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Increased proportion of FoxP3+ regulatory T cells in tumor infiltrating lymphocytes is associated with tumor recurrence and reduced survival in patients with glioblastoma

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Abstract

Glioblastoma multiforme (GBM) is an aggressive malignancy associated with profound host immunosuppression mediated in part by FoxP3 expressing regulatory CD4+ T lymphocytes (Tregs) that down-regulate anti-tumor immunity. In order to assess whether FoxP3 was an independent driver differentially expressed in primary versus recurrent GBMs, we stained resected primary and recurrent GBM tumors for CD3, CD4, CD8 and FoxP3 expression using standard immunohistochemistry. Slides were scanned with a high-resolution scanner (ScanScope CS; Aperio), and image analysis software (Aperio ScanScope) was used to enumerate lymphocyte subpopulations allowing for high-throughput analysis and bypassing manual selection bias. As shown in previous studies, enumeration of individual lymphocyte populations did not correlate with clinical outcomes in patients with GBM. However, the CD4+ to regulatory FoxP3+ T cell ratio was diminished in recurrent disease, and increased CD3 and CD8+ to regulatory T cell ratios showed a positive correlation with survival outcomes in primary GBM. These results suggest that while absolute numbers of tumor infiltrating lymphocytes may not be informative for predicting clinical outcomes in patients with GBM, the effective balance of CD3, CD4 and CD8+ T cells to immunosuppressive FoxP3+ regulatory cells may influence clinical outcomes in this patient population.

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Abbreviations

GBM:

Glioblastoma multiforme

H&E:

Hematoxylin and eosin

IHC:

Immunohistochemistry

PBMCs:

Peripheral blood mononuclear cells

PPCv9:

Positive pixel count v9

Tregs:

Regulatory T lymphocytes

TILs:

Tumor infiltrating lymphocytes

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Acknowledgments

This work was supported by the National Institutes of Health (NIH)/National Cancer Institute/National Institute of Neurological Disorders and Stroke Grants [5R01-NS067037 (to D.A. Mitchell), 5R01-CA134844 (to D.A. Mitchell), 5P50-CA108786 (to J.H. Sampson/Bigner)] and a Clinical and Translational Science Awards Grant UL1RR024128 from the National Center for Research Resources, a component of the NIH and NIH Roadmap for Medical Research. Additional support was provided by Accelerate Brain Cancer Cure (ABC2), National Brain Tumor Society, the American Brain Tumor Association, and the Pediatric Brain Tumor Foundation of the United States.

Conflict of interest

The authors declare that they have no conflict of interest with the research presented in this paper.

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Authors and Affiliations

  1. University of Florida Brain Tumor Immunotherapy Program, Preston A. Wells, Jr. Center for Brain Tumor Therapy, McKnight Brain Institute, Department of Neurosurgery, University of Florida, 1149 South Newell Drive, P.O. Box 100265, Gainesville, FL, 32610, USA

    Elias J. Sayour, Gabriel De Leon & Duane A. Mitchell

  2. Department of Pathology, Duke University Medical Center, Durham, NC, 27710, USA

    Elias J. Sayour, Roger McLendon & John H. Sampson

  3. Duke Brain Tumor Immunotherapy Program, Division of Neurosurgery, Department of Surgery, Duke University Medical Center, Durham, NC, 27710, USA

    Pat McLendon, Renee Reynolds & John H. Sampson

  4. Department of Molecular Cancer Biology, Duke University Medical Center, Durham, NC, 27710, USA

    Gabriel De Leon

  5. Department of Pathology, University of Florida, Gainesville, FL, 32608, USA

    Jesse Kresak

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  1. Elias J. Sayour
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  2. Pat McLendon
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  3. Roger McLendon
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Correspondence to Duane A. Mitchell.

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Elias J. Sayour and Pat McLendon have contributed equally to this work.

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Sayour, E.J., McLendon, P., McLendon, R. et al. Increased proportion of FoxP3+ regulatory T cells in tumor infiltrating lymphocytes is associated with tumor recurrence and reduced survival in patients with glioblastoma. Cancer Immunol Immunother 64, 419–427 (2015). https://doi.org/10.1007/s00262-014-1651-7

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