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Interferon beta increases NK cell cytotoxicity against tumor cells in patients with nasopharyngeal carcinoma via tumor necrosis factor apoptosis-inducing ligand

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Abstract

Background

Nasopharyngeal carcinoma (NPC) is an EBV-associated neoplasm occurring endemically in Southeast Asia and sporadically all over the world. In children and adolescents, high cure rates have been obtained using chemotherapy, radiochemotherapy and maintenance therapy with interferon beta (IFNβ). The mechanism by which IFNβ contributes to a low systemic relapse rate has not yet been fully revealed.

Patients and methods

NK cells and serum samples from two patients with NPC were analyzed before and at different time points during IFNβ therapy, for assessment of TRAIL expression and NK cell cytotoxicity. Cytotoxicity was measured using the calcein release assay and the contribution of different death effector pathways was analyzed using specific inhibitors.

Results

Treatment with IFNβ induced TRAIL expression on patients’ NK cells and increased their cytotoxicity against NPC targets in vitro. NK cell-mediated cytotoxicity was predominately mediated via TRAIL. IFNβ also induced the production of soluble TRAIL (sTRAIL) by NK cells and its release upon contact with NPC cells. IFNβ treatment increased serum levels of sTRAIL in patients. Moreover, sTRAIL concentrated from patients’ serum samples induced apoptosis ex vivo in NPC cells from a patient-derived xenograft.

Conclusion

Increased cytotoxicity of NK cells against NPC cells and increased serum levels of biologically active TRAIL in patients treated with IFNβ could be a means to eliminate micrometastatic disease and explain the low systemic relapse rate in this patient group.

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Abbreviations

Calcein-AM:

Calcein-acetyoxymethyl

EBV:

Epstein–Barr virus

FAS:

First apoptosis signal

FASL:

FAS ligand

GPOH:

German Society of Pediatric Oncology and Hematology

IFNα, -β:

Interferon alpha, -beta

IFNAR1, -2:

Interferon alpha and beta receptor subunit 1, -2

NGS:

Next-generation sequencing

NPC:

Nasopharyngeal carcinoma

PDX:

Patient-derived xenograft

RFU:

Relative fluorescence units

sTRAIL:

Soluble TRAIL

TRAIL-R1, -R2:

TRAIL receptor 1, -2

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Acknowledgements

We thank Anshu Babbar who carefully proofread the manuscript.

Funding

The study has been funded internally by the Medical Faculty, Rhenish-Westphalian Technical University Aachen, Germany.

Author information

Authors and Affiliations

  1. Division of Pediatric Hematology, Oncology and Stem Cell Transplantation, Medical Faculty, Rhenish-Westphalian Technical University, Pauwelsstraße 30, 52074, Aachen, Germany

    Anna Makowska, Sabrina Franzen, Lian Shen & Udo Kontny

  2. Institute of Pathology, Medical Faculty, Rhenish-Westphalian Technical University, Aachen, Germany

    Till Braunschweig

  3. Interdisciplinary Center for Clinical Research, Medical Faculty, Rhenish-Westphalian Technical University, Aachen, Germany

    Bernd Denecke

  4. CNRS, UMR 8126, Gustave Roussy, Université Paris-Saclay, 94805, Villejuif, France

    Valentin Baloche & Pierre Busson

Authors
  1. Anna Makowska
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  2. Sabrina Franzen
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  3. Till Braunschweig
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  5. Lian Shen
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Contributions

Conception and design: AM, UK; development of methodology: AM, BD, VB, PB; acquisition of data: AM, SF, TB, LS, BD; analysis and interpretation of data: AM, TB, BD, PB, UK; writing and review of the manuscript: AM, PB, UK; material support: BD, PB; study supervision: UK.

Corresponding author

Correspondence to Udo Kontny.

Ethics declarations

Conflict of interest

The authors declare that they have no conflicts of interest.

Ethical standards

The final protocol was approved by the ethics committee of the Rhenish-Westphalian Technical University, Aachen, Germany [EK 005/18]. The study was conducted in accordance with the Declaration of Helsinki (2013 revision).

Ethical approval

Procedures for mouse handling and xenografts were reviewed and approved by the Ethics Committee for animal experimentation n°26 (Gustave Roussy, Villejuif, France), in accordance with the European directive 2010/63/EU and the decrees of the French ministry of Agriculture R. 214-87 to R. 214-126. The approval was given in November 26, 2015, under the Number Apafis #1605—2015090216498538-v2.

Informed consent

Written informed consent was obtained from all individual participants included in the study. Patients were treated at the Uniklinik RWTH Aachen. Informed consent including use of biological specimen (tumor, peripheral blood, urine) and data acquisition and processing was obtained from patients prior to the initiation of the study. Informed consent including the use of peripheral blood and anonymous processing of data was obtained from healthy volunteers who were part of the laboratory staff.

Animal source

Swiss nude mice were bred in the animal facility at Gustave Roussy.

Cell line authentication

C666-1 was a gift from Prof. Fei–Fei Liu, University of Toronto, Canada [49] and the SV40T-antigen immortalized nasopharyngeal epithelial cell line NP69 [50] was obtained from Prof. George Tsao (The Chinese University of Hong Kong, Hong Kong, China). Cell authentication was done using short tandem repeated profiles as described previously [13], and cell lines were tested at regular intervals by PCR to rule out mycoplasma contamination. Authentication of C17 cell was done by checking of HLA class I alleles by PCR (A02.01/A26.01–B44.02/B51.01).

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Makowska, A., Franzen, S., Braunschweig, T. et al. Interferon beta increases NK cell cytotoxicity against tumor cells in patients with nasopharyngeal carcinoma via tumor necrosis factor apoptosis-inducing ligand. Cancer Immunol Immunother 68, 1317–1329 (2019). https://doi.org/10.1007/s00262-019-02368-y

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