IFN Priming Is Necessary but Not Sufficient To Turn on a Migratory Dendritic Cell Program in Lupus Monocytes This information is current as Alicia Rodriguez-Pla, Pinakeen Patel, Holden T. Maecker, of September 28, 2021. Jose Rossello-Urgell, Nicole Baldwin, Lynda Bennett, Victoria Cantrell, Jeanine Baisch, Marilynn Punaro, Alisa Gotte, Lorien Nassi, Tracey Wright, Anna Karolina Palucka, Jacques Banchereau and Virginia Pascual J Immunol 2014; 192:5586-5598; Prepublished online 14 Downloaded from May 2014; doi: 10.4049/jimmunol.1301319 http://www.jimmunol.org/content/192/12/5586 http://www.jimmunol.org/ Supplementary http://www.jimmunol.org/content/suppl/2014/05/14/jimmunol.130131 Material 9.DCSupplemental References This article cites 70 articles, 26 of which you can access for free at: http://www.jimmunol.org/content/192/12/5586.full#ref-list-1 Why The JI? Submit online. by guest on September 28, 2021 • 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 *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 © 2014 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology IFN Priming Is Necessary but Not Sufficient To Turn on a Migratory Dendritic Cell Program in Lupus Monocytes Alicia Rodriguez-Pla,*,1 Pinakeen Patel,*,1 Holden T. Maecker,† Jose Rossello-Urgell,* Nicole Baldwin,* Lynda Bennett,* Victoria Cantrell,* Jeanine Baisch,* Marilynn Punaro,‡,x Alisa Gotte,‡,x Lorien Nassi,‡,x Tracey Wright,‡,x Anna Karolina Palucka,* Jacques Banchereau,* and Virginia Pascual*,‡ Blood monocytes from children with systemic lupus erythematosus (SLE) behave similar to dendritic cells (DCs), and SLE serum induces healthy monocytes to differentiate into DCs in a type I IFN–dependent manner. In this study, we found that these monocytes display significant transcriptional changes, including a prominent IFN signature, compared with healthy controls. Few of those changes, however, explain DC function. Exposure to allogeneic T cells in vitro reprograms SLE monocytes to acquire DC Downloaded from phenotype and function, and this correlates with both IFN-inducible (IP10) and proinflammatory cytokine (IL-1b and IL6) expression. Furthermore, we found that both IFN and SLE serum induce the upregulation of CCR7 transcription in these cells. CCR7 protein expression, however, requires a second signal provided by TLR agonists such as LPS. Thus, SLE serum “primes” a subset of monocytes to readily (<24 h) respond to TLR agonists and acquire migratory DC properties. Our findings might explain how microbial infections exacerbate lupus. The Journal of Immunology, 2014, 192: 5586–5598. http://www.jimmunol.org/ atients with systemic lupus erythematosus (SLE) often in vivo full differentiation of monocytes into DC-SIGN/CD209a+ suffer from infections, which represent one of the leading monocyte-derived DCs (Mo-DCs) can be triggered by microbial P causes of morbidity and mortality (1). Both the underlying components, such as LPS. These Mo-DCs rapidly lose expression immune alterations of SLE and the immunosuppressive therapy of monocyte markers such as Gr-1/Ly6C and CD115/c-fms and that patients receive to control disease activity predispose to infec- upregulate TLR4 and CD14. They also acquire DC morphology tions. Infection symptoms mimic exacerbations of SLE and often and localize to the T cell areas of lymph nodes via L-selectin and raise important diagnostic and therapeutic dilemmas. Infections, in CCR7. Through Toll/IL-1R domain–containing adapter inducing turn, trigger SLE disease exacerbations. However, the mechanisms IFN-b signaling, they become powerful Ag-capturing cells and responsible for this association have not been elucidated. APCs (6). The extent to which human monocytes differentiate into by guest on September 28, 2021 Myeloid cells, including neutrophils and monocytes, are part of DCs in vivo remains to be fully determined, although DCs found the first line of defense against infectious agents. In addition to their in inflammatory diseases, such as rheumatoid arthritis (RA) and innate immune functions, monocytes can differentiate into den- tumor inflammatory ascitis, share phenotypic and transcriptional dritic cells (DCs) and therefore link innate and adaptive immunity. characteristics with in vitro–generated Mo-DCs (7). Although monocytes exposed in vitro to cytokines and TLR ligands The blood monocyte compartment is altered in SLE (8), as differentiate into DCs (2, 3), their capacity to differentiate in vivo monocytes from a subset of patients behave similar to myeloid has been formally demonstrated only recently (4–6). In mice, DCs (mDCs) by inducing allogeneic CD4+ T cells to proliferate in vitro. Exposure of normal monocytes to SLE serum results in the generation of DCs, which is dependent on IFN-a (8). Un- † *Baylor Institute for Immunology Research, Dallas, TX 75204; Human Immune abated DC maturation could lead to the activation/expansion of Monitoring Center, Institute for Immunity, Transplantation, and Infection, Stanford University Medical Center, Stanford, CA 94305; ‡Division of Pediatric Rheumatol- autoreactive T cells that have escaped central tolerance, thus x ogy, Texas Scottish Rite Hospital for Children, Dallas, TX 75219; and University of explaining many of the features of the disease (9). SLE serum also Texas Southwestern Medical Center, Dallas, TX 75235 drives healthy monocytes to become DCs with B cell helper 1 A.R.-P. and P.P. contributed equally to this work. capabilities. Thus, these DCs can efficiently stimulate naive and Received for publication May 29, 2013. Accepted for publication April 11, 2014. memory B cells to differentiate into IgG and IgA plasmablasts This work was supported by the Baylor Health Care System Foundation and by resembling those found in the blood of SLE patients (10). National Institutes of Health Grants U19 AI082715, AR054083-01, and P50 055503 (to V.P.). In this study, we sought to further characterize the blood monocyte compartment of SLE patients to identify candidate pathways that The microarray data presented in this article have been submitted to the Gene Ex- pression Omnibus (http://www.ncbi.nlm.nih.gov/geo/) under accession number could explain their potential DC behavior. We also sought to in- GSE46923. vestigate the synergism between SLE serum factors, including type Address correspondence and reprint requests to Dr. Virginia Pascual, Baylor Institute I IFN, and pathogen-associated molecular patterns, such as LPS, for Immunology Research, 3434 Live Oak, Dallas, TX 75204. E-mail address: in the acquisition of a migratory monocyte/DC phenotype that could [email protected] explain the link between infections and SLE exacerbations. The online version of this article contains supplemental material. Abbreviations used in this article: 7-AAD, 7-aminoactinomycin D; BDCA, blood dendritic cell Ag; DC, dendritic cell; mDC, myeloid DC; Mo-DC, monocyte-derived Materials and Methods DC; RA, rheumatoid arthritis; SLE, systemic lupus erythematosus; SLEDAI, SLE Subjects disease activity index. All SLE patients were collected from the Pediatric Rheumatology Clinic at Copyright Ó 2014 by The American Association of Immunologists, Inc. 0022-1767/14/$16.00 Texas Scottish Rite Hospital (Dallas, TX). Samples from 51 SLE patients, www.jimmunol.org/cgi/doi/10.4049/jimmunol.1301319 The Journal of Immunology 5587 including 7 males and 44 females, were studied in this project. The average 22 y). The ethnic breakdown of the healthy donors was 42% white, 29% age of the patients at the day of sample collection was 15 y (range, 8–19 y), Hispanic, 19% African American, and 10% Asian. Some of the flow and the average duration of SLE was 0.69 y (range, 0–2.06 y). The cytometry staining on cultured monocytes was done using monocytes breakdown of the patient ethnicity was 45% Hispanic, 27% African from three adult healthy donors (two males and one female with ages American, 18% white, 4% Asian, and 2% unspecified. Table I depicts the ranging from 31 to 56 y). SLE disease activity index (SLEDAI) and medications for each patient in Whole blood samples were collected from pediatric patients and the study. Thirteen sera from pediatric SLE patients were used for the flow healthydonors usingstandardvenipuncture techniques. Bloodwas drawn cytometry staining on cultured monocytes, most of them in more than one in either EDTA or sodium citrate vacutainer tubes (BD Biosciences, independent experiment. SLE sera were selected based on a high SLEDAI Franklin Lakes, NJ) for the cellular immunology experiments, and the score, absence of immunosuppressive medication, and absence of high- samples were used within 4 h of collection. All blood collection protocols dose prednisone at the moment of blood draw, in addition to absence of i.v. were reviewed and approved by the Institutional Review Boards at the prednisolone bolus administration in the 2 mo previous to