Gorillas with Spondyloarthropathies Express an MHC Class I Molecule with Only Limited Sequence Similarity to HLA-B27 that Binds Peptides with Arginine at P2 This information is current as of September 27, 2021. Julie A. Urvater, Heather Hickman, John L. Dzuris, Kiley Prilliman, Todd M. Allen, Kevin J. Schwartz, David Lorentzen, Clare Shufflebotham, Edward J. Collins, Donald L. Neiffer, Bonnie Raphael, William Hildebrand, Alessandro Sette and David I. Watkins Downloaded from J Immunol 2001; 166:3334-3344; ; doi: 10.4049/jimmunol.166.5.3334 http://www.jimmunol.org/content/166/5/3334 http://www.jimmunol.org/ References This article cites 59 articles, 20 of which you can access for free at: http://www.jimmunol.org/content/166/5/3334.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision by guest on September 27, 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. Gorillas with Spondyloarthropathies Express an MHC Class I Molecule with Only Limited Sequence Similarity to HLA-B27 that Binds Peptides with Arginine at P21 Julie A. Urvater,2* Heather Hickman,† John L. Dzuris,‡ Kiley Prilliman,† Todd M. Allen,* Kevin J. Schwartz,* David Lorentzen,§ Clare Shufflebotham,3* Edward J. Collins,¶ Donald L. Neiffer,4ʈ Bonnie Raphael,§§ William Hildebrand,† Alessandro Sette,‡ and David I. Watkins5*§ The human MHC class I gene, HLA-B27, is a strong risk factor for susceptibility to a group of disorders termed spondyloar- thropathies (SpAs). HLA-B27-transgenic rodents develop SpAs, implicating HLA-B27 in the etiology of these disorders. Several Downloaded from nonhuman primates, including gorillas, develop signs of SpAs indistinguishable from clinical signs of humans with SpAs. To determine whether SpAs in gorillas have a similar HLA-B27-related etiology, we analyzed the MHC class I molecules expressed in four affected gorillas. Gogo-B01, isolated from three of the animals, has only limited similarity to HLA-B27 at the end of the ␣1 domain. It differs by several residues in the B pocket, including differences at positions 45 and 67. However, the molecular model of Gogo-B*0101 is consistent with a requirement for positively charged residues at the second amino acid of peptides bound by the MHC class I molecule. Indeed, the peptide binding motif and sequence of individual ligands eluted from Gogo-B*0101 http://www.jimmunol.org/ demonstrate that, like HLA-B27, this gorilla MHC class I molecule binds peptides with arginine at the second amino acid position of peptides bound by the MHC class I molecule. Furthermore, live cell binding assays show that Gogo-B*0101 can bind HLA-B27 ligands. Therefore, although most gorillas that develop SpAs express an MHC class I molecule with striking differences to HLA-B27, this molecule binds peptides similar to those bound by HLA-B27. The Journal of Immunology, 2001, 166: 3334–3344. he strongest known association between the MHC and these theories, the arthritogenic peptide hypothesis, postulates that disease susceptibility is that of HLA-B27 and inflamma- B27 plays a direct role in pathogenesis by binding an arthritogenic tory spondyloarthropathies (SpAs)6 in humans. Several peptide (or peptides) and presenting it to autoreactive CTLs. Re- T by guest on September 27, 2021 theories have been proposed to explain the role of B27 in the cent transgenic mouse and in vitro studies suggest that the role of development of SpAs and have been reviewed elsewhere (1–3). B27 in the mechanism of disease may be distinct from its primary The primary function of MHC class I molecules is to present en- function as an Ag presentation and CD8ϩ T cell restriction mol- dogenously produced peptides to CTLs. Thus the most intuitive of ecule (4–7). However, the transgenic rat model provides evidence that the specificity of peptides bound to B27 significantly influ- ences the prevalence of arthritis in these animals (8). Thus, *Wisconsin Regional Primate Research Center, University of Wisconsin, Madison, WI 53715; †Department of Microbiology and Immunology, University of Oklahoma whether the disease mechanism involves B27 functioning in its Health Sciences Center, Oklahoma City, OK 73190; ‡Eppimune, San Diego, CA conventional role of peptide binding molecule remains in question. 92121; §University of Wisconsin Histocompatibility Laboratory, Division of Labo- There are several features of B27 that make it unique among ratory Medicine, Department of Pathology and Laboratory Medicine, Madison, WI 53792; ¶Department of Microbiology and Immunology, University of North Carolina, MHC class I molecules. Crystal structures and molecular models Chapel Hill, NC 27599; ʈPittsburgh Zoo, Pittsburgh, PA 15206; and §§Wildlife Health have demonstrated that B27 is unique in its possession of an un- Sciences, Bronx, NY 10460 usually deep B pocket when compared with other MHC class I Received for publication July 24, 2000. Accepted for publication December 11, 2000. molecules (9). Indeed, many groups have now eluted and se- The costs of publication of this article were defrayed in part by the payment of page quenced peptides bound to B27 and found that the peptides con- charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. tained the bulky and positively charged amino acid, arginine, at the 1 This research was supported by a Biomedical Science Award from the Arthritis second position (10–12). Additionally, the combination of amino Foundation and by National Institutes of Health Grant RR00167 (to the Wisconsin acids that make up the B27 B pocket is unique to and conserved in Regional Primate Research Center). all B27 subtypes (13), all of which bind peptides with arginine at 2 Current address: Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue the second position. Of these residues, glutamic acid at position 45 North, Seattle, WA 98109. (E45), and cysteine at position 67 (C67) have been shown to be 3 Current address: Zeneca Pharmaceuticals, Mereside, Alderley Park, Macclesfield, critical for peptide binding, cell surface expression, and CTL rec- Cheshire, SK10 4TG, U.K. 4 ognition (13, 14). C67 has also been implicated in triggering au- Current address: Disney’s Animal Kingdom, P.O. Box 10000, Lake Buena Vista, FL ␤ 32830. toimmunity (15) and in the formation of unique 2-microglobulin ␤ 5 Address correspondence and reprint requests to Dr. David I. Watkins, Wisconsin ( 2m)-free B27 heavy chain homodimers (7). Regional Primate Research Center, University of Wisconsin, 1220 Capitol Court, Although B27 is the primary genetic factor determining suscep- Madison, WI 53715. E-mail address: [email protected] tibility to SpAs, not all B27-positive individuals develop disease; 6 Abbreviations used in this paper: SpAs, spondyloarthropathies; AS, ankylosing this remains one of the mysteries of these disorders. In general, spondylitis; ReA, reactive arthritis; P2, second amino acid of peptides bound by MHC ϳ class I molecule; PBR, peptide binding region; nano-ES-MS/MS, nanoelectrospray 0.2% of the general population will develop ankylosing spon- tandem mass spectrometry. dylitis (AS), whereas 2% of B27-positive individuals will develop Copyright © 2001 by The American Association of Immunologists 0022-1767/01/$02.00 The Journal of Immunology 3335 the disease (16). Although there is no direct data showing what from primers in Lawlor et al. (23). We designed additional locus-specific percentage of B27-positive individuals will develop reactive ar- primer sets that were also used for these animals. These were: 5ЈGGAXHO Ј Ј thritis (ReA), extrapolation from data summarized by Keat indicate (5 -CGGCCTCGAGATGGCGCCCCGAACCCTCSTCCTGCTA-3 ), 3ЈGGAH3 (5Ј-CGGCAAGCTTCACACAAGGCAGCTGTCTCAC that 20–30% of B27-positive individuals that contract Shigella ACTTTA-3Ј), GGCF (5Ј-CKCCCCGAACCCTCA-3Ј), and GGCR (5Ј- should develop ReA (17). Sixty to 80% of ReA patients are B27- AGGCTTTACAAGYGATGAGAGACT-3Ј). Each primer was used at a ␮ positive, a lower association than is seen in AS where as many as final concentration of 0.25 M. The PCR mixture contained 2 mM MgCl2, 96% of patients are B27-positive. However, this allele is only 50 mM Tris, pH 8.3, and 2.5 U Amplitaq DNA polymerase (Perkin-Elmer ϳ Cetus) in a final volume of 100 ␮l. The reactions were denatured initially present in 7% of Caucasian populations. for 2 min at 94°C followed by 30 cycles of 94°C for 1 min, 60°C for 1 min, Although SpAs are a common phenomenon in nonhuman primates, 72°C for 1.5 min, and a single final extension at 72°C for 10 min. RNA was very little is known about the relationship between disease occurrence not available for Harry, so MHC typing was performed on genomic DNA and expression of MHC class I molecules in these species. Previously, extracted from 200 ␮l of blood using the QIAmp Blood kit (Qiagen, Santa ␮ other groups described AS in the gorilla, Beta (18), and post shigel- Clarita, CA). DNA (175 ng) was amplified in a 50- l reaction using the Invitrogen PCR optimizer kit, Buffer F (Invitrogen, Carlsbad, CA) with the losis ReA in Holli and Husani (19). Another gorilla, Harry, also de- MHC class I generic primer NA1STAR1 (5Ј-GCGAATTCGCTCY veloped ReA (D. Neiffer, manuscript in preparation). By examining CACTCCWTGARGTATTTC-3Ј) and the B friendly primer 3ЈGGBA2 skeletal remains, Rothschild and Woods have also observed SpAs in (5Ј-GTCCGCCGCGGTCCAGGAGCT-3Ј) at a final concentration of 20% of gorillas (20).