Molecular and Immunological Characterization of Arginine Kinase from the Indianmeal Moth, Plodia interpunctella, a Novel Cross-Reactive Invertebrate This information is current as Pan-Allergen of September 25, 2021. Marina Binder, Vera Mahler, Brigitte Hayek, Wolfgang R. Sperr, Matthias Schöller, Sabine Prozell, Gerhard Wiedermann, Peter Valent, Rudolf Valenta and Michael Duchêne Downloaded from J Immunol 2001; 167:5470-5477; ; doi: 10.4049/jimmunol.167.9.5470 http://www.jimmunol.org/content/167/9/5470 http://www.jimmunol.org/ References This article cites 33 articles, 8 of which you can access for free at: http://www.jimmunol.org/content/167/9/5470.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision by guest on September 25, 2021 • 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 © 2001 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Molecular and Immunological Characterization of Arginine Kinase from the Indianmeal Moth, Plodia interpunctella,a Novel Cross-Reactive Invertebrate Pan-Allergen1 Marina Binder,* Vera Mahler,2† Brigitte Hayek,3* Wolfgang R. Sperr,‡ Matthias Scho¨ller,4§ Sabine Prozell,4§ Gerhard Wiedermann,* Peter Valent,‡ Rudolf Valenta,† and Michael Ducheˆne5* IgE recognition of indoor allergens represents a major cause of allergic asthma in atopic individuals. We found that 52 of 102 patients suffering from allergic symptoms indoors contained IgE Abs against allergens from the Indianmeal moth (Plodia inter- punctella), a ubiquitous food pest. Using serum IgE from a moth-sensitized patient we screened an expression cDNA library constructed from P. interpunctella larvae. cDNAs coding for arginine kinase (EC 2.7.3.3), a 40-kDa enzyme commonly occurring Downloaded from in invertebrates that is involved in the storage of such high-energy phosphate bonds as phosphoarginine, were isolated. Recom- binant moth arginine kinase, designated Plo i 1, was expressed in Escherichia coli as a histidine-tagged protein with enzymatic activity, and purified to homogeneity by nickel chelate affinity chromatography. Purified recombinant arginine kinase induced specific basophil histamine release and immediate as well as late-phase skin reactions. It reacted with serum IgE from 13 of the 52 (25%) moth-allergic patients and inhibited the binding of allergic patients’ IgE to an immunologically related 40-kDa allergen http://www.jimmunol.org/ present in house dust mite, cockroach, king prawn, lobster, and mussel. Our results indicate that arginine kinases represent a new class of cross-reactive invertebrate pan-allergens. Recombinant arginine kinase may be used to identify a group of polysensitized indoor allergic patients and for immunotherapy of these individuals. The Journal of Immunology, 2001, 167: 5470–5477. ype I allergic disorders such as rhinoconjunctivitis, atopic et al. (11) characterized various insect extracts by IgE immuno- dermatitis, and bronchial asthma afflict up to 25% of the blotting and demonstrated several IgE-Ags in the clothes moth or T population (1). Arthropods represent more than three silkworm moth. In studies conducted in Japan, a high proportion of quarters of all animal species, and some of those which get into patients with asthma bronchiale (12, 13) or allergic rhinitis (14) close contact with humans are a major allergen source. Whereas were found to react with silkworm moth allergens. by guest on September 25, 2021 the indoor allergens from the house dust mite (2) and cockroach In recent years the Indianmeal moth, Plodia interpunctella, has (3–5), and the allergenic venoms from the vespids (6) have been become a widely spread household and stored product pest studied in detail, much less is known about allergens from moths. throughout the United States and Europe. Its larvae feed on dry That moths could be the causative agent of inhalant allergies had foodstuffs such as nuts, grains, dried fruit, and chocolate (15). Al- been mentioned as early as 1928 by Vaughan (7). Over the years, though it was mentioned as a possible cause of allergies in a re- there have been scattered case reports on bronchial asthma caused view on allergens in mills (16), no detailed studies have been per- by the clothes moth Tineola bisselliella (8) and wax moth Galleria formed whether the Indianmeal moth represents an indoor allergen mellonella (9). Baldo and Panzani (10) and more recently Komase source. We examined a panel of 102 sera from indoor allergic patients and found a high prevalence of IgE reactivity against In- dianmeal moth Ags. One of these IgE-reactive Ags was charac- Divisions of *Specific Prophylaxis and Tropical Medicine and †Immunopathology, terized on the molecular level, and was identified as an arginine Department of Pathophysiology, and Division of ‡Hematology and Hemostaseology, Department of Internal Medicine I, University of Vienna, Vienna, Austria; and §In- kinase by cDNA cloning, demonstration of sequence homology, stitute for Stored-Product Protection, Biological Research Center for Agriculture and and enzymatic activity of the recombinant protein. Finally, we Forestry, Berlin, Germany demonstrate that this allergen has IgE cross-reactive homologs in Received for publication April 9, 2001. Accepted for publication August 31, 2001. several invertebrate species such as mite, cockroach, lobster, king The costs of publication of this article were defrayed in part by the payment of page prawn, and mussel. charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. 1 This study was supported by Grant 8643 from the Jubila¨umsfonds der O¨ sterreich- Materials and Methods ischen Nationalbank and by Grants Y078GEN and F018 from the Austrian Science Patients Fund. Sera from the following groups of patients were tested for the presence of 2 Current address: Department of Dermatology, University of Erlangen-Nuremberg, IgE Abs against moth allergens: 1) patients with type I allergic symptoms Erlangen, Germany. (rhinitis/conjunctivitis, allergic asthma bronchiale) indoors (n ϭ 90, pa- 3 Current address: Division of Allergy, Immunology, and Infectious Diseases, De- tients H1–H90, ages ranging from 17 to 60 years, average age 32 years); 2) partment of Dermatology, University of Vienna, Vienna, Austria. patients with type I allergic symptoms indoors as above plus atopic der- 4 Current address: Biologische Beratung bei Insektenproblemen (BIp), Berlin, matitis (n ϭ 12, patients AH1–AH12, ages from 11 to 47 years, average Germany. age 28 years); 3) control individuals without type I allergies or atopic ϭ 5 Address correspondence and reprint requests to Dr. Michael Ducheˆne, Division of dermatitis (n 10, individuals N1–N10, ages from 26 to 35 years, average Specific Prophylaxis and Tropical Medicine, Department of Pathophysiology, Uni- age 31 years). versity of Vienna, AKH, Wa¨hringer Gu¨rtel 18-20, A-1090 Vienna, Austria. E-mail The diagnosis of type I allergy was based on case history, skin prick address: [email protected] testing, and CAP-RAST (radioallergosorbent test; Pharmacia, Uppsala, Copyright © 2001 by The American Association of Immunologists 0022-1767/01/$02.00 The Journal of Immunology 5471 Sweden) testing using a panel of extracts from indoor (house dust mite, cat This plasmid was checked by DNA sequencing and transformed into E. dander) and outdoor (birch pollen, grass pollen) allergen sources. The di- coli BL21 (DE3) for protein expression. The cells were grown at 37°Cto agnosis of atopic dermatitis was based on the criteria of Hanifin and Rajka an optical density at 600 nm of 0.8. Recombinant protein synthesis was (17). The cDNA library was screened with the serum from patient AH11. induced for3hbyadding isopropylthio ␤-D-galactoside to a final concen- Skin prick tests were performed in patients AH11 and H60. tration of 0.4 mM. The cells were pelleted and lysed by 30-min treatment in buffer L (50 mM NaH2PO4, 300 mM NaCl, 10 mM imidazole, 1 mg IgE reactivity of natural moth, mite, and cockroach extracts as mlϪ1 (w/v) lysozyme, pH 8). Undissolved material was pelleted by 30-min well as the purified recombinant arginine kinase centrifugation at 2000 ϫ g and 4°C. The recombinant protein was then purified by nickel chelate affinity (25) under native conditions using small Preparations from two moth species, house dust mite, and cockroach were spin columns (Qiagen, Hilden, Germany). used to detect specific IgE in patients’ sera. Extracts from the Indianmeal moth P. interpunctella were obtained by homogenizing 25 late-stage larvae per 1 ml of PBS. Reducing gel loading buffer was added 1/1, samples were Measurement of the arginine kinase activity of the recombinant denatured for 10 min at 95°C, and debris was removed by centrifugation in a microcentrifuge (5 min, room temperature, 10,000 ϫ g). In the same way, allergen extracts were prepared from commercial preparations from adult Mediter- Arginine kinase (Enzyme Commission number: EC 2.7.3.3) activity was ranean flour moth (Ephestia kuehniella), house dust mite (Dermatopha- measured by determining the rate of formation of ADP. The ADP is con- goides pteronyssinus), and cockroach (Blattella germanica) obtained from verted back to ATP by pyruvate kinase, and the pyruvate formed is reduced Allergon Pharmacia (Uppsala, Sweden). The extracts or the purified argi- to lactate by lactate dehydrogenase. The rate of consumption of NADH in nine kinase were electrophoresed on preparative 12.5% SDS-polyacryl- this reaction is measured photometrically (26).