Journal of Reproduction & Infertility

Journal of Reproduction & Infertility

Lipopolysaccharide- and Lipoteichoic Acid-mediated Pro-inflammatory Cytokine Production and Modulation of TLR2, TLR4 and MyD88 Expression in Human Endometrial Cells

Authors
Nesa Rashidi 1
Mahroo Mirahmadian 1
Mahmood Jeddi-Tehrani 2
Simin Rezania 3
Jamileh Ghasemi 4
Somaieh Kazemnejad 5
Ebrahim Mirzadegan 6
Sedigheh Vafaei 4
Maryam Kashanian 7
Zahra Rasoulzadeh 8
Amir Hassan Zarnani 9
1 Immunology Department, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran
2 Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
3 Biophysics Institute, Medical University of Graz, Graz, Austria
4 Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
5 Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
6 Reproductive Immunology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
7 Department of Obstetrics and Gynecology, Shahid Akbarabadi Hospital, Faculty of Medicine, Iran university of Medical Sciences, Tehran, Iran
8 Department of Immunology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran
9 Reproductive Immunology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran; Immunology Research Center, Iran University of Medical Sciences, Tehran, Iran
Abstract
Background: Toll-like receptor (TLR)-mediated inflammatory processes are supposed to be involved in pathophysiology of spontaneous abortion and preterm labor. Here, we investigated functional responses of human endometrial stromal cells (ESCs) and whole endometrial cells (WECs) to lipopolysaccharide (LPS) and lipoteichoic acid (LTA). Methods: Endometrial tissues were obtained from 15 cycling women who underwent laparoscopic tubal ligation. Modulation of TLR2, TLR4 and MyD88 expression and pro-duction of pro-inflammatory cytokines by WECs and ESCs in response to LPS and LTA were assessed. Results: WECs and ESCs expressed significant levels of TLR4 and MyD88 transcripts but, unlike WECs, ESCs failed to express TLR2 gene. Regardless of positive results of Western blotting, ESCs did not express TLR4 at their surface as judged by flow cytometry. Immunofluorescent staining revealed intracellular localization of TLR4 with predominant perinuclear pattern. LPS stimulation marginally increased TLR4 gene ex-pression in both cell types, whereas such treatment significantly upregulated MyD88 gene expression after 8 hr (p<0.05). At the protein level, however, LPS activation significantly increased TLR4 expression by ESCs (p<0.05). LTA stimulation of WECs was accompanied with non-significant increase of TLR2 and MyD88 transcripts. LPS and LTA stimulation of WECs caused significant production of IL-6 and IL-8 in a dose-dependent manner (p<0.05). Similarly, ESCs produced significant amounts of IL-6, IL-8 and also TNF-α in response to LPS activation (p<0.05). Conclusion: Our results provided further evidence of initiation of inflammatory processes following endometrial TLR activation by bacterial components which could potentially be harmful to developing fetus.
Keywords
cytokine
Endometrium
Inflammation
LPS
LTA
toll
like receptor
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Journal of Reproduction & Infertility
Volume 16, Issue 2
Spring 2015
Pages 72-81