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FCRL4 Is an Fc Receptor for Systemic Iga, but Not Mucosal Secretory Iga Yanling Liu, Sofiya Goroshko, Leslie Y

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FCRL4 Is an Fc Receptor for Systemic Iga, but Not Mucosal Secretory Iga Yanling Liu, Sofiya Goroshko, Leslie Y FCRL4 Is an Fc Receptor for Systemic IgA, but Not Mucosal Secretory IgA Yanling Liu, Sofiya Goroshko, Leslie Y. T. Leung, Shilan Dong, Srijit Khan, Paolo Campisi, Evan J. Propst, Nikolaus This information is current as E. Wolter, Eyal Grunebaum and Götz R. A. Ehrhardt of September 28, 2021. J Immunol published online 8 June 2020 http://www.jimmunol.org/content/early/2020/06/05/jimmun ol.2000293 Downloaded from Supplementary http://www.jimmunol.org/content/suppl/2020/06/05/jimmunol.200029 Material 3.DCSupplemental 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 28, 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 © 2020 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Published June 8, 2020, doi:10.4049/jimmunol.2000293 The Journal of Immunology FCRL4 Is an Fc Receptor for Systemic IgA, but Not Mucosal Secretory IgA Yanling Liu,* Sofiya Goroshko,* Leslie Y. T. Leung,* Shilan Dong,* Srijit Khan,* Paolo Campisi,† Evan J. Propst,† Nikolaus E. Wolter,† Eyal Grunebaum,‡ and Go¨tz R. A. Ehrhardt* Fc receptor–like (FCRL) 4 is an immunoregulatory receptor expressed on a subpopulation of human memory B cells of mucosa- associated lymphoid tissue. Fc receptor function of FCRL4 was demonstrated by binding of IgA to FCRL4 following heat aggregation of the Ig. In this study, we demonstrate that FCRL4 recognizes J chain–linked systemic IgA in the absence of heat aggregation. We further demonstrate that mucosal secretory IgA is not recognized by FCRL4 and that systemic IgA binding can be competitively inhibited by recombinant secretory component protein. Finally, we provide evidence that primary FCRL4- bearing human memory B cells are constitutively bound to IgA. Our study provides a mechanism for the negative regulatory Downloaded from activity of FCRL4 on AgR-mediated B cell activation. The Journal of Immunology, 2020, 205: 000–000. he immunoregulatory receptor, Fc receptor–like (FCRL) composed of three to nine Ig domains with significant sequence ho- 4, is selectively expressed on a morphologically and func- mology to those of classical FcgRandFcεR (3, 13). Key observations tionally distinct subpopulation of human memory B cells from Wilson et al. (14) demonstrated binding of FCRL4 to heat- T http://www.jimmunol.org/ (Bmem) (1–3). This immunoregulatory activity is mediated via aggregated IgA, but not heat-aggregated IgG. Heat aggregation of phosphorylation of ITIM in the intracellular domain and subsequent Igs as a means of providing avidity and simulating immune complex recruitment of the SHP-1 and SHP-2 tyrosine phosphatases (4–6). formation suggests low-affinity binding of IgA to FCRL4. IgA exists Recent studies showed that the Ab repertoire encoded by FCRL4+ in the two isotypes IgA1 and IgA2, both of which can be detected in Bmem contained reduced levels of somatic mutations and displayed monomeric and polymeric isoforms. In serum, IgA levels are lower increased reactivity to commensal microbiota compared with the than those of IgG, and IgA1 levels exceed those of the IgA2 isotype. FCLR42 counterparts (7). In healthy individuals, FCRL4+ Bmem Although both monomeric and polymeric IgA are present, mono- are restricted to sites of the mucosa (1, 8, 9). However, in the context meric IgA is the prevalent isoform, accounting for ∼90% of total of autoimmune disorders and chronic infectious diseases, FCRL4+ circulating IgA (15–17). In contrast, polymeric IgA is the dominant Ig by guest on September 28, 2021 Bmem were detected in circulation (10–12). isotype in humans at sites of the mucosa where it is produced by In humans, the FCRL family consists of six type I transmembrane lamina propria plasma cells and transported across epithelial barriers receptors, the first five of which are predominantly expressed on to the luminal sites of the gastrointestinal or respiratory tract. On the various B lineage cell populations. The extracellular domains are basolateral side of the epithelium, dimeric J chain–linked IgA binds to the polymeric IgR, the extracellular domain of which is cleaved following transcytosis and remains attached to the Ab as a secretory *Department of Immunology, University of Toronto, Toronto, Ontario M5S 1A8, Canada; †Department of Otolaryngology – Head and Neck Surgery, Hospital for component (SC). The transported IgA in complex with the SC is Sick Children, University of Toronto, Toronto, Ontario M5G 1X8, Canada; and termed secretory IgA (sIgA) (18, 19). All three forms of IgA, mo- ‡ Division of Immunology and Allergy, Hospital for Sick Children, Toronto, Ontario nomeric IgA, dimeric IgA, and sIgA, are recognized by the classical M5G 1X8, Canada CD89 IgAR, although CD89 binding of dimeric IgA is stronger than ORCIDs: 0000-0001-6793-4498 (S.G.); 0000-0002-4883-582X (L.Y.T.L.); 0000- 0003-0392-4316 (S.D.); 0000-0002-2750-8117 (P.C.); 0000-0002-8013- binding observed for monomeric IgA or sIgA (20). 8530 (E.J.P.); 0000-0002-5868-5191 (G.R.A.E.). In an effort to gain deeper understanding of FCRL4 recognition of Received for publication March 16, 2020. Accepted for publication May 15, 2020. IgA, we determined that FCRL4 readily binds systemic IgA (defined in This work was supported by Canadian Institutes of Health Research Grants MOP- this study as serum IgA or mucosal IgA that are not transported across 12614 and PJT-16216 (to G.R.A.E.). epithelial barriers, in contrast to mucosal sIgA that is transported to the Y.L. designed, conducted, and analyzed experiments. S.G. conducted and analyzed luminal side of mucosal membranes), but not mucosal sIgA, in experiments. L.Y.T.L. analyzed experiments and critically appraised the manuscript. complex with the SC. Furthermore, we provide evidence that FCRL4+ S.D. analyzed experiments and critically appraised the manuscript. S.K. analyzed experiments and critically appraised the manuscript. P.C. provided specimens and primary Bmem are constitutively decorated with J chain–linked IgA, a critically appraised the manuscript. E.J.P. provided specimens and critically ap- feature unique to these mucosal lymphocytes. Constitutive occupancy praised the manuscript. N.E.W. provided specimens and critically appraised the manuscript. E.G. provided specimens and critically appraised the manuscript. G.R.A.E. of FCRL4 with J chain–linked IgA provides an explanation for the designed, conducted, and analyzed experiments and wrote the manuscript. nonresponsiveness of FCRL4-bearing Bmem following in vitro AgR Address correspondence and reprint requests to Dr. Go¨tz R. A. Ehrhardt, University ligation and suggests a, to our knowledge, novel regulatory model for of Toronto, Medical Sciences Building, Room 7316, 1 King’s College Circle, B lineage cells preferentially encoding microbiota-reactive AgR. Toronto, ON M5S 1A8, Canada. E-mail address: [email protected] The online version of this article contains supplemental material. Abbreviations used in this article: Bmem, memory B cell; FCRL, Fc receptor–like; Materials and Methods HA, hemagglutinin; IgA1-J, J chain–containing dimeric IgA1; SC, secretory compo- Abs and reagents nent; sIgA, secretory IgA. Fluorescently labeled Abs to IgG, IgM, IgD, CD3, CD19, and CD38 were Copyright Ó 2020 by The American Association of Immunologists, Inc. 0022-1767/20/$37.50 obtained from BD Biosciences (San Jose, CA). Abs to IgA, IgA1, and www.jimmunol.org/cgi/doi/10.4049/jimmunol.2000293 2 RECOGNITION OF SYSTEMIC IgA BY FCRL4 IgA2 were obtained from SouthernBiotech (Birmingham, AL). Anti–J Statistical analyses chain Abs were purchased from Thermo Fisher Scientific (Waltham, MA), and anti-FCRL4 Abs were purchased from Life Technologies (Carlsbad, Statistical analyses for Ig binding to transfected cells were determined using CA). Colostrum IgA preparations were obtained from Sigma-Aldrich (St. Kruskal–Wallis and Mann–Whitney U tests; tonsillar Ig-binding experi- Louis, MO). Details for all Ab reagents are listed in Supplemental Table I. ments were analyzed using Wilcoxon matched pair tests. Cell lines and primary cells Supplementary material Supplemental Fig. 1 depicts the gating strategy used to assess IgA/IgG HEK293T cells were grown in DMEM, supplemented with 10% FBS and + 2 100 U/ml penicillin/streptomycin. BJAB cells were grown in RPMI 1640 binding to primary human FCRL4 and FCRL4 Bmem. Supplemental supplemented with 10% FBS, and 100 U/ml penicillin/streptomycin. All Fig. 2 illustrates the absence of Ig binding to empty vector control cells. Supplemental Table I lists the Ab reagents used in this study. cells were grown in a humidified atmosphere at 37˚C and 5% CO2. Serum was obtained from healthy volunteers and tonsillar tissue from pediatric patients undergoing tonsillectomy was obtained from the Hospital for Sick Children (Toronto, ON, Canada) with informed consent according to the Results Declaration of Helsinki and the institute’s
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