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Preferential expansion of pro-inflammatory Tregs in human non-small cell lung cancer

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Cancer Immunology, Immunotherapy Aims and scope Submit manuscript

Abstract

Objectives

Lung cancer is the leading cause of cancer-related death in the USA. Regulatory T cells (Tregs) normally function to temper immune responses and decrease inflammation. Previous research has demonstrated different subsets of Tregs with contrasting anti- or pro-inflammatory properties. This study aimed to determine Treg subset distributions and characteristics present in non-small cell lung cancer (NSCLC) patients.

Methods

Peripheral blood was collected from healthy controls (HC) and NSCLC patients preceding surgical resection, and mononuclear cells were isolated, stained, and analyzed by flow cytometry. Tregs were defined by expression of CD4 and CD25 and classified into CD45RA+Foxp3int (naïve, Fr. I) or CD45RA−Foxp3hi (activated Fr. II). Activated conventional T cells were CD4+CD45RA−Foxp3int (Fr. III).

Results

Samples from 23 HC and 26 NSCLC patients were collected. Tregs isolated from patients with NSCLC were found to have enhanced suppressive function on naive T cells. Cancer patients had significantly increased frequencies of activated Tregs (fraction II: FrII), 17.5 versus 3.2 % (P < 0.001). FrII Tregs demonstrated increased RORγt and IL17 expression and decreased IL10 expression compared to Tregs from HC, indicating pro-inflammatory characteristics.

Conclusions

This study demonstrates that a novel subset of Tregs with pro-inflammatory characteristics preferentially expand in NSCLC patients. This Treg subset appears identical to previously reported pro-inflammatory Tregs in human colon cancer patients and in mouse models of polyposis. We expect the pro-inflammatory Tregs in lung cancer to contribute to the immune pathogenesis of disease and propose that targeting this Treg subset may have protective benefits in NSCLC.

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Abbreviations

ADC:

Adenocarcinoma

CC:

Colon cancer

CFSE:

Carboxyfluorescein succinimidyl ester

FACS:

Fluorescence-activated cell sorting

Foxp3:

Forkhead box P3

Fr:

Fraction

HC:

Healthy control

IL-10:

Interleukin 10

IL-17:

Interleukin 17

MC:

Mast cells

MNC:

Mononuclear cells

NSCLC:

Non-small cell lung cancer

PB:

Peripheral blood

PBMC:

Peripheral blood mononuclear cells

RORγt:

Retinoic acid receptor related orphan receptor gamma T

SCC:

Squamous cell cancer

TH17:

T helper 17 cells

Tregs:

T regulatory cells

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Acknowledgments

David Bentrem is supported by a Career Development Award from the Health Services Research and Development Service of the Department of Veterans Affairs. Khashayarsha Khazaie is supported by National Institutes of Health (NIH)/National Cancer Institute (NCI) Grant 1R01CA160436 and NIH/NIAID R01 AI108682-01. Fotini Gounari is supported by National Institutes of Health (NIH)/National Institute of Allergy and Infectious Diseases (NIAID) R01 AI108682-01. We would like to acknowledge Dr. Alberto de Hoyos for his contribution in obtaining lung cancer tissue.

Conflict of interest

The authors have no disclosures of conflict of interest.

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Author notes

    Authors and Affiliations

    1. Department of Surgery, Feinberg School of Medicine, Northwestern University, Suite 650, 676 N St. Clair, Chicago, IL, 60611, USA

      Joseph D. Phillips, Lawrence M. Knab, Leila Haghi, Malcolm M. DeCamp, Shari L. Meyerson & David J. Bentrem

    2. Robert H. Lurie Comprehensive Cancer Center, Northwestern University, Chicago, IL, USA

      Joseph D. Phillips, Lawrence M. Knab, Nichole R. Blatner, Leila Haghi, Michael J. Heiferman, Jeffrey R. Heiferman & David J. Bentrem

    3. Jesse Brown VA Medical Center, Chicago, IL, USA

      Shari L. Meyerson & David J. Bentrem

    4. Department of Medicine, Section of Rheumatology, University of Chicago, Chicago, IL, USA

      Fotini Gounari

    5. Mayo Clinic College of Medicine, Rochester, MN, USA

      Khashayarsha Khazaie

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    1. Joseph D. Phillips
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    Correspondence to Lawrence M. Knab.

    Additional information

    Joseph D. Phillips and Lawrence M. Knab have contributed equally to this work.

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    Phillips, J.D., Knab, L.M., Blatner, N.R. et al. Preferential expansion of pro-inflammatory Tregs in human non-small cell lung cancer. Cancer Immunol Immunother 64, 1185–1191 (2015). https://doi.org/10.1007/s00262-015-1725-1

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    • DOI: https://doi.org/10.1007/s00262-015-1725-1

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