Differential Regulation of the Expression of CD95 Ligand, Receptor Activator of Nuclear Factor-κB Ligand (RANKL), TNF-Related Apoptosis-Inducing Ligand (TRAIL), and This information is current as TNF-α During T Cell Activation of September 25, 2021. Ruoxiang Wang, Liying Zhang, Xiaoren Zhang, Jose Moreno, Xunyi Luo, Mehrdad Tondravi and Yufang Shi J Immunol 2001; 166:1983-1990; ; doi: 10.4049/jimmunol.166.3.1983 Downloaded from http://www.jimmunol.org/content/166/3/1983 References This article cites 74 articles, 30 of which you can access for free at: http://www.jimmunol.org/ http://www.jimmunol.org/content/166/3/1983.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists by guest on September 25, 2021 • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2001 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Differential Regulation of the Expression of CD95 Ligand, Receptor Activator of Nuclear Factor-␬B Ligand (RANKL), TNF-Related Apoptosis-Inducing Ligand (TRAIL), and TNF-␣ During T Cell Activation1,2 Ruoxiang Wang,* Liying Zhang,* Xiaoren Zhang,* Jose Moreno,† Xunyi Luo,* Mehrdad Tondravi,† and Yufang Shi3* Members of TNF superfamily are characterized by their ability to inflict apoptosis upon binding to their cognate receptors in a homotrimeric manner. These proteins are expressed on different cell types under various conditions. However, the mechanisms governing the expression of these molecules remain elusive. We have found that the TCR signal can elicit the expression of receptor ␬ ␣ activator of NF- B ligand (RANKL), TNF- , CD95L, and TNF-related apoptosis inducing ligand (TRAIL) in T cell hybridoma Downloaded from A1.1 cells, thus allowing us to examine the expression pattern of these molecules under precisely the same conditions. We have previously reported that CD95L expression requires both protein kinase C (PKC) translocation and Ca2؉ mobilization and is inhibited by cyclosporin A, and dexamethasone. We demonstrate now that activation-induced expression of RANKL is mediated -by Ca2؉ mobilization. PKC activation does not induce RANKL expression nor does it synergize with the Ca2؉ signal. Activation induced RANKL expression is blocked by cyclosporin A, but not by dexamethasone. The expression of TNF, in contrast, is ␣ 2؉ mediated by PKC, but not by Ca . TNF- expression is not inhibited by cyclosporin A, but is sensitive to dexamethasone. A1.1 http://www.jimmunol.org/ cells constitutively express TRAIL at low levels. Stimulation with anti-CD3 leads to an initial reduction and subsequent increase in TRAIL expression. TRAIL induction is not inhibited by cyclosporin A, but highly sensitive to dexamethasone. Therefore, expression of the TNF superfamily genes is regulated by distinct signals. Detailed understanding of the regulatory mechanisms could provide crucial information concerning the role of these molecules in the modulation of the immune system. The Journal of Immunology, 2001, 166: 1983–1990. he TNFR superfamily, a group of type I transmembrane through their cognate receptors (5). Although the biological role of glycoproteins, represents a growing family of receptors some of these ligand-receptor pairs remains obscure, a number of T fundamental for cell growth, differentiation, and apoptosis studies have revealed insights into the mechanisms by which the ex- by guest on September 25, 2021 during development and pathophysiological processes. Several family pression of the TNF superfamily genes is regulated. TNF-␣ is pro- members have been identified in mammalian cells. Each member of duced by a variety of cell types in response to diverse stimuli. The the TNFR family binds to its distinct cognate ligand(s) (1, 2). The regulation of TNF-␣ is best appreciated in response to signals from identification of this family of proteins is based on the shared homol- the TCR ligation. It has been reported that the production of TNF-␣ ogies in their cysteine-rich extracellular ligand-binding domains and in T cell clones depends on Ca2ϩ mobilization induced by TCR stim- intracellular effector (death) domains (3). These receptors transmit ulation (6). This Ca2ϩ-dependent induction of TNF-␣ is blocked by their signals mainly via the interactions of their intracellular domains cyclosporin A (6). In B cells, activation through Ag receptor or via the with intracellular proteins bearing death domain-containing proteins CD40 pathway also induces NF-AT-dependent expression of TNF-␣ (4). The discovery of these domains has shed light on the overall that is blocked by cyclosporin A/FK506, although the promoter bind- architecture of the pathway leading from receptor ligation to the ac- ing site of NF-AT may be different from that in T cells (7–9). More- tivation of signaling cascade and eventually to biological functions over, transfection of a cDNA encoding a constitutively active form of such as apoptosis, differentiation, proliferation, and cell survival. calcineurin is sufficient to activate the TNF-␣ promoter (10). There- Concomitantly, the ligands for these receptors, now referred to as fore, the induction of TNF-␣ expression by the TCR activation signals the members of the TNF superfamily, have been identified as a series in lymphocytes is dependent on Ca2ϩ mobilization and the activation of type II transmembrane glycoproteins that bind to and signal of NF-AT. On the other hand, it has been shown that the TNF-␣ promoter can be activated by phorbol esters (PMA) or LPS in human mononuclear cells, a process dependent on NF-␬B (11–13). Interest- Departments of *Immunology and †Tissue Biology, Jerome H. Holland Laboratory, American Red Cross, Rockville, MD 20855 ingly, the existence of ␬ elements in the promoter of TNF-␣ gene is ␣ Received for publication March 27, 2000. Accepted for publication October 31, 2000. not responsible for PMA- and LPS-induced TNF- production (14). ␣ The costs of publication of this article were defrayed in part by the payment of page TNF- promoter analysis revealed that the PMA-responsive element charges. This article must therefore be hereby marked advertisement in accordance is activated by AP-1 (15, 16). with 18 U.S.C. Section 1734 solely to indicate this fact. Our previous studies have demonstrated that activation-induced 1 This work was supported by National Institutes of Health Grants AI43384, expression of CD95 ligand (CD95L)4 requires both protein kinase CA76492, and AR44089. 2 This is publication number 107 of the Department of Immunology, Holland Labo- ratory of the American Red Cross. 4 Abbreviations used in this paper: CD95L, CD95 ligand; AICD, activation-induced 3 Address correspondence and reprint requests to Dr. Yufang Shi, Department of cell death; FasL, Fas ligand; OPG, osteoprotegerin; PKC, protein kinase C; RANKL, Immunology, Jerome H. Holland Laboratory of the American Red Cross, 15601 receptor activator of NF-␬B ligand; TMB-8, 8-(diethylamino)-octyl-3,4,5-trimethoxy- Crabbs Branch Way, Rockville, MD 20855. E-mail address: [email protected] benzoate hydrochloride; TRAIL, TNF-related apoptosis-inducing ligand. Copyright © 2001 by The American Association of Immunologists 0022-1767/01/$02.00 1984 DIFFERENTIAL REGULATION OF GENES IN THE TNF SUPERFAMILY 2ϩ C (PKC) translocation and Ca mobilization (17). The expression mM L-glutamine, 50 mM 2-ME, 10% heat-inactivated FBS (Sigma, St. of CD95L can be blocked by either PKC inhibitors or blockers of Louis, MO), and 10 mM gentamicin. Cell cultures were incubated at 37°C 2ϩ in humidified atmosphere with 5% CO /95% air. Ca mobilization (17). The same finding has also been reported 2 by others (18). A recent study has demonstrated that the expression Reagents of CD95L is mediated by PKC␪ (19). In addition, it has also been Ab to CD3 was produced by a hamster B cell hybridoma, 145-2C11⑀ (from shown that CD95L expression is sensitive to cyclosporin A, reti- Dr. Jeffery Bluestone, University of Chicago, Chicago, IL). PMA, N-(2- noic acid, and dexamethasone (17, 20–22). Promoter analysis has guanidinoethyl)-5-isoquinolinesulfonamide (HA 1004), 1-(5-isoquinoline- revealed that the activation of CD95L transcription requires sulfonyl)-2-methylpiperazine (H7), GO¨ 6976, bisindolylmaleimide I, Cal- NF-␬B (23, 24), NF-AT (25, 26), Nurr-77 (27), ALG-4 (28), Egr-3 phostin C, and ionomycin were purchased from Calbiochem (San Diego, CA). An inhibitor for intracellular Ca2ϩ mobilization, 8-(diethylamino)- (29), and AP-1 (30). Several of these transcription factors have octyl-3,4,5-trimethoxybenzoate hydrochloride (TMB-8), dexamethasone, been shown to directly regulate CD95L transcription in response to and retinoic acid were obtained from Sigma. Cyclosporine was a gift from TCR activation. In addition, we have reported that activation-in- Novartis Pharmaceuticals (East Hanover, NJ). FR901228 was obtained duced CD95L expression requires the participation of the protoon- from Fujisawa Pharmaceutical Company (Osaka, Japan). rDR5 was kindly cogene c-myc (31). provided by Dr. Youhai Chen (University of Pennsylvania, Philadelphia, ␬ PA). Anti-murine CD95L, MFL3, was purchased from PharMingen (La Receptor activator of NF- B ligand (RANKL)/TNF-related ac- Jolla, CA), and rOPG was from R&D Systems (Minneapolis, MN). Other tivation-induced cytokine (TRANCE) has been shown to be ex- chemicals used were the purest grade available from Sigma. pressed in T cells and other cell types (32–34). Its major function TCR activation has been attributed to the regulation of skeletal homeostasis acting in concert with its decoy receptor osteoprotegerin (OPG) (35, 36).