Acute-Phase Protein α1-Antitrypsin−−A Novel Regulator of Angiopoietin-like Protein 4 Transcription and Secretion This information is current as Eileen Frenzel, Sabine Wrenger, Stephan Immenschuh, of September 28, 2021. Rembert Koczulla, Ravi Mahadeva, H. Joachim Deeg, Charles A. Dinarello, Tobias Welte, A. Mario Q. Marcondes and Sabina Janciauskiene J Immunol 2014; 192:5354-5362; Prepublished online 23 April 2014; Downloaded from doi: 10.4049/jimmunol.1400378 http://www.jimmunol.org/content/192/11/5354 Supplementary http://www.jimmunol.org/content/suppl/2014/04/23/jimmunol.140037 http://www.jimmunol.org/ Material 8.DCSupplemental References This article cites 56 articles, 25 of which you can access for free at: http://www.jimmunol.org/content/192/11/5354.full#ref-list-1 Why The JI? Submit online. by guest on September 28, 2021 • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • 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 © 2014 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Acute-Phase Protein a1-Antitrypsin—A Novel Regulator of Angiopoietin-like Protein 4 Transcription and Secretion Eileen Frenzel,* Sabine Wrenger,* Stephan Immenschuh,† Rembert Koczulla,‡ Ravi Mahadeva,x H. Joachim Deeg,{,‖ Charles A. Dinarello,# Tobias Welte,* A. Mario Q. Marcondes,{,‖ and Sabina Janciauskiene* The angiopoietin-like protein 4 (angptl4, also known as peroxisome proliferator–activated receptor [PPAR]g–induced angiopoietin- related protein) is a multifunctional protein associated with acute-phase response. The mechanisms accounting for the increase in angptl4 expression are largely unknown. This study shows that human a1-antitrypsin (A1AT) upregulates expression and release of angplt4 in human blood adherent mononuclear cells and in primary human lung microvascular endothelial cells in a concen- tration- and time-dependent manner. Mononuclear cells treated for 1 h with A1AT (from 0.1 to 4 mg/ml) increased mRNA of angptl4 from 2- to 174-fold, respectively, relative to controls. In endothelial cells, the maximal effect on angptl4 expression was Downloaded from achieved at 8 h with 2 mg/ml A1AT (11-fold induction versus controls). In 10 emphysema patients receiving A1AT therapy (Prolastin), plasma angptl4 levels were higher relative to patients without therapy (nanograms per milliliter, mean [95% confi- dence interval] 127.1 [99.5–154.6] versus 76.8 [54.8–98.8], respectively, p = 0.045) and correlated with A1AT levels. The effect of A1AT on angptl4 expression was significantly diminished in cells pretreated with a specific inhibitor of ERK1/2 activation (UO126), irreversible and selective PPARg antagonist (GW9662), or genistein, a ligand for PPARg. GW9662 did not alter the ability of A1AT to induce ERK1/2 phosphorylation, suggesting that PPARg is a critical mediator in the A1AT-driven angptl4 http://www.jimmunol.org/ expression. In contrast, the forced accumulation of HIF-1a, an upregulator of angptl4 expression, enhanced the effect of A1AT. Thus, acute-phase protein A1AT is a physiological regulator of angptl4, another acute-phase protein. The Journal of Immunology, 2014, 192: 5354–5362. ngiopoietin-like protein 4 (angptl4, also known as hepatic angiogenesis-related disorders, such as cardiovascular diseases fibrinogen/angiopoietin-related protein, fasting-induced (cardiac hypertrophy and atherosclerosis) (8–12), ischemia (4, 13), adipose factor, or peroxisome proliferator–activated tumor growth (4, 14, 15), diabetes (16, 17), wound healing (18, A by guest on September 28, 2021 receptor [PPAR]g-induced angiopoietin-related protein) is an 19), and inflammation (5). Angptl4 is also believed to play a role endogenous inhibitor of lipoprotein lipase (1). Human angplt4 is in the regulation of airway remodeling in asthma and chronic expressed at high levels in the placenta, heart, muscle, lung, liver obstructive pulmonary disease (20, 21). Little is known about the (2), adipose tissue (3), endothelial cells (4), and also in monocytes/ regulation of angptl4 in vivo. The expression of angptl4 is induced macrophages (5). Adipose tissue is suspected to be an abundant by ischemic and hypoxic conditions and by treatment with glu- source of serum angptl4 (6). cocorticoids, PPAR agonists, and TGF-b, among others (4, 22– Angptl4 is implicated in the regulation of glucose homeostasis, 27). The overexpression of angptl4 associated with acute-phase insulin sensitivity, and lipid metabolism (7) and is associated with reactions, is suggested to have an anti-inflammatory effect. Ex- perimental studies demonstrate that induction of angptl4 expres- sion reduces atherosclerosis development (28) and protects pulmonary *Department of Respiratory Medicine, Hannover Medical School, 30625 Hannover, Germany; †Institute for Transfusion Medicine, Hannover Medical School, 30625 microvascular endothelial cells against endotoxin-induced injury (21). Hannover, Germany; ‡Division of Pulmonary Diseases, Department of Internal Med- x Hence, angptl4 is a multifunctional protein, with potential significance icine, Philipps-Universita¨t Marburg, 35037 Marburg, Germany; Department of Re- spiratory Medicine, University of Cambridge, Cambridge CB2 0QQ, United for disease pathogenesis. Kingdom; {Department of Medicine, University of Washington, Seattle, WA In this study, we demonstrate that angptl4 expression is regulated ‖ 98195; Clinical Research Division, Fred Hutchinson Cancer Research Center, by another acute-phase protein, namely human a1-antitrypsin Seattle, WA 98109; and #Department of Medicine, University of Colorado Denver, Aurora, CO 80045 (A1AT). A1AT is one of the most abundant acute-phase proteins in Received for publication February 10, 2014. Accepted for publication March 19, the circulation and one of the fastest acting inhibitors of neutrophil 2014. proteases. A1AT is a multifunctional protein involved in the reg- This work was supported by Baxter Healthcare, Hannover Medical School, Deutsches ulation of acute-phase responses (29–33). In recent years, previ- Zentrum fur€ Lungenforschung, and the Cambridge Biomedical Research Centre. ously unrecognized functions of A1AT have been identified (34), Address correspondence and reprint requests to Prof. Dr. Sabina Janciauskiene, Depart- including protective effects against atherosclerosis (35), inhibition ment of Respiratory Medicine, Hannover Medical School, Feodor-Lynen Strasse 23, of cell apoptosis (36, 37), and tumor growth (31). 30625 Hannover, Germany. E-mail address: [email protected] The clinical relevance of A1AT is highlighted in individuals The online version of this article contains supplemental material. with inherited ZZ (Glu342Lys) A1AT deficiency (plasma levels Abbreviations used in this article: A1AT, a1-antitrypsin; angptl4, angiopoietin-like protein 4, DCF, 29,79-dichlorofluorescein; DFOM, deferoxamine mesylate; DMOG, below 0.7 g/l, whereas normal values range between 1 and 2 g/l). dimethyloxalylglycine; HIF-1a, hypoxia-inducible factor-1a; HMVEC-L, primary These individuals are at high risk of developing early onset emphy- human lung microvascular endothelial cell; HSA, human serum albumin; PPAR, sema, liver and pancreatic diseases at any age, and in rare cases pan- peroxisome proliferator–activated receptor; ROS, reactive oxygen species. niculitis and vasculitis (38). Twenty-five years ago augmentation Copyright Ó 2014 by The American Association of Immunologists, Inc. 0022-1767/14/$16.00 therapy with A1AT isolated from pooled human plasma was de- www.jimmunol.org/cgi/doi/10.4049/jimmunol.1400378 The Journal of Immunology 5355 veloped to treat patients with A1AT deficiency–related emphy- Patient plasma samples sema. Therapy with A1AT also modulates or prevents tissue injury Ten of 20 patients (4 PiSZ and 6 PiZZ: one female and nine males, age 45 in experimental animal models of human diseases, including graft- [15.3] y [mean [SD]) received weekly Prolastin (60 mg/kg), whereas 10 versus-host-disease, rheumatoid arthritis, autoimmune diabetes, patients (PiZZ: 5 females and 5 males at age 65 [8.34] y [mean [SD]) had no and renal ischemia–reperfusion injury, among others (39, 40). How- therapy (controls). All participants were nonsmokers. The mean forced ever, despite vast anti-inflammatory properties ascribed to A1AT expiratory volume in 1 s (FEV1%) was 44.4 and 64.8% in augmented and nonaugmented patients, respectively. All included patients had normal (41), the mechanisms of its anti-inflammatory effects remain in- levels of C-reactive protein. Plasma samples were obtained just before and completely understood. Our finding that A1AT regulates angptl4 2 h after therapy with Prolastin and at random times from controls and stored gene expression and release reveals previously unrecognized ac- at 280˚C until use. Participants had signed informed consent as required tivity of this protein. Moreover, in vitro