AJP - Regu Information on EB 2010
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Am J Physiol Regul Integr Comp Physiol 292: R1465-R1471, 2007. First published December 28, 2006; doi:10.1152/ajpregu.00778.2006
0363-6119/07 $8.00
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
292/4/R1465    most recent
00778.2006v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via Web of Science (10)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Hirano, S.
Right arrow Articles by Hanafusa, T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Hirano, S.
Right arrow Articles by Hanafusa, T.

INFLAMMATION AND CYTOKINES

Physiologically high concentrations of 17beta-estradiol enhance NF-{kappa}B activity in human T cells

Suzue Hirano, Daisuke Furutama, and Toshiaki Hanafusa

First Department of Internal Medicine, Osaka Medical College, Takatsuki, Japan

Submitted 7 November 2006 ; accepted in final form 20 December 2006


   ABSTRACT
TOP
 ABSTRACT
MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 GRANTS
 REFERENCES
 
Estrogen has diverse effects on inflammation and immune responses. That pregnancy is associated with remission of some autoimmune diseases and exacerbation of others suggests that physiological fluctuation in estrogen levels could affect the immune responses in humans. However, the molecular basis for these phenomena is poorly understood. We hypothesized that fluctuations of estrogen levels modulate intracellular signaling for immune responses via estrogen receptors (ERs). In reporter assays, 17beta-estradiol (E2) at a physiologically high concentration increased the activity of NF-{kappa}B in Jurkat cells stimulated by PMA/ionomycin or TNF-{alpha}. Overexpression and RNA interference experiments suggested that the effects were mediated through ERbeta. Immunoprecipitation assay showed that both ER{alpha} and ERbeta are directly associated with NF-{kappa}B in the cell nucleus. Using chromatin immunoprecipitation assay, we confirmed that ER{alpha} and ERbeta associated with NF-{kappa}B and steroid hormone coactivators at the promoter region of NF-{kappa}B regulated gene. Considering that NF-{kappa}B regulates the expression of various genes essential for cell growth and death, estrogen could regulate the fate of T cells by affecting the activity of NF-{kappa}B. To determine whether E2 alters the fate of T cells, we investigated E2 actions on T cell apoptosis, a well-known NF-{kappa}B-mediated phenomenon. E2 increased apoptosis of Jurkat cells and decreased that of human peripheral blood T cells. Our results indicate that E2 at a physiologically high concentration modulates NF-{kappa}B signaling in human T cells via ERbeta and affects T cell survival, suggesting that these actions may underlie the gender differences in autoimmune diseases.

sex hormone; autoimmune disease; coactivator; transcription factor

MANY AUTOIMMUNE DISEASES ARE more prevalent in women than in men (42, 44). Some autoimmune diseases, such as systemic lupus erythematosus (SLE), frequently show exacerbation during pregnancy (42), and in this regard, patients with SLE often have high serum estrogen concentrations (27). These clinical facts led us to hypothesize that fluctuation in estrogen levels could modulate autoimmune responses.

Baseline estrogen concentrations do not differ significantly between men and women. However, the serum concentration of 17beta-estradiol (E2), the most potent and predominant estrogen in human serum, increases from baseline 10 pM to 40 nM during pregnancy (45). Estrogen has diverse effects on inflammatory and immune systems (8, 33), and the physiological fluctuation in estrogen level could affect the immune responses in humans (5). However, the molecular basis for these phenomena is poorly understood.

NF-{kappa}B is a key regulator of inflammatory and immune systems (22). Recent studies have revealed the role of NF-{kappa}B signaling in T cells (41). NF-{kappa}B regulates the expression of various genes that control cell cycle and cell viability (16). On the other hand, estrogen elicits various biological effects in cells by binding to two distinct estrogen receptor (ER) isoforms, ER{alpha} and beta (7), members of the nuclear receptor family of ligand-dependent transcription factors. The effects of estrogen on NF-{kappa}B signaling have been mainly studied in breast cancer cells (11, 34) and studies that examined the effects of pharmacological concentrations of estrogen reported that estrogen modulates the NF-{kappa}B signaling that interacts with ERs (13, 15, 29).

The present study was designed to determine the effects of physiological fluctuations in estrogen on NF-{kappa}B signaling and the fate of immune cells. These effects may help explain the gender differences in the immune system.


   MATERIALS AND METHODS
TOP
 ABSTRACT
 MATERIALS AND METHODS
RESULTS
 DISCUSSION
 GRANTS
 REFERENCES
 
Cell culture. Human T cell line Jurkat T cells (E6-1) were purchased from Dainippon Sumitomo Pharma (Tokyo, Japan). Peripheral blood CD3+ T lymphocytes (PBT cells) from healthy donors' blood were obtained as follows. Peripheral blood mononuclear cells (PBMC) were isolated by Ficoll centrifugation and washed twice, and CD3+ cells were enriched by depletion of non-T cells using the IMag T lymphocyte enrichment set (BD Biosciences, San Jose, CA). Jurkat T Cells and PBT cells were cultured in phenol red free RPMI 1640 medium (Invitrogen, Carlsbad, CA) supplemented with 10% charcoal-stripped fetal bovine serum at 37°C in a 5% CO2 incubator. For NF-{kappa}B activation, Jurkat cells were aliquoted into a 96-well plate 40 h after transfection, cultured in a final volume of 100 µl of RPMI 1640 growth medium, and then stimulated at 37°C in a growth medium containing 60 ng/ml PMA (Sigma, St. Louis, MO) and 1 µg/ml ionomycin (Sigma) or 10 ng/ml human TNF-{alpha} (Wako, Osaka, Japan) and with the indicated concentrations of E2 (water-soluble form; Sigma).

Plasmids. The reporter plasmids, pNF-{kappa}B-Luc (Stratagene, La Jolla, CA) and pRL-TK (Promega, Madison, WI) were purchased. pOR-3, the ER{alpha} expression vector (18), was kindly supplied by Dr. P. Chambon (Institute of Genetics and Molecular and Cellular Biology, Strasbourg, France) and pSG5-ERbeta, the ERbeta expression vector (31) by Dr. J. A. Gustafsson (Department of Biosciences, Karolinska Institute, Huddinge, Sweden). Human p65 (NF-{kappa}B) cDNA was amplified by a conventional RT-PCR method and cloned into pcDNA3.1 (-)-myc-His expression vector (Invitrogen). ER{alpha} and ERbeta siRNA expression vectors for RNA interference were purchased from Panomics (Redwood City, CA).

Transient transfection and reporter assay. Jurkat cells were cotransfected with pNF{kappa}B-Luc and the indicated expression plasmids using the method as explained by the manufacturer (Amaxa Biosystems, Cologne, Germany). pRL-TK, which expresses Renilla luciferase, was also cotransfected as an internal control. After 6-h stimulation with PMA/ionomycin or TNF-{alpha}, cells were lysed and assayed using Dual-Glo Luciferase Assay System (Promega). Luciferase activity was read on the luminometer (Luminoskan Ascent; Thermo Electron, Waltham, MA) and determined at least three times for each experimental condition. The fold induction of luciferase activity (firefly luciferase/Renilla luciferase) was expressed relative to the baseline activity in untreated cells.

ELISA. Jurkat cells and PBT cells were treated with 10 ng/ml TNF-{alpha} and the indicated concentrations of E2 for 6 h. Cells were washed twice and lysed with radioimmunoprecipitation (RIPA) buffer (1% Nonidet P-40, 0.5% sodium deoxycholate, 0.1% SDS, pH 7.4, in PBS). The level of activated NF-{kappa}B was quantified in cell lysate by using the NF-{kappa}B-p65 ELISA Kit (Nventa Biopharmaceuticals, San Diego, CA).

Immunoprecipitation and Western blot analysis. For immunoprecipitation assay, p65 and ER{alpha} or ERbeta were overexpressed in cells. Forty hours after transfection, cells were treated for 6 h with 10 nM E2 and 10 ng/ml TNF-{alpha}. Nuclear proteins were obtained using Nuclear Complex Co-IP Kit (Active Motif, Carlsbad, CA). Using agarose-conjugated anti-p65 (Santa Cruz Biotechnology, Santa Cruz, CA), anti-ER{alpha} (Lab Vision, Fremont, CA), anti-ERbeta (Santa Cruz) or control rabbit IgG antibodies, we examined the status of coprecipitation of ER{alpha} or ERbeta with p65. Proteins were separated by SDS-PAGE on 10% gels and transferred onto a polyvinylidene difluoride membrane (Millipore, Bedford, MA). The membrane was blocked overnight with 5% skim milk, followed by incubation with anti-ER{alpha} (Lab Vision) or anti-ERbeta antibody (Lab Vision) for 2 h at room temperature. After washing, the horseradish peroxidase-labeled anti-mouse secondary antibody (Chemicon International, Temecula, CA) was added for 2 h. The proteins were visualized with the ECL substrate kit (Amersham Life Science, Buckinghamshire, UK).

Chromatin Immunoprecipitation assays. Jurkat cells were stimulated with 10 nM E2 and 10 ng/ml TNF-{alpha} for 6 h. DNA was purified with chromatin immunoprecipitation (ChIP)-IT Kit (Active Motif). Protein/DNA complexes were immunoprecipitated with control mouse IgG (included in the ChIP-IT Kit), anti-ER{alpha} antibody (Lab Vision), anti-ERbeta antibody, anti-p65 antibody, anti-SRC1 antibody, and anti-GRIP1 antibody (Santa Cruz). The primers used for the IL-4 promoter were (-336)-5'-CAACAAATTCGGACACCTG-3' and (-40)-5'-GCTGAAACCGAGGGAAAAT-3' (a 296-bp product) (23).

Apoptosis assay. Jurkat cells and PBT cells were pretreated with or without 10 nM E2 or 10 mM ICI 182780 (Tocris Cookson, Ellisville, MO) for 6 h. Jurkat cells were then plated in anti-human CD3 plate (BD Biosciences) for 36 h to induce the activation-induced cell death (AICD) (6, 14, 25). On the other hand, PBT cells were cultured for 36 h in a medium containing 4 µg/ml Concanavalin A (Sigma) (43). Cells were analyzed with Annexin V Fluos kit (Roche, Basel, Switzerland) by a flow cytometer (FACS Aria; BD Biosciences). Cells were stained with Annexin V-Fluos and propidium iodide (PI). Apoptotic cells were defined as Annexin V-positive and PI-negative cells.

Statistical analysis. Data are expressed as means ± SD of at least three independent experiments with triplicate estimations per data point. P values were calculated by ANOVA.


   RESULTS
TOP
 ABSTRACT
 MATERIALS AND METHODS
 RESULTS
DISCUSSION
 GRANTS
 REFERENCES
 
E2 enhances NF-{kappa}B reporter activity in Jurkat cells. To investigate the actions of estrogen on intracellular signaling, we screened changes in activities of various transcription factors with reporter plasmids in Jurkat cells (data not shown), and we then focused on NF-{kappa}B signaling. First, we examined the NF-{kappa}B activity in Jurkat cells with reporter plasmid. The results showed E2 alone did not affect the NF-{kappa}B signaling in these cells (Fig. 1). However, when the cells were stimulated in the presence of phorbol ester PMA and ionomycin, known as strong activators of NF-{kappa}B in T cells (24, 38), physiologically high concentrations (10 nM) of E2, representing the levels in pregnancy, enhanced the activated NF-{kappa}B signaling (Fig. 2A). After stimulation by TNF-{alpha}, another activator of NF-{kappa}B in T cells, E2 also showed a similar effect on Jurkat cells (Fig. 2B).


Figure 1
View larger version (8K):
[in this window]
[in a new window]

  		Fig. 1. 17beta-estradiol (E2) alone does not change NF-{kappa}B signaling. pNF-{kappa}B-Luc was transfected into Jurkat cells. Jurkat cells were treated with the indicated concentrations of E2 for 6 h, and luciferase activity was measured. Data are expressed as means ± SD.

 

Figure 2
View larger version (10K):
[in this window]
[in a new window]

  		Fig. 2. E2 enhances the NF-{kappa}B reporter activity in Jurkat cells by two different stimuli. pNF-{kappa}B-Luc was transfected into Jurkat cells and the cells were treated with the indicated concentrations of E2 and activated by 60 ng/ml PMA and 1 µg/ml ionomycin (A), or 10 ng/ml TNF-{alpha} (B) for 6 h. *P < 0.05, **P < 0.01.

 
E2 enhances activation of NF-{kappa}B in human T cells. The major form of NF-{kappa}B is known as a heterodimer of p65 and p50 (22). As indicated above, E2 enhanced activation of NF-{kappa}B signaling in Jurkat cells with the reporter assay system. To confirm the results, we next quantified the activated p65 in Jurkat and PBT cells with ELISA. The assay uses NF-{kappa}B consensus sequence to capture only the active forms of NF-{kappa}B, because only active forms of NF-{kappa}B can translocate from the cytoplasm to nucleus and bind the genomic DNA. In both Jurkat and PBT cells, TNF-{alpha} induced active forms of p65 were increased in the presence of physiologically high concentration of E2 (10 nM) but not with physiologically low concentration (10 pM) or absence of E2 (Fig. 3).


Figure 3
View larger version (9K):
[in this window]
[in a new window]

  		Fig. 3. E2 enhances activation of NF-{kappa}B in human T cells. Activated p65 was measured by ELISA in Jurkat (A) and human peripheral blood T cells (PBT) (B). Cells were treated with the indicated concentrations of E2 and stimulated with 10 ng/ml TNF-{alpha} for 6 h. *P < 0.01, **P < 0.001.

 
E2 enhances activation of NF-{kappa}B in Jurkat cells via ERbeta. Several studies have reported ER expression in immune cells (32, 37), and a recent study suggested that ERbeta is predominantly expressed in human T cells (37). Our results also showed the mRNA expression of both ER{alpha} and ERbeta in both Jurkat cells and human PBT cells and the predominance of ERbeta (data not shown). Since the effects of estrogen are mediated mainly by binding to ERs, we transiently transfected Jurkat cells with ER expression vectors or ER siRNA vectors and determined the ER subtype that could affect the enhancement of activation of NF-{kappa}B signaling in human T cells. In these experiments, Jurkat cells were stimulated with PMA and ionomycin followed by analysis of luciferase activity. As shown in Figs. 2 and 3, 10 nM E2 enhanced activated NF-{kappa}B signaling with endogenous ERs. Under physiologically high E2 condition, overexpression of ERbeta enhanced NF-{kappa}B activation (Fig. 4B), while overexpression of ER{alpha} inhibited NF-{kappa}B activation (Fig. 4A). On the other hand, the results of RNA interference experiments for endogenous ERs (Fig. 4, C and D) showed that physiologically high E2 did not enhance NF-{kappa}B activation in ERbeta-knockdown cells. These results suggest that the effect of E2 on NF-{kappa}B signaling is mediated mainly through ERbeta and not ER{alpha}.


Figure 4
View larger version (15K):
[in this window]
[in a new window]

  		Fig. 4. E2 enhances activation of NF-{kappa}B via ERbeta. Jurkat cells were cotransfected with pN{kappa}B-Luc and with estrogen receptor (ER){alpha} (A) or ERbeta (B) expression vector, and with ER{alpha} (C) or ERbeta (D) siRNA vector for RNA interference. Cells were treated with 60 ng/ml PMA, 1 µg/ml ionomycin and the indicated concentrations of E2 for 6 h. *P < 0.05 compared with the basal condition.

 
NF-{kappa}B interacts with ERs and steroid hormone coactivators on the promoter region of NF-{kappa}B target gene in T cell nucleus. To investigate the mechanisms of estrogen effects on NF-{kappa}B signaling, we examined the direct interaction between ER and NF-{kappa}B under stimulation with TNF-{alpha} and E2. After cotransfection of p65 expression vector and ER{alpha} or ER-beta expression vector, nuclear extracts of cells were immunoprecipitated with p65 antibody or control antibodies and the proteins were immunoblotted with anti-ER{alpha} or ERbeta antibody (Fig. 5, A and B). Activated p65 of NF-{kappa}B interacted with both ER{alpha} and ERbeta in cell nucleus. To further investigate the association between ERs and NF-{kappa}B, we conducted ChIP assays in Jurkat cells. ChIP assays are used to identify the in vivo position of transcription factors on cognate DNA sites (19). We examined the binding of NF-{kappa}B/ER complex on IL-4 promoter, a NF-{kappa}B regulated gene (23), under TNF-{alpha}- and E2-stimulated conditions. Both endogenous ER{alpha} and ERbeta bound to the IL-4 promoter in vivo (Fig. 5C, lanes 3 and 4). We also examined steroid hormone coactivators, steroid receptor coactivator1 (SRC1/NCOA1) and glucocorticoid receptor interacting protein1 (GRIP1/NCOA2) (9, 39). Since both coactivators are known to interact with nuclear receptors (26), we confirmed that there are no steroid hormone response elements (including estrogen response elements) on the IL-4 promoter region with MATCH software public version 1.0 (46). SRC1 and GRIP1 were precipitated concurrently with NF-{kappa}B and ERs on IL-4 promoter (Fig. 5C, lanes 5 and 6), suggesting that they form a large transcriptional complex on the promoter region.


Figure 5
View larger version (24K):
[in this window]
[in a new window]

  		Fig. 5. NF-{kappa}B (p65) interacts with ERs and steroid hormone coactivators on a NF-{kappa}B regulated gene promoter in activated Jurkat cells. C2C12 cells were cotransfected with p65 and ER{alpha} expression vectors (A), while Jurkat cells were cotransfected with p65 and ERbeta expression vectors (B). Cells were treated with 10 nM E2 and stimulated with 10 ng/ml TNF-{alpha} for 6 h. Nuclear extracts were immunoprecipitated with antibodies and immunoblotted with anti-ER{alpha} (A) or anti-ERbeta antibody (B). C: Jurkat cells stimulated with 10 nM E2 and 10 ng/ml TNF-{alpha} for 6 h were subjected to chromatin immunoprecipitation analysis. DNA/protein complexes were immunoprecipitated with control antibody (normal mouse IgG) (lane 1), anti-p65 antibody (lane 2), anti-ER{alpha} antibody (lane 3), anti-ERbeta antibody (lane 4), anti-GRIP1 antibody (lane 5), or anti-SRC1 antibody (lane 6). The specific 296 bp product of the IL-4 promoter was amplified by PCR with the immunoprecipitated DNA/protein complexes and (lane 7) input DNA.

 
E2 modulates apoptosis of T cells. To examine whether estrogen affects the T cell fate through NF-{kappa}B, we analyzed AICD in Jurkat cells, which is mediated by NF-{kappa}B (25). E2 is known to induce apoptosis in Jurkat cells (12). Treatment with a physiologically high concentration of E2 (10 nM) increased the proportion of apoptotic Jurkat cells (Fig. 6A). On the other hand, treatment of PBT cells with the same concentration of E2 rescued these cells from apoptosis (Fig. 6B), in agreement with a previous study that showed inhibition of NF-{kappa}B activity resulted in increased PBT cell apoptosis through concanavalin A stimulation (43). To investigate whether these effects are mediated via ERs, we next examined effects of ICI-182780, an ER antagonist (1). Inhibition of endogenous ER action with ICI-182780 decreased apoptosis by 10% in Jurkat and increased by 31% in PBT cells. These results indicate that the estrogen receptor signaling affects the NF-{kappa}B signaling in human T cells and consequently affects the T cell fate.


Figure 6
View larger version (15K):
[in this window]
[in a new window]

  		Fig. 6. Physiologically high concentration of E2 modulates NF-{kappa}B-mediated apoptosis/survival of human T cells. A: Jurkat cells were pretreated with or without 10 nM E2 for 6 h and plated onto anti-human CD3 plate for 36 h to induce activation-induced cell death. B: PBT cells were pretreated with or without 10 nM E2 for 6 h and stimulated with 4 µg/ml Concanavalin A for 36 h. Cells were analyzed by flow cytometry. Numbers in figures represent %apoptotic cells. Apoptotic cells were defined as Annexin V-positive and propidium iodide-negative cells. These data are representatives of 3 independent experiments.

 

   DISCUSSION
TOP
 ABSTRACT
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
GRANTS
 REFERENCES
 
Our study demonstrated that E2 enhances NF-{kappa}B activity in human T cells. E2 modulated NF-{kappa}B signaling in both Jurkat cells and human PBT cells when stimulated. In T cells, E2 seems to act through ERbeta, and the direct interaction among ERs, NF-{kappa}B, and steroid hormone coactivators could modulate the NF-{kappa}B signaling.

To our knowledge, this is the first report on the functional relationship between estrogen and NF-{kappa}B in human T cells and on the association among ERs, NF-{kappa}B, and steroid hormone coactivators. Our results suggest that the effect of E2 is mediated via ERbeta in T cells. Several studies have examined estrogen actions on NF-{kappa}B activity, mainly in breast cancer cells (11, 34, 36). However, the results of these studies are controversial. Liganded ER{alpha} is reported to inhibit NF-{kappa}B activity in an estrogen-dependent manner at nanomolar concentrations of estrogen in various cell lines, such as U937 and MCF-7 cells, and ERbeta is also known to have an inhibitory effect on NF-{kappa}B activity (15). On the other hand, no antagonism between ER and NF-{kappa}B was reported in a study of other breast cancer cells, T47D and HC11 cells treated with TNF-{alpha} and E2 (36). Furthermore, another study showed that E2 enhanced NF-{kappa}B transactivation via ERbeta in human embryonic kidney HEK-293 cells and prostate cancer DU145 cells (21). These results suggest that estrogen actions on NF-{kappa}B activity may depend on ER and cell subtypes. Although there are also many controversial results about expression of ER genes in immune cells (32, 37), the latest report indicates that ERbeta is the predominant ER in human peripheral blood leucocytes (37). Our results are consistent with that study, i.e., estrogen could affect the NF-{kappa}B activity in human T cells via ERbeta.

Our results showed E2 alone did not change NF-{kappa}B signaling in Jurkat cells and human PBT cells, however; it enhanced the NF-{kappa}B signaling when activated. Though the effects between two different concentrations of E2 were modest, our report focused on the physiological fluctuations and that slight changes in physiological circumstances could affect or trigger subsequent events.

In addition, we obtained similar results in experiments of both PMA/ionomycin-activated and TNF-{alpha}-activated NF-{kappa}B signaling. In the NF-{kappa}B activation pathway, it was reported that PMA and ionomycin act on protein kinase C (30), and TNF-{alpha} acts on caspase-8 (28). Therefore, we suggest that E2 modulates the NF-{kappa}B signaling downstream of these points, i.e., at the nucleus. Using immunoprecipitation assay, we demonstrated a direct interaction between ERs and p65 in cell nucleus. With ChIP assays, liganded ER{alpha} and ERbeta associated with NF-{kappa}B on the NF-{kappa}B regulated gene promoter. Moreover, steroid coactivators SRC1 and GRIP1 were confirmed to associate with NF-{kappa}B under physiologically high E2 conditions. These results suggest that the E2-liganded ERs associate with steroid coactivators and simultaneously interact with NF-{kappa}B on the NF-{kappa}B regulated gene promoters, and that recruitment of steroid hormone coactivators could affect the transcriptional activity of NF-{kappa}B (Fig. 7).


Figure 7
View larger version (42K):
[in this window]
[in a new window]

  		Fig. 7. Model of E2-enhanced NF-{kappa}B activity. After E2 binding to ER, ER translocates to nucleus and binds to p65. Concurrently, ER recruits steroid hormone coactivators to the ER-NF-{kappa}B complex on the NF-{kappa}B target gene promoter region, resulting in upregulation of the transactivity.

 
Physiologically high concentrations of E2, equivalent to those in pregnancy, altered apoptosis of human T cells. Using Jurkat T cells and PBT cells, we showed here that E2 increased apoptosis of Jurkat cells but rescued PBT cells from apoptosis. NF-{kappa}B has been reported as a contextual regulator of apoptosis (24). It was reported also that NF-{kappa}B contributed to AICD by upregulation of Fas ligand expression in Jurkat T cells (17), whereas NF-{kappa}B in PBT cells is required for T cell survival (43). We believe that distinct signaling pathways mediate T cell apoptosis under different contexts in these two types of T cells.

The significance of estrogen in human T cells is still not clear. There are several reports on the contribution of the Rel/NF-{kappa}B family on the development of Th1 and Th2 lymphocytes (4, 10). Pregnancy is associated with exacerbation of SLE, a Th2 disease, but with remission of multiple sclerosis (MS) and rheumatoid arthritis, known as Th1 diseases (20, 44). Previous reports suggested that E2 reduces pathological Th1 responses (20, 33). Moreover, several studies reported that E2 administration accelerated disease activities in murine models of SLE (3, 35, 40), but inhibited experimental autoimmune encephalomyelitis, an animal model of MS (2). Autoreactive cells that escape immunosurveillance can result in many autoimmune diseases. In the present study, we demonstrated that E2 enhanced activation of NF-{kappa}B, a key regulator of cytokine production. Although further studies are needed to address the underlying mechanisms of gender differences in the immune system, these results could help explain inappropriate homeostasis of the immune system.


   GRANTS
TOP
 ABSTRACT
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 GRANTS
REFERENCES
 
This work was supported by a grant from the Osaka Medical Research Foundation for Incurable Diseases and the Japan Society for the Promotion of Science (to D. Furutama).


   ACKNOWLEDGMENTS
 
The authors thank Masa-aki Shibata from the Department of Anatomy and Cell Biology, Osaka Medical College, for his help on Jurkat transfection. We acknowledge the helpful advice of Shigeki Makino.


   FOOTNOTES
 

Address for reprint requests and other correspondence: D. Furutama, First Dept. of Internal Medicine, Osaka Medical College, 2-7 Daigaku-machi, Takatsuki 569-8686, Japan (e-mail: in1003{at}poh.osaka-med.ac.jp)

The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.


   REFERENCES
TOP
 ABSTRACT
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 GRANTS
 REFERENCES
 

  1. Ameller T, Legrand P, Marsaud V, Renoir JM. Drug delivery systems for oestrogenic hormones and antagonists: the need for selective targeting in estradiol-dependent cancers. J Steroid Biochem Mol Biol 92: 1–18, 2004.[CrossRef][Web of Science][Medline]
  2. Bebo BF Jr, Fyfe-Johnson A, Adlard K, Beam AG, Vandenbark AA, Offner H. Low-dose estrogen therapy ameliorates experimental autoimmune encephalomyelitis in two different inbred mouse strains. J Immunol 166: 2080–2089, 2001.[Abstract/Free Full Text]
  3. Carlsten H, Tarkowski A, Holmdahl R, Nilsson LA. Oestrogen is a potent disease accelerator in SLE-prone MRL lpr/lpr mice. Clin Exp Immunol 80: 467–473, 1990.[Web of Science][Medline]
  4. Corn RA, Aronica MA, Zhang F, Tong Y, Stanley SA, Kim SR, Stephenson L, Enerson B, McCarthy S, Mora A, Boothby M. T cell-intrinsic requirement for NF-{kappa}B induction in postdifferentiation IFN-{gamma} production and clonal expansion in a Th1 response. J Immunol 171: 1816–1824, 2003.[Abstract/Free Full Text]
  5. Cutolo M, Sulli A, Capellino S, Villaggio B, Montagna P, Seriolo B, Straub RH. Sex hormones influence on the immune system: basic and clinical aspects in autoimmunity. Lupus 13: 635–638, 2004.[Abstract/Free Full Text]
  6. Dhein J, Walczak H, Baumler C, Debatin KM, Krammer PH. Autocrine T-cell suicide mediated by APO-1/(Fas/CD95). Nature 373: 438–441, 1995.[CrossRef][Medline]
  7. Enmark E, Gustafsson JA. Oestrogen receptors–an overview. J Intern Med 246: 133–138, 1999.[CrossRef][Web of Science][Medline]
  8. Grimaldi CM, Cleary J, Dagtas AS, Moussai D, Diamond B. Estrogen alters thresholds for B cell apoptosis and activation. J Clin Invest 109: 1625–1633, 2002.[CrossRef][Web of Science][Medline]
  9. Halachmi S, Marden E, Martin G, MacKay H, Abbondanza C, Brown M. Estrogen receptor-associated proteins: possible mediators of hormone-induced transcription. Science 264: 1455–1458, 1994.[Abstract/Free Full Text]
  10. Hilliard BA, Mason N, Xu L, Sun J, Lamhamedi-Cherradi SE, Liou HC, Hunter C, Chen YH. Critical roles of c-Rel in autoimmune inflammation and helper T cell differentiation. J Clin Invest 110: 843–850, 2002.[CrossRef][Web of Science][Medline]
  11. Hsu SM, Chen YC, Jiang MC. 17 beta-estradiol inhibits tumor necrosis factor-{alpha}-induced nuclear factor-{kappa}B activation by increasing nuclear factor-{kappa}B p105 level in MCF-7 breast cancer cells. Biochem Biophys Res Commun 279: 47–52, 2000.[CrossRef][Web of Science][Medline]
  12. Jenkins JK, Suwannaroj S, Elbourne KB, Ndebele K, McMurray RW. 17beta-estradiol alters Jurkat lymphocyte cell cycling and induces apoptosis through suppression of Bcl-2 and cyclin A. Int Immunol 1: 1897–1911, 2001.
  13. Jordan VC. Selective estrogen receptor modulation: concept and consequences in cancer. Cancer Cell 5: 207–213, 2004.[CrossRef][Web of Science][Medline]
  14. Ju ST, Panka DJ, Cui H, Ettinger R, el-Khatib M, Sherr DH, Stanger BZ, Marshak-Rothstein A. Fas(CD95)/FasL interactions required for programmed cell death after T-cell activation. Nature 373: 444–448, 1995.[CrossRef][Medline]
  15. Kalaitzidis D, Gilmore TD. Transcription factor cross-talk: the estrogen receptor and NF-kappaB. Trends Endocrinol Metab 16: 46–52, 2005.[CrossRef][Web of Science][Medline]
  16. Karin M, Lin A. NF-{kappa}B at the crossroads of life and death. Nat Immun 3: 221–227, 2002.
  17. Kasibhatla S, Genestier L, Green DR. Regulation of fas-ligand expression during activation-induced cell death in T lymphocytes via nuclear factor {kappa}B. J Biol Chem 274: 987–992, 1999.[Abstract/Free Full Text]
  18. Kumar V, Green S, Stack G, Berry M, Jin JR, Chambon P. Functional domains of the human estrogen receptor. Cell 51: 941–951, 1987.[CrossRef][Web of Science][Medline]
  19. Kuo MH, Allis CD. In vivo cross-linking and immunoprecipitation for studying dynamic Protein:DNA associations in a chromatin environment. Methods 19: 425–433, 1999.[CrossRef][Web of Science][Medline]
  20. Lang TJ. Estrogen as an immunomodulator. Clin Immunol 113: 224–230, 2004.[CrossRef][Web of Science][Medline]
  21. Leung YK, Gao Y, Lau KM, Zhang X, Ho SM. ICI 182,780-regulated gene expression in DU145 prostate cancer cells is mediated by estrogen receptor-beta/NF-{kappa}B crosstalk. Neoplasia 8: 242–249, 2006.[CrossRef][Web of Science][Medline]
  22. Li Q, Verma IM. NF-{kappa}B regulation in the immune system. Nat Rev Immunol 2: 725–734, 2002.[CrossRef][Web of Science][Medline]
  23. Li-Weber M, Giaisi M, Baumann S, Palfi K, Krammer PH. NF-{kappa}B synergizes with NF-AT and NF-IL6 in activation of the IL-4 gene in T cells. Eur J Immunol 34: 1111–1118, 2004.[CrossRef][Web of Science][Medline]
  24. Lin B, Williams-Skipp C, Tao Y, Schleicher MS, Cano LL, Duke RC, Scheinman RI. NF-{kappa}B functions as both a proapoptotic and antiapoptotic regulatory factor within a single cell type. Cell Death Differ 6: 570–582, 1999.[CrossRef][Web of Science][Medline]
  25. Maher SG, Condron CE, Bouchier-Hayes DJ, Toomey DM. Taurine attenuates CD3/interleukin-2-induced T cell apoptosis in an in vitro model of activation-induced cell death (AICD). Clin Exp Immunol 139: 279–286, 2005.[CrossRef][Web of Science][Medline]
  26. McKenna NJ, O'Malley BW. Combinatorial control of gene expression by nuclear receptors and coregulators. Cell 108: 465–474, 2002.[CrossRef][Web of Science][Medline]
  27. McMurray RW, May W. Sex hormones and systemic lupus erythematosus: review and meta-analysis. Arthritis Rheum 48: 2100–2110, 2003.[CrossRef][Web of Science][Medline]
  28. Nagata S. Apoptosis by death factor. Cell 88: 355–365, 1997.[CrossRef][Web of Science][Medline]
  29. Nakshatri H, Bhat-Nakshatri P, Martin DA, Goulet RJ Jr, Sledge GW Jr. Constitutive activation of NF-{kappa}B during progression of breast cancer to hormone-independent growth. Mol Cell Biol 17: 3629–3639, 1997.[Abstract]
  30. Nishizuka Y. The role of protein kinase C in cell surface signal transduction and tumour promotion. Nature 308: 693–698, 1984.[CrossRef][Medline]
  31. Ogawa S, Inoue S, Watanabe T, Hiroi H, Orimo A, Hosoi T, Ouchi Y, Muramatsu M. The complete primary structure of human estrogen receptor beta (hER beta) and its heterodimerization with ER alpha in vivo and in vitro. Biochem Biophys Res Commun 243: 122–126, 1998.[CrossRef][Web of Science][Medline]
  32. Phiel KL, Henderson RA, Adelman SJ, Elloso MM. Differential estrogen receptor gene expression in human peripheral blood mononuclear cell populations. Immunol Lett 97: 107–113, 2005.[CrossRef][Web of Science][Medline]
  33. Polanczyk MJ, Carson BD, Subramanian S, Afentoulis M, Vandenbark AA, Ziegler SF, Offner H. Cutting edge: estrogen drives expansion of the CD4+CD25+ regulatory T cell compartment. J Immunol 173: 2227–2230, 2004.[Abstract/Free Full Text]
  34. Pratt MA, Bishop TE, White D, Yasvinski G, Menard M, Niu MY, Clarke R. Estrogen withdrawal-induced NF-kappaB activity and bcl-3 expression in breast cancer cells: roles in growth and hormone independence. Mol Cell Biol 23: 6887–6900, 2003.[Abstract/Free Full Text]
  35. Roubinian JR, Talal N, Greenspan JS, Goodman JR, Siiteri PK. Effect of castration and sex hormone treatment on survival, anti-nucleic acid antibodies, and glomerulonephritis in NZB/NZW F1 mice. J Exp Med 147: 1568–1583, 1978.[Abstract/Free Full Text]
  36. Rubio MF, Werbajh S, Cafferata EG, Quaglino A, Colo GP, Nojek IM, Kordon EC, Nahmod VE, Costas MA. TNF-{alpha} enhances estrogen-induced cell proliferation of estrogen-dependent breast tumor cells through a complex containing nuclear factor-{kappa}B. Oncogene 25: 1367–1377, 2006.[CrossRef][Web of Science][Medline]
  37. Shim GJ, Gherman D, Kim HJ, Omoto Y, Iwase H, Bouton D, Kis LL, Andersson CT, Warner M, Gustafsson JA. Differential expression of oestrogen receptors in human secondary lymphoid tissues. J Pathol 208: 408–414, 2006.[CrossRef][Web of Science][Medline]
  38. Siebenlist U, Brown K, Claudio E. Control of lymphocyte development by nuclear factor-{kappa}B. Nat Rev Immunol 5: 435–445, 2005.[CrossRef][Web of Science][Medline]
  39. Smith CL, O'Malley BW. Coregulator function: a key to understanding tissue specificity of selective receptor modulators. Endocr Rev 25: 45–71, 2004.[Abstract/Free Full Text]
  40. Steinberg AD, Melez KA, Raveche ES, Reeves JP, Boegel WA, Smathers PA, Taurog JD, Weinlein L, Duvic M. Approach to the study of the role of sex hormones in autoimmunity. Arthritis Rheum 22: 1170–1176, 1979.[Web of Science][Medline]
  41. van Oers NS, Chen ZJ. Cell biology. Kinasing and clipping down the NF-{kappa}B trail. Science 308: 65–66, 2005.[Abstract/Free Full Text]
  42. Wallace DJ, Hahn BH. Dubois' Lupus Erythematosus. Baltimore, MD: Williams & Wilkins, 1997.
  43. Wan YY, DeGregori J. The survival of antigen-stimulated T cells requires NFkappaB-mediated inhibition of p73 expression. Immunity 18: 331–342, 2003.[CrossRef][Web of Science][Medline]
  44. Whitacre CC, Reingold SC, O'Looney PA. A gender gap in autoimmunity. Science 283: 1277–1278, 1999.[Free Full Text]
  45. Williams RH, Larsen PR, Kronenberg HM, Melmed S, Polonsky KS. Williams Textbook of Endocrinology. Philadelphia, PA: Saunders, 2002.
  46. Wingender E, Chen X, Fricke E, Geffers R, Hehl R, Liebich I, Krull M, Matys V, Michael H, Ohnhauser R, Pruss M, Schacherer F, Thiele S, Urbach S. The TRANSFAC system on gene expression regulation. Nucleic Acids Res 29: 281–283, 2001.[Abstract/Free Full Text]




This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
292/4/R1465    most recent
00778.2006v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via Web of Science (10)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Hirano, S.
Right arrow Articles by Hanafusa, T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Hirano, S.
Right arrow Articles by Hanafusa, T.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Visit Other APS Journals Online
Copyright © 2007 by the American Physiological Society.