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CD30 Shedding from Karpas 299 Lymphoma Cells Is Mediated by TNF- Α-Converting Enzyme

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CD30 Shedding from Karpas 299 Lymphoma Cells Is Mediated by TNF- Α-Converting Enzyme CD30 Shedding from Karpas 299 Lymphoma Cells Is Mediated by TNF- α-Converting Enzyme This information is current as Hinrich P. Hansen, Sebastian Dietrich, Tatiana Kisseleva, of September 25, 2021. Thilo Mokros, Rolf Mentlein, Hans H. Lange, Gillian Murphy and Hilmar Lemke J Immunol 2000; 165:6703-6709; ; doi: 10.4049/jimmunol.165.12.6703 http://www.jimmunol.org/content/165/12/6703 Downloaded from References This article cites 45 articles, 15 of which you can access for free at: http://www.jimmunol.org/content/165/12/6703.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • 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 by guest on September 25, 2021 *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 © 2000 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. CD30 Shedding from Karpas 299 Lymphoma Cells Is Mediated by TNF-␣-Converting Enzyme1 Hinrich P. Hansen,2* Sebastian Dietrich,* Tatiana Kisseleva,3* Thilo Mokros,* Rolf Mentlein,† Hans H. Lange,* Gillian Murphy,‡ and Hilmar Lemke* CD30 is a costimulatory receptor on activated lymphocytes and a number of human lymphoma cells. Specific ligation of mem- brane-bound CD30 or cellular stimulation by PMA results in a metalloproteinase-mediated down-regulation of CD30 and release of its soluble ectodomain (sCD30). In this report, it is demonstrated that PMA-induced CD30 cleavage from Karpas 299 cells was mediated by a membrane-anchored metalloproteinase which was active on intact cells following 3-[(3-cholamidopropyl)dimeth- ylammonio]-1-propanesulfonate extraction of membrane preparations. Moreover, CD30 shedding was blocked by the synthetic hydroxamic acid-based metalloproteinase inhibitor BB-2116 (IC50, 230 nM) and the natural tissue inhibitor of metalloproteinases Downloaded from (TIMP)-3 (IC50, 30 nM), but not by the matrix metalloproteinase inhibitors TIMP-1 and TIMP-2. This inhibition profile is similar to that of the TNF-␣- converting enzyme (TACE) and, indeed, mRNA transcripts of the membrane-bound metalloproteinase- disintegrin TACE could be detected in Karpas 299 cells. The ectodomain of TACE was expressed in bacteria as a GST fusion protein (GST-TACE) which cleaved CD30 from the surface of Karpas 299 cells and concomitantly increased the level of sCD30 in the cell supernatants. Hence, TACE does not only control the release of TNF-␣, but also that of sCD30. The Journal of Immunology, 2000, 165: 6703–6709. http://www.jimmunol.org/ he CD30 is a receptor that has been detected on a subset TNFRs, CD30, CD43, CD44, and the IL-6R, the endoproteolytic of activated T and B cells and a variety of human lym- conversion of membrane-anchored proteins is blocked by broad- T phomas (1, 2). Cloning of its cDNA characterized CD30 spectrum hydroxamate inhibitors of matrix metalloproteinases as a member of the TNFR superfamily (3). The CD30 ligand (MMP). This indicates that MMPs or enzymes related to MMPs (CD30L)4 is expressed on activated T cells, resting B cells, mono- are responsible (8–10). The TNF-␣-converting enzyme (TACE; cytes, and granulocytes and established as a member of the TNF ADAM 17), responsible for the release of TNF-␣ by proteolytic superfamily (4, 5). The CD30-CD30L interaction induces pleio- cleavage of the membrane-associated precursor form (pro-TNF-␣), ϩ tropic biological effects on CD30 cells, including activation, pro- was recently characterized as a membrane-anchored metallopro- by guest on September 25, 2021 liferation, differentiation, and cell death, depending on cell type teinase-disintegrin (ADAM, a metalloproteinase and disintegrin) and accessible other stimuli (6). Soluble CD30 (sCD30) is released (11, 12). Metalloproteinase-disintegrins belong to the metzincin from the cells by proteolytic cleavage of CD30. This CD30 shed- superfamily of metalloproteinases encompassing the physiologi- ϩ ding is enhanced by interaction with the CD30L cells and the cally important families of MMPs, reprolysins and astacins. Due to resulting sCD30 is proposed to reduce the CD30L-dependent ac- their x-ray crystal structure, metalloproteinase-disintegrins are ϩ tivation of CD30 cells (7). closely related to snake venom metalloproteinases (reprolysins) The ectodomains of many membrane proteins can be released and are therefore regarded as members of the reprolysin family of proteolytically from the cell by receptor stimulation or in response metalloproteinases (13). Both snake venom proteinases and met- to protein kinase C activators such as PMA. As demonstrated for alloproteinase-disintegrins exhibit membrane protein sheddase ac- the angiotensin-converting enzyme, the Alzheimer’s amyloid pre- tivity through their catalytic domain (11, 12, 14) and binding to ␣ cursor protein, pro-TNF- , the Fas ligand, CD16, L-selectin, the cellular disintegrin receptors mediated by the disintegrin domain (15–17). Besides pro-TNF-␣, TACE cleaves the amyloid precursor protein (18) and mediates the shedding of L-selectin, the p75 TNFR, and TGF-␣ (16). Other metalloproteinase-disintegrins such Departments of *Biochemistry and †Anatomy, University of Kiel, Kiel, Germany; and ‡School of Biological Science, University of East Anglia, Norwich, United Kingdom as the Drosophila ADAM 10 homologue KUZ and ADAM 9 have Received for publication July 12, 1999. Accepted for publication September 7, 2000. been found to cleave the Notch receptor or the membrane-an- The costs of publication of this article were defrayed in part by the payment of page chored heparin-binding EGF-like growth factor, respectively (19, charges. This article must therefore be hereby marked advertisement in accordance 20). Physiologically, the activity of metalloproteinases is regulated with 18 U.S.C. Section 1734 solely to indicate this fact. by tissue inhibitors of metalloproteinases (TIMPs). Although the 1 This research was supported by a grant from Haensel Stiftung (to H.H.) and a grant activity of MMPs is blocked by TIMP-1, -2, and -3 (21), TACE is from the Bundesministerium fu¨r Bildung und Forschung (to H.L.). only inhibited by TIMP-3 (22). Hence, the shedding of TNF-␣ and 2 Address correspondence and reprint requests to Dr. Hinrich Hansen, Department of Biochemistry, University of Kiel, Olshausenstrasse 40, D-24098 Kiel, Germany. E- L-selectin is inhibited by TIMP-3 and not by TIMP-1 and -2 mail address: [email protected] (23–25). 3 Current address: Department of Microbiology, Columbia University, New York, NY. In the present study, we characterized the CD30-cleaving en- 4 Abbreviations used in this paper: CD30L, CD30 ligand; ADAM, a metalloprotein- zyme found to be membrane-associated and inhibited by the ase and disintegrin; CHAPS, 3-[(3-cholamidopropyl)dimethylammonio]-1-propane- TACE inhibitor TIMP-3. Since TACE-specific mRNA was de- sulfonate; GSSG, oxidized glutathione; MFI, mean fluorescence intensity; MMP, ma- ϩ trix metalloproteinase; phOx, phenyloxazolone; sCD30, soluble CD30; TACE, TNF- tected in CD30 Karpas 299 cells, we describe the bacterial ex- ␣-converting enzyme; TIMP, tissue inhibitor of metalloproteinases. pression of the TACE ectodomain. Its potency to cleave CD30 was Copyright © 2000 by The American Association of Immunologists 0022-1767/00/$02.00 6704 CD30 CLEAVAGE BY TACE compared with that of ADAM 10 and the related snake venom refolded by a 1:100 dilution in either 1) standard refolding buffer (20 mM ␮ metalloproteinase hemorrhargin. Tris-HCl (pH 7.6) containing 200 mM NaCl, 5 mM CaCl2, 100 M ZnCl2, 100 mM arginine, and 0.002% NaN3) or 2) in this buffer which was further supplemented with 1 mM DTT and 3 mM oxidized glutathione (GSSG) Materials and Methods (oxido-shuffling buffer). After incubation for 3 days at 4°C, the crude extract Materials was diluted with the same volume of ice-cold water and the recombinant proteins were purified on glutathione-Sepharose (Amersham Pharmacia Bio- The mAbs Ki-2 (␥1,␬) and Ki-3 (␥2b,␬) were used for detection of cell- tech). The elution of GST-TACE was conducted with 10 mM of reduced bound and sCD30 (26). The anti-2-phenyloxazolone (anti-phOx) Ab BH-1 glutathione in PBS. (␥2b,␬) served as an isotype-matched control in flow cytometry (27). OKT-3 (anti-CD3 mAb) was obtained from American Type Culture Col- Flow cytometry lection (Manassas, VA). The tissue inhibitors of metalloproteinases TIMP-1, TIMP-2, and TIMP-3 and the recombinant ADAM 10 were from Membrane-bound proteins were determined by direct immunofluorescence G.M. (Norwich, U.K.). BB-2116 (N1-(5-acetylamino-1S-methylcarbamoyl- analysis. Cells (3 ϫ 105) were labeled with saturating amounts of FITC- pentyl)-2S-(2,2-dimethyl-propyl)-N4-hydroxy-3R-(4-hydroxy-phenylsulfanyl- conjugated Abs Ki-3-FITC (anti-CD30), anti-CD25-FITC (Dako, Ham- methyl)-succinamide) was a kind gift from British Biotech Pharmaceuti- burg, Germany), or BH1-FITC (anti-phOx) in PBS containing 0.1% BSA cals (Oxford, U.K.). Hemorrhargin from Echis pyramidum laekeyi was and 0.02% sodium azide for 20 min on ice. Cells were analyzed using a donated by Gavin Laing (Liverpool, U.K.). The large cell anaplastic lym- FACScan flow cytometer (Becton Dickinson, Heidelberg, Germany). phoma cell line Karpas 299 was a kind gift from Abraham Karpas (Cam- bridge, U.K.). The cells were cultured at 37°C and 7% CO2 in RPMI 1640 Determination of sCD30 medium supplemented with 10% FCS, 2 mM glutamine, 100 U/ml peni- cillin, and 100 ␮g/ml streptomycin. Flexible 96-well microtiter plates (Integra Bioscience, Fernwald, Ger- many) were coated with 50 ␮l of Ki-2 mAb (50 ␮g/ml) by overnight Stimulation of human lymphocytes incubation at 4°C.
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