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Antibodies Through Fcrn Binding Monoclonal Pharmacokinetics Of IgG1 Allotypes Influence the Pharmacokinetics of Therapeutic Monoclonal Antibodies through FcRn Binding This information is current as David Ternant, Christophe Arnoult, Martine Pugnière, of October 2, 2021. Christine Dhommée, Daniel Drocourt, Eric Perouzel, Christophe Passot, Nadine Baroukh, Denis Mulleman, Gérard Tiraby, Hervé Watier, Gilles Paintaud and Valérie Gouilleux-Gruart J Immunol published online 18 December 2015 http://www.jimmunol.org/content/early/2015/12/17/jimmun Downloaded from ol.1501780 Supplementary http://www.jimmunol.org/content/suppl/2015/12/17/jimmunol.150178 http://www.jimmunol.org/ Material 0.DCSupplemental Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists by guest on October 2, 2021 • 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 © 2015 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Published December 18, 2015, doi:10.4049/jimmunol.1501780 The Journal of Immunology IgG1 Allotypes Influence the Pharmacokinetics of Therapeutic Monoclonal Antibodies through FcRn Binding David Ternant,*,†,1 Christophe Arnoult,*,1 Martine Pugnie`re,‡ Christine Dhomme´e,* Daniel Drocourt,x Eric Perouzel,x Christophe Passot,*,{ Nadine Baroukh,* Denis Mulleman,*,‖ Ge´rard Tiraby,x Herve´ Watier,*,{ Gilles Paintaud,*,† and Vale´rie Gouilleux-Gruart*,{ Because IgG1 allotypes might have different half-lives, their influence on infliximab (G1m17,1 allotype) pharmacokinetics was investigated in a group of spondyloarthritis patients. Infliximab was found to have a shorter half-life in patients homozygous for the G1m17,1 allotypes than in those carrying the G1m3 with no G1m1 (G1m3,-1) allotype. Because the neonatal FcR (FcRn) is involved in the pharmacokinetics of mAbs, the interaction of different IgG1 allotypes with FcRn was examined using cellular assays and surface plasmon resonance. G1m17,1 mAbs, such as infliximab and rituximab, were shown to bind more efficiently to Downloaded from FcRn and to be transcytosed better than the G1m3,-1 mAb cetuximab, which explains why infliximab is a better competitor for endogenous IgG1 in G1m3,-1 allotype–bearing patients. A set of four allotype variants of adalimumab (G1m17,1; G1m17,-1; G1m3,1; and G1m3,-1) was also tested for its binding to FcRn, revealing that the G1m3,1 variant, not present in commercial mAbs, binds more efficiently to FcRn and is transcytosed better than the other three variants, all of which are found in therapeutic mAbs. The Journal of Immunology, 2016, 196: 000–000. http://www.jimmunol.org/ he neonatal FcR (FcRn), described by Brambell in 1964, is bution throughout the body (6–10). More recently, FcRn was one of the receptors able to bind IgG via the Fc portion of found to be involved in phagocytosis, Ag presentation, and hu- T the latter (1, 2). However, it is functionally distinguish- moral and antitumoral immune responses, opening new functional able from other FcgRs by its binding to Fc at acidic pH 6 and by areas for this multifaceted molecule (11–15). its affinity for another substrate, albumin. IgG and albumin bind Composed of an a-chain and b2-microglobulin, FcRn is an to two opposite sites on FcRn and, therefore, do not behave as HLA class I homolog devoid of any peptide-binding groove be- competitors (3–5). Ligand transcytosis and recycling are well- tween domains 1 and 2 of the a-chain (2). The amino acids that documented FcRn properties that are responsible for the long are crucial for the interaction between FcRn and IgG or albumin half-life of IgG and albumin in blood, as well as for their distri- were identified by mutation experiments and crystallography by guest on October 2, 2021 studies (16, 17). This led to the isolation of IgG variants that bind better to FcRn while retaining pH dependency (17, 18). Such *Universite´ Franc¸ois Rabelais de Tours, CNRS UMR7292, Tours F-37032, France; variants have potentially high therapeutic value in the context †Laboratoire de Pharmacologie-Toxicologie, Centre Hospitalier Re´gional Universi- taire de Tours, Tours F-37032, France; ‡INSERM, U1194, Institut de Recherche en of the many therapeutic mAbs and other IgG Fc- or albumin- Cance´rologie de Montpellier, Universite´ de Montpellier; Montpellier F-34298, containing biologics that target a wide range of diseases, such France; xInvivoGen, Toulouse F-31400, France; {Laboratoire d’Immunologie, Centre Hospitalier Re´gional Universitaire de Tours, Tours F-37032, France; and ‖Service de as cancer, inflammation, infection, or genetic deficiencies (19–22). Rhumatologie, Centre Hospitalier Re´gional Universitaire de Tours, Tours F-37032, Indeed, the development of mAbs or biologics with prolonged France plasma half-lives would allow less frequent injections. Such 1D.T. and C.A. contributed equally to this work. agents could also compete with endogenous IgG for FcRn ORCIDs: 0000-0003-1783-7002 (N.B.); 0000-0002-2139-4171 (H.W.); 0000-0003- binding, accelerating the catabolism of, for example, patho- 0158-1356 (G.P.). genic autoantibodies. Received for publication August 7, 2015. Accepted for publication November 17, IgGs consist of four subclasses (IgG1 to IgG4), but IgG1 is the 2015. most highly represented among the Ab-based biologics approved This work was supported by a public grant overseen by the French National Research Agency as part of the “Investissements d’Avenir” program (reference: ANR-10- for sale around the world. Apart from the sequence diversity of the LABX-53-01) and by support from the European Regional Development Fund variable domains, Ab constant regions are identical among IgG1, (FEDER Grant Agreement: Presage 4940-37478; OutExFon). Measurement of with the exception of the natural genetic markers (Gms) or infliximab serum concentrations was carried out within the Centre Pilote de Suivi Biologique des Anticorps The´rapeutiques (CePiBAc) platform. CePiBAc is cofinanced polymorphisms called allotypes. The IgG1 H chain may express by the European Union. Europe is committed to supporting the Region Centre with FEDER. G1m1 [or G1m(a)], G1m2 [or G1m(x)], G1m3 [or G1m(f)], and Address correspondence and reprint requests to Dr. Vale´rie Gouilleux-Gruart, Uni- G1m17 [or Gm(z)] as the main allotypes. The allotypes are versite´ Franc¸ois Rabelais de Tours, CNRS UMR7292, 10 Boulevard Tonnelle´, 37032 inherited in a codominant Mendelian way, and various sets of Tours, France. E-mail address: [email protected] combinations are found in African, white, and Mongoloid pop- The online version of this article contains supplemental material. ulations (23, 24). Although the G1m3 and G1m17 allotypes are Abbreviations used in this article: FcRn, neonatal FcR; Gm, genetic marker; G1m-1, absence of G1m1; G1m3,-1, G1m3 with no G1m1; hFcRn, human FcRn; DhFcRn, located at the same position in the CH1 domain, the G1m1 truncated human FcRn; JurkatDhFcRn, DhFcRn-expressing Jurkat; PK, pharmacoki- counterpart corresponding to the absence of G1m1 [sometimes AF488 netics; rituximab , rituximab conjugated to Alexa Fluor 488 fluorescent dye; called non(a) or nG1m1 or G1m-1] in the CH3 domain cannot be SPR, surface plasmon resonance. detected serologically because its sequence is also present in other Copyright Ó 2015 by The American Association of Immunologists, Inc. 0022-1767/15/$30.00 IgG isotypes. Therefore, it is called an isoallotype (26). G1m-1 is www.jimmunol.org/cgi/doi/10.4049/jimmunol.1501780 2 FcRn BINDING OF IgG1 ALLOTYPES typically found in conjunction with the presence of G1m3 in and natalizumab (Biogen). All were purchased from the respective phar- whites (26). Consequently, the main combinations of G1m allo- maceutical companies. Adalimumab variants were developed by InvivoGen types in this population are G1m17,1 and G1m3 with no G1m1 (Toulouse, France) using pFUSE-CHIg plasmids expressing the C region of the H chain of the human IgG1 allotypes. The four possible combinations of (G1m3,-1). The latter is more frequent (0.65–0.85) than G1m17,1 G1m17 and G1m3 allotypes were generated: G1m17,1; G1m17,-1; G1m3,1; (0.15–0.35) (23). and G1m3,-1. These variants bore no other allotype, such as G1m2 or the IgG3 allotypes [G3m15,16 (s,t)] are located within the FcRn G1m27 and G1m28 “supernumerary” allotypes, described in African binding site and are known to influence IgG3 binding to FcRn, populations (33). Consequently, these variants only differ at aa positions 120(K or R) of CH1, 12(D or E) and 14(L or M) of CH3. IgG3 half-life, and transport (27). In contrast, all of the classical G1m allotypes (G1m1, G1m17, and G1m3) are located outside of Cell lines the FcRn binding site. Nevertheless, previous studies using dif- Truncated human FcRn (DhFcRn)-expressing Jurkat (JurkatDFcRn) cells ferent human i.v. Ig preparations administered to immunodeficient were obtained by stable transfection of a Jurkat cell line with a plasmid patients and containing variable amounts of IgG1 allotypes sug- containing the human FcRn (hFcRn) sequence deleted from 99 nt corre- gest a differential IgG1 half-life according to the G1m patient sponding to the 33 aa in the C terminus of the protein (34). Plasmids b allotypes (28). Therefore, this study was undertaken to evaluate containing the hFcRn (full-length) and human 2-microglobulin–coding sequences were also transfected into the MDCKII cell line. The cells were the influence of the IgG1 allotypes exposed on patient IgG on the expanded for 24 h before G418 selection (1 mg/ml for Jurkat cells and pharmacokinetics (PK) of a mAb with a given allotype.
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