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Heterogeneity of CD8+ tumor-infiltrating lymphocytes in non-small-cell lung cancer: impact on patient prognostic assessments and comparison of quantification by different sampling strategies

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Abstract

Introduction

Infiltration of non-small-cell lung cancer (NSCLC) by CD8+ T lymphocytes predicts improved patient survival; however, heterogeneity of intratumoral localization complicates this assessment. Strategies for tumor sampling may not accurately represent the whole tumor. We hypothesized that sampling strategies may alter the identification of tumors with high CD8 density and affect the prognostic significance.

Patients and methods

Twenty-three primary NSCLC tumors were immunohistochemically stained for CD8 and were assessed using automated software with eight different sampling strategies or the whole tumor. Results of all sampling strategies were compared to the whole tumor counts (paired t tests, Pearson’s r). Associations between CD8 densities and overall survival were assessed (log-rank test).

Results

Counts from all eight sampling strategies significantly correlated with whole tumor counts (p ≤ 0.001). However, the magnitude of CD8+ cell counts and categorization into high vs low infiltrate groups were affected by the sampling strategy. The most concordant values were derived from random sampling of 20 % of the tumor, a simulated core biopsy, or from sampling the tumor center. TIL infiltration was associated with survival when sampling the center (p = 0.038), but not the invasive margin (p > 0.2) or other strategies.

Conclusion

Different tumor sampling strategies may yield discordant TIL density results and different stratification for risk assessment. Small biopsies may be particularly unrepresentative. Random sampling of larger tumor areas is recommended. Enumerating CD8+ T cells in the tumor center may have prognostic value.

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Abbreviations

CTS:

Central tumor sampling

DLA:

Dense lymphoid aggregate

MAX:

Largest density

MIN:

Smallest density

NSCLC:

Non-small-cell lung cancer

STDEV:

Standard deviation

TMA:

Tissue microarray

TME:

Tumor microenvironment

WT:

Whole tumor

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Acknowledgments

The authors thank Dr. David R. Jones for partially supporting this work through the David R. Jones Lung Cancer Fund.

Funding

This work has been supported in part by the National Cancer Institute: T32 CA163177 (Joseph Obeid and Yinin Hu), K25 CA181638 (Nolan Wages), and U01 CA178846 (Craig Slingluff Jr.).

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Authors and Affiliations

  1. Department of Surgery, University of Virginia, Charlottesville, VA, USA

    Joseph M. Obeid, Yinin Hu, Donna H. Deacon & Craig L. Slingluff Jr.

  2. Department of Public Health Sciences, University of Virginia, Charlottesville, VA, USA

    Nolan A. Wages

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  1. Joseph M. Obeid
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Correspondence to Craig L. Slingluff Jr..

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Conflict of interest

Craig Slingluff Jr., received consulting fees from Immatics, Curetech, Castle Biosciences, and Polynoma, has royalties in University of Virginia Licensing and Ventures Group and was provided PD-1 antibody for use in investigator initiated trial from Merck. All other authors have no conflict of interest to disclose.

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Obeid, J.M., Wages, N.A., Hu, Y. et al. Heterogeneity of CD8+ tumor-infiltrating lymphocytes in non-small-cell lung cancer: impact on patient prognostic assessments and comparison of quantification by different sampling strategies. Cancer Immunol Immunother 66, 33–43 (2017). https://doi.org/10.1007/s00262-016-1908-4

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