltrating / pathology*
  • Male
  • Middle Aged
  • Prognosis
  • ROC Curve
  • T-Lymphocyte Subsets / immunology
  • T-Lymphocyte Subsets / pathology*
  • T-Lymphocytes, Cytotoxic / immunology
  • T-Lymphocytes, Cytotoxic / pathology
  • Tumor Microenvironment

Grants and funding

We also confirm here that while all funding sources for this study were internal to Genentech, Inc., the funders had no role in study design, data collection and analysis, decision to publish, orpreparation of the manuscript.