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Stimulation of chemokines in human endometrial stromal cells by tumor necrosis factor-α and interferon-γ is similar under apoptotic and non-apoptotic conditions

  • Gynecologic Endocrinology and Reproductive Medicine
  • Published:
Archives of Gynecology and Obstetrics Aims and scope Submit manuscript

Abstract

Purpose

Tumor necrosis factor (TNF)-α and interferon (IFN)-γ are pro-inflammatory cytokines which have been shown to be involved in the pathophysiology of implantation disorders. Both cytokines in combination are able to sensitize primarily resistant human endometrial stromal cells (ESCs) to Fas-induced apoptosis. Since CCL (CC-chemokine ligand) 5 and CCL2 are important regulators of the endometrial immune cell population, we examined the impact of TNF-α and IFN-γ on these two chemokines under non-apoptotic and apoptotic conditions.

Methods

ESCs were isolated from hysterectomy specimens, decidualized in vitro and incubated with TNF-α, IFN-γ, an activating anti-Fas antibody and a caspase-inhibitor. CCL5 and CCL2 were measured using ELISA and real-time RT-PCR. Apoptosis was determined by flow cytometry, and cellular viability and membrane integrity were measured by fluorescent assays.

Results

The secretion of CCL5 and CCL2 was stimulated in undifferentiated and decidualized ESCs by the combination of TNF-α and IFN-γ under non-apoptotic as well as apoptotic (with Fas-stimulation in parallel) conditions. TNF-α or IFN-γ alone did not have this effect. The stimulatory influence of TNF-α plus IFN-γ on CCL5 and CCL2 in ESCs was also seen on the transcriptional level. Inhibition of cell death by a caspase-inhibitor had no influence on the secretion of CCL5 and CCL2 in ESCs under apoptotic stimulation.

Conclusion

TNF-α and IFN-γ modulate the secretion of chemokines in ESCs independently of Fas-induced apoptosis. These results suggest a constant response pattern on pro-inflammatory cytokines within the population of human ESCs.

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology, University of Heidelberg, Im Neuenheimer Feld 440, 69120, Heidelberg, Germany

    Julia Spratte, Florian Schütz, Joachim Rom & Herbert Fluhr

  2. Department of Obstetrics and Gynecology, University of Lübeck, Ratzeburger Allee 160, 23538, Lübeck, Germany

    Henriette Princk

  3. Department of Obstetrics and Gynecology, University of Greifswald, Sauerbruchstraße, 17487, Greifswald, Germany

    Marek Zygmunt

Authors
  1. Julia Spratte
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  2. Henriette Princk
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  4. Joachim Rom
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  5. Marek Zygmunt
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  6. Herbert Fluhr
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Contributions

JS and HP designed and performed experiments, analyzed the results and revised the manuscript. JR and FS contributed to the interpretation of the results and the writing of the manuscript. MZ supervised this study and revised the manuscript. HF initiated and coordinated this study, interpreted the data and wrote the manuscript.

Corresponding author

Correspondence to Julia Spratte.

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Funding

This study was funded by a grant from the German Research Foundation, Bonn, Germany to Herbert Fluhr (Grant FL 667/2-1).

Conflict of interest

JS declares that she has no conflict of interest. HP declares that she has no conflict of interest. FS declares that he has no conflict of interest. JR declares that he has no conflict of interest. MZ declares that he has no conflict of interest. HF declares that he has no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. This article does not contain any studies with animals performed by any of the authors.

Informed consent

Informed consent was obtained from all individual participants included in the study.

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Spratte, J., Princk, H., Schütz, F. et al. Stimulation of chemokines in human endometrial stromal cells by tumor necrosis factor-α and interferon-γ is similar under apoptotic and non-apoptotic conditions. Arch Gynecol Obstet 297, 505–512 (2018). https://doi.org/10.1007/s00404-017-4586-3

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  • DOI: https://doi.org/10.1007/s00404-017-4586-3

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