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Cells by Rat and Human Type II Alveolar Epithelial Inducible Expression of the α1-Acid Glycoprotein by Rat and Human Type II Alveolar Epithelial Cells This information is current as Bruno Crestani, Corinne Rolland, Bernard Lardeux, Thierry of October 1, 2021. Fournier, Dominique Bernuau, Christian Poüs, Christiane Vissuzaine, Lin Li and Michel Aubier J Immunol 1998; 160:4596-4605; ; http://www.jimmunol.org/content/160/9/4596 Downloaded from References This article cites 41 articles, 17 of which you can access for free at: http://www.jimmunol.org/content/160/9/4596.full#ref-list-1 Why The JI? Submit online. http://www.jimmunol.org/ • 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 by guest on October 1, 2021 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 © 1998 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. a Inducible Expression of the 1-Acid Glycoprotein by Rat and Human Type II Alveolar Epithelial Cells1 Bruno Crestani,2* Corinne Rolland,* Bernard Lardeux,† Thierry Fournier,* Dominique Bernuau,† Christian Pou¨s,‡ Christiane Vissuzaine,§ Lin Li,* and Michel Aubier* a 1-Acid glycoprotein (AGP) is a major acute phase protein in rat and human. AGP has important immunomodulatory functions that are potentially important for pulmonary inflammatory response. The liver is the main tissue for AGP synthesis in the organism, but the expression of AGP in the rat lung has not been investigated. We show that AGP mRNA was induced in the lung of dexamethasone-, turpentine-, or LPS-treated rats, whereas AGP mRNA was not detected in the lung of control rats. In the lung of animals treated intratracheally with LPS, in situ hybridization showed that AGP gene expression was restricted to cells located in the corners of the alveolus, consistent with an alveolar type II (ATII) cell localization. The inducible expression of the AGP gene was confirmed in vitro with SV40 T2 cells and rat ATII cells in primary culture: maximal expression required the presence of Downloaded from dexamethasone. IL-1 and the conditioned medium of alveolar macrophages acted synergistically with dexamethasone. Rat ATII cells secreted immunoreactive AGP in vitro when stimulated with dexamethasone or with a combination of dexamethasone and the conditioned medium of alveolar macrophages. In vivo, in the human lung, we detected immunoreactive AGP in hyperplastic ATII cells, whereas we did not detect AGP in the normal lung. We conclude that AGP is expressed in the lung in cases of inflammation and that ATII cells are the main source of AGP in the lung. The Journal of Immunology, 1998, 160: 4596–4605. http://www.jimmunol.org/ he host response to tissue injury, i.e., the “acute phase tion by monocytes/macrophages (5), and increases the secretion of response,” is a highly coordinated series of physiologic an IL-1 inhibitor by murine macrophages, most probably the IL-1 T reactions involving almost every major organ system. receptor antagonist (IL-1Ra) (6, 7). In vivo, AGP protects mice Marked changes in concentrations of plasma glycoproteins termed from TNF-a-induced lethality (8). The marked increase in plasma “acute phase plasma proteins” occur in the course of the acute AGP concentration in the course of the acute phase response could phase of inflammation (1). The latter result from changes in the therefore act as a form of negative feedback aimed at limiting the expression of specific genes in the liver and are the consequences extent of the inflammatory reaction and its possible deleterious of the action of inflammatory mediators and hormones such as consequences. A local expression of AGP, at the site of the initial glucocorticosteroids on hepatocytes (1). acute phase reaction, could also serve as a protection against the by guest on October 1, 2021 a 3 The 1-acid glycoprotein (orosomucoid, AGP) is a typical deleterious effect of inflammation. This could be particularly im- acute phase plasma protein in humans, rats, mice, and other species portant in the alveolar space, an essential part of the lung, where (2). AGP is a single polypeptide with a molecular mass of 23 kDa gas exchange takes place. Indeed, the integrity of this very delicate plus three to five highly sialylated carbohydrate side chains, the and specialized structure is essential for the maintenance of the ; ; latter accounting for 45% of its total mass of 45 kDa (3). function of the organ. Thus, any inflammatory reaction developing b a IL-1 , TNF- , and IL-6 have been shown to be the key cytokines in the lung must be tightly controlled to preserve the structure of controlling the hepatocyte expression of AGP (1). Glucocorticoids the alveolar space. b stimulate AGP expression and act synergistically with IL-1 , There is a growing body of evidence that the acute phase re- a TNF- , and IL-6 to induce AGP expression by hepatocytes (1). sponse may take place in extrahepatic cell types, notably epithelial AGP has been shown to act in vitro and in vivo as an immu- cells, and may be regulated by cytokines, as observed in hepato- nomodulatory molecule. In vitro, AGP inhibits polymorphonuclear cytes (9–12). Extrahepatic synthesis of AGP has been detected in neutrophil activation (4), modulates LPS-induced cytokine secre- vivo in the mouse kidney and the pregnant rat uterus (9, 13) and in the human prostate and myocardium (14, 15). However, AGP ex- *Institut National de la Sante´ et de la Recherche Me´dicale (INSERM) U408 and pression in the lung, either normal or inflamed, has never been †INSERM U327, Faculte´deMe´decine Xavier Bichat, and ‡Laboratoire de Biochimie A and §Laboratoire d’Anatomie-Pathologique, Hoˆpital Bichat, Assistance Publique- detected. Hoˆpitaux de Paris, Paris, France In view of the important immunomodulatory properties of AGP, Received for publication April 22, 1997. Accepted for publication January 8, 1998. we asked whether AGP is expressed in the rat lung in the course The costs of publication of this article were defrayed in part by the payment of page of different types of inflammatory response, and if so, how its charges. This article must therefore be hereby marked advertisement in accordance expression is controlled. We used two models of inflammation: 1) with 18 U.S.C. Section 1734 solely to indicate this fact. LPS administration (either intratracheal or i.p.) to mimic infection 1 This work was supported by a grant from Zeneca Pharma, Cergy, France. and 2) s.c. turpentine injection to induce a nonseptic localized 2 Address correspondence and reprint requests to Docteur Bruno Crestani, Unite´de inflammation. In this article, we show for the first time that AGP Pneumologie, Hoˆpital Bichat, 46 rue Henri Huchard, 75018 Paris, France. E-mail address: [email protected] mRNA is expressed in the rat lung in cases of inflammation or 3 a steroid administration, that alveolar type II cells (ATII cells) are Abbreviations used in this paper: AGP, 1-acid glycoprotein; AM-CM, alveolar macrophage-conditioned medium; ATII cells, type II alveolar cells; rh, recombinant the primary site of AGP expression in the rat lung in vivo, and that human; rm, recombinant murine; GuSCN, guanidine thiocyanate; DMEM, Dulbec- co’s modified Eagle’s medium; GAPDH, glyceraldehyde-phosphate dehydrogenase; rat ATII cells in vitro express the AGP mRNA and secrete immu- RPA, ribonuclease protection assay; nt, nucleotide. noreactive AGP when stimulated with the secretory products of Copyright © 1998 by The American Association of Immunologists 0022-1767/98/$02.00 The Journal of Immunology 4597 alveolar macrophages and dexamethasone. Moreover, we show esterase stain (Sigma). Conditioned medium, consisting of LPS-activated that hyperplastic alveolar epithelial cells express immunoreactive rat alveolar macrophages, was recovered after a 24-h incubation period. AGP in the human lung. Stimulation of AGP gene expression and AGP secretion by rat ATII cells in vitro Materials and Methods Rat ATII cells were used 24 h after isolation. Cells were incubated with Reagents complete DMEM. Rat CM (10% v/v) or cytokines (10 ng/ml) were added 25 Recombinant murine (rm)IL-1, rmTNF-a, and recombinant human for the indicated times in the presence or absence of dexamethasone (10 (rh)IL-6 were purchased from Immugenex (Los Angeles, CA). Escherichia M). In these experiments, we stimulated rat ATII cells with murine recom- coli (strain 026; B6)-derived LPS was obtained from Difco (Detroit, MI). binant cytokines because rat products were not available. These cytokines TEMED, ammonium persulfate, urea, dexamethasone, and turpentine were have previously been shown to be active on rat cells (16, 17). Supernatants from Sigma (La Verpillie`re, France). Transcription reagents were pur- were recovered for the determination of AGP concentration. Cell mono- m 3 chased from Promega (Madison, WI). [a-32P]UTP (400 Ci/mmol) was layers were scrapped in 500 l of PBS and centrifuged (5 min, 12,000 from Amersham (Les Ulis, France). RNase-free DNase I, RNase A, and T1 g, 4°C), and the pellet was solubilized in 5 M GuSCN, 0.1 M EDTA, pH m 3 6 and brewer’s yeast tRNA were supplied by Boehringer (Mannheim, Ger- 7.0 (100 l per 2 10 pneumocytes) (18). The cell lysates were stored at 2 many). Acrylamide/bisacrylamide, phenol, and proteinase K were from 80°C until mRNA analysis.
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