Advanced NSCLC patients with high IL-6 levels have altered peripheral T cell population and signaling

doi:10.1016/j.lungcan.2019.03.014

Lung Cancer

Volume 131, May 2019, Pages 58-61

https://doi.org/10.1016/j.lungcan.2019.03.014 Get rights and content

Highlights

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Patients with high circulating IL-6 have altered T-Cell population distributions.
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Patients with high levels of circulating IL-6 have impaired STAT1 signaling.
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Impaired STAT1 signaling in T-Cells is produced in vitro by 4 day IL-6 treatment.

Abstract

Objectives

High levels of circulating interleukin-6 (IL-6) are associated with a poor prognosis in many types of cancer including non-small cell lung cancer (NSCLC). While the inflammatory cytokine can stimulate the immune system and promote tumor growth, it remains unclear how circulating IL-6 can potentiate a poor prognosis. We hypothesized that a mechanism for IL-6-associated poor prognosis is that these patients would have altered T-cell populations and impaired T-cell signaling.

Materials and methods

Plasma levels of IL-6 were measured using a Cytometric Bead Array. T-cell populations from Non-small cell lung cancer patients were characterized using surface markers by flow cytometry, and signaling in the T-cell populations were measured by PhosFlow cytometry.

Results

We determine that patients with high circulating IL-6 levels had distinct T cell characteristics relative to those with low levels. Patients with high levels of IL-6 had significantly more T_reg cells and elevated Programmed cell death protein-1 (PD-1) expression on CD4⁺, CD8⁺, Treg, and Th17 cells. These patients also showed impaired signal transducer and activator of transcription-1 (STAT1) signaling upon stimulation with IL-6 and phorbol 12-myristate 13-acetate (PMA), and T-Cells from a healthy donor that were treated for four days with IL-6 displayed a similar muting of STAT signaling, which verified the effect seen in patient samples.

Conclusions

This work directly links circulating IL-6 with other poor prognostic indicators, STAT1 and PD-1, and highlights the effects of circulating IL-6 on the immune system. Our data suggest that alteration in T cell populations and function may be a mechanism underlying the poor prognosis seen in NSCLC patients with high IL-6 levels.

Introduction

Modulation of the immune system through programmed cell death protein-1 (PD-1) inhibitors, i.e. nivolumab and pembrolizumab, can provide impressive clinical responses in non-small cell lung cancer (NSCLC) patients [1]. Blocking interactions between the inhibitory PD-1 on T cells with its ligand PD-L1 on the tumor or antigen presenting cells can reverse T-cell exhaustion and enhance tumor regression [2]. The striking results seen with checkpoint inhibitors highlights the importance of the immune system in treating cancer.

Aspects of the immune system can contribute to tumor growth and development. High levels of circulating IL-6 is associated with a poor prognosis in lung cancer patients [3]. IL-6 signaling is propagated through the STAT1/3 pathways [4]. Activation of the signal transducer and activator of transcription (STAT) and extracellular signal-regulator Kinase (ERK) pathways in peripheral blood cells can produce anti-tumor activity, and defective STAT1/3 signaling in peripheral blood T cells has been suggested as a poor prognostic indicator in breast cancer [5].

Although a variety of demographic features have been associated with high levels of circulating IL-6, i.e. age, smoking history, and chemotherapy, it remains unclear whether circulating IL-6 is a cause or a consequence of poor survival because possible mechanisms have not been elucidated [6]. Stromal and tumor-derived IL-6 can enhance the survival and proliferation of tumor cells, however, it is not known how circulating IL-6 could affect tumor populations. We proposed that a possible mechanism could be that circulating IL-6 may alter T cell populations or signaling to suppress the immune system and ultimately support tumor growth. We explored this possible relationship by characterizing T cell populations and signaling in advanced NSCLC patients with high and low circulating IL-6 levels.

Section snippets

Materials and methods

Patients who provided written informed consent to participate in an IRB-approved protocol were included in the study. Inclusion criteria included; stage IIIB-IV non-small cell lung cancer, beginning any line of therapy, measurable disease, and ability and willingness to sign an informed consent form. Peripheral blood mononuclear cells (PBMCs) from consented patients were isolated using CPT Cell Preparation Tubes (BD362761). PBMCs from a healthy donor were purchased from ZenBio (Research

Characterizing the effect of high plasma IL-6 on T cell populations

We measured the circulating IL-6 levels in 70 patients with advanced NSCLC. Patients with high levels of circulating IL-6 (greater than the median, 6.41 pg/ml) exhibited a shorter overall survival than those with low IL-6 levels, 215 versus 707 days (p < 0.001), respectively (Fig. S1). To better understand why circulating IL-6 has an adverse effect on survival, we characterized the PBMCs of patients with high and low IL-6 levels. We selected 23 patients with the highest and the 23 patients with

Discussion

Circulating IL-6 has frequently been associated with a poor prognosis in cancer patients, and while IL-6 has been linked to various demographic and behavioral characteristics, i.e. age, smoking history…, it remains unclear whether circulating IL-6 directly affects survival or if it is a consequence of poor survival [6]. We characterized T cell populations and signal in patients with high levels or circulating IL-6 and determined that these patient have altered features relative to those with

Contributions

SJR: Study design, data analysis, data interpretation and manuscript writing; XL and JZ: Study design, data collection, data interpretation and manuscript editing; BJ: Data collection and data analysis; HZ: Study design, data interpretation and manuscript editing; CPB: Study design, data interpretation, and manuscript editing.

Competing interests

None declared.

Ethics approval

Institutional Review Board of the Penn State College of Medicine.

Funding

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Acknowledgments

We thank Nate Sheaffer and Joseph Bednarczyk from the Penn State College of Medicine Flow Cytometry Core Facility for assistance with flow cytometry acquisition and analysis.

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View full text

Advanced NSCLC patients with high IL-6 levels have altered peripheral T cell population and signaling

Highlights

Abstract

Objectives

Materials and methods

Results

Conclusions

Introduction

Section snippets

Materials and methods

Characterizing the effect of high plasma IL-6 on T cell populations

Discussion

Contributions

Competing interests

Ethics approval

Funding

Acknowledgments

Nivolumab versus docetaxel in advanced nonsquamous non-small-cell lung Cancer

N. Engl. J. Med.

Targeting the PD-1/B7-H1(PD-L1) pathway to activate anti-tumor immunity

Curr. Opin. Immunol.

High systemic IL-6 is associated with worse prognosis in patients with non-small cell lung cancer

PLoS One

Targeting the IL-6/JAK/STAT3 signalling axis in cancer

Nat. Rev. Clin. Oncol.

IL6 signaling in peripheral blood t cells predicts clinical outcome in breast Cancer

Cancer Res.

Circulating interleukin-6 level is a prognostic marker for survival in advanced nonsmall cell lung cancer patients treated with chemotherapy

Int. J. Cancer