17Beta-estradiol suppresses TLR3-induced cytokine and chemokine production in endometrial epithelial cells.
BACKGROUND
The human endometrium is an important site for contact between the host and pathogens ascending the reproductive tract, and thus plays an important role in female reproductive tract immunity. Previous work in our laboratory has suggested that Toll-like receptors (TLRs) are involved in endometrial epithelial recognition of pathogens and that ligation of endometrial TLRs results in the production of cytokines and chemokines important for both immune and reproductive functions of the endometrium. We have also demonstrated cyclic regulation of TLR3 mRNA and protein expression in human endometrium, suggesting that steroid hormones might play a role in the expression and function of TLR3. In this study, the effects of 17beta-estradiol (E2) and progesterone (P) on TLR3 expression and function in endometrial cell lines were investigated.


METHODS
Endometrial epithelial cell lines were cultured and examined for the presence of TLR3 and hormone receptors by endpoint RT-PCR. For hormonal studies, cells were pre-treated with ethanol vehicle, 10(-8) M E2, and/or 10(-7) M P. For antagonist assays, cells were treated with the ER antagonist, ICI 182, 780, or the PR antagonist, RU486, for two hours prior to treatment with hormones. Following hormone or hormone/antagonist pre-treatment, cells were stimulated with vehicle, the synthetic TLR3 ligand, polyinosinic-polycytidylic acid (Poly I:C), a negative dsDNA control, or a positive control. Cytokine and chemokine production post-stimulation was measured by ELISA. The effects of E2 and P on TLR3 mRNA and protein expression were measured using Real Time RT-PCR and FACS analysis, respectively.


RESULTS
Stimulation of TLR3-expressing cells with the synthetic TLR3 ligand, Poly I:C, resulted in the production of cytokines and chemokines important for endometrial function and regulation. Suppression of Poly I:C-induced cytokine and chemokine production by cells treated with 10(-8) M E2, but not cells treated with 10(-7) M P, was observed in endometrial epithelial cell lines expressing TLR3 and estrogen receptor alpha (ERalpha). The effects of E2 were not observed on cells which did not express ERalpha or in cells pre-treated with the ER antagonist, ICI 182, 780. Treatment with E2 did not affect TLR3 mRNA or protein expression. However, treatment with E2 did suppress cytokine and chemokine production resulting from TLR3 stimulation with Poly I:C, suggesting that E2 modulates TLR3 function.


CONCLUSIONS
The data presented in this study are the first indication that E2 can markedly alter the innate immune response to dsRNA, providing a previously unreported process by which E2 can alter immune responses.
Lesmeister
Margaret J
MJ
Department of Molecular Microbiology and Immunology, University of Missouri-Columbia, School of Medicine, MO, USA. mjl8ce@mizzou.edu
Jorgenson
Rebecca L
RL
Young
Steven L
SL
Misfeldt
Michael L
ML
D002460:Cell Line; D045744:Cell Line, Tumor; D018925:Chemokines / Q000096:biosynthesis*; D016207:Cytokines / Q000096:biosynthesis*; D004717:Endometrium / Q000166:cytology; D004847:Epithelial Cells; D004958:Estradiol / Q000031:analogs & derivatives / Q000494:pharmacology*; D005260:Female; D000077267:Fulvestrant; D006801:Humans; D015735:Mifepristone / Q000494:pharmacology; D011070:Poly I-C / Q000037:antagonists & inhibitors / Q000494:pharmacology; D011089:Polydeoxyribonucleotides / Q000494:pharmacology; D011374:Progesterone / Q000494:pharmacology; D011960:Receptors, Estrogen / Q000096:biosynthesis; D011980:Receptors, Progesterone / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D051196:Toll-Like Receptor 3 / Q000096:biosynthesis* / Q000187:drug effects
D018925:Chemokines; D016207:Cytokines; D011089:Polydeoxyribonucleotides; D011960:Receptors, Estrogen; D011980:Receptors, Progesterone; C495344:TLR3 protein, human; D051196:Toll-Like Receptor 3; D000077267:Fulvestrant; C031156:poly d(I-C); D015735:Mifepristone; D011374:Progesterone; D004958:Estradiol; D013755:Tetradecanoylphorbol Acetate; D011070:Poly I-C