Critical Role of IL-12 in Dendritic Cell-Induced Differentiation of Naive B Lymphocytes This information is current as Bertrand Dubois, Catherine Massacrier, Béatrice Vanbervliet, of September 29, 2021. Jérome Fayette, Francine Brière, Jacques Banchereau and Christophe Caux J Immunol 1998; 161:2223-2231; ; http://www.jimmunol.org/content/161/5/2223 Downloaded from References This article cites 64 articles, 39 of which you can access for free at: http://www.jimmunol.org/content/161/5/2223.full#ref-list-1 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 29, 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 © 1998 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Critical Role of IL-12 in Dendritic Cell-Induced Differentiation of Naive B Lymphocytes1,2 Bertrand Dubois, Catherine Massacrier, Be´atrice Vanbervliet, Je´rome Fayette, Francine Brie`re, Jacques Banchereau,3 and Christophe Caux4 Dendritic cells (DC) are potent APCs initiating immune responses. In a previous report, we demonstrated that DC directly enhance both proliferation and differentiation of CD40-activated naive and memory B cells. The present study deciphers the molecular mechanisms involved in DC-dependent regulation of B cell responses. Herein, we have identified IL-12 as the mandatory molecule secreted by CD40-activated DC that promote the differentiation of naive B cells into plasma cells secreting high levels of IgM. In fact, IL-12 synergizes with soluble IL-6R a-chain (sgp80), produced by DC, to drive naive B cell differentiation. IL-12 is critical for the differentiation of naive B cells into IgM plasma cells, whereas IL-6R signaling mainly promotes Ig secretion by already differentiated B cells. The differentiation of naive B cells in cocultures of B cells, T cells, and DC is IL-12 dependent, definitely Downloaded from demonstrating that the role of DC in humoral responses is not confined to the activation of T cells and further extending the physiologic relevance of DC/B cell interaction. Finally, this study also identifies differential requirements for DC-dependent naive and memory B cell differentiation, the latter being IL-12 independent. Altogether these results indicate that, in addition to prime T cells toward Th1 development, DC, through the production of IL-12, may also directly signal naive B cell during the initiation of the immune response. The Journal of Immunology, 1998, 161: 2223–2231. http://www.jimmunol.org/ endritic cells (DC)5 are bone marrow-derived leukocytes critical importance as illustrated in the hyper IgM syndrome (15) with potent immunostimulatory properties (1). In the pe- or mice deficient for CD40 or CD40L (16, 17). D riphery, DC such as epidermal Langerhans cells, capture Although studies have underlined the specific role of DC in invading Ag/pathogen and migrate via the lymph stream into humoral responses (4–7), the extent to which DC can directly reg- draining lymph nodes. In the paracortical areas, these DC, called ulate B cell activation requires further analysis. Recent progress in interdigitating cells present processed peptides to naive T cells and the propagation of DC in vitro from precursors (18) and in their initiate the immune response. This T cell activation is followed by purification from blood (19) or tonsils (20, 21) has enabled such cognate interaction between Ag-specific T cells and B cells (2), studies. Recently, several findings have suggested that DC may by guest on September 29, 2021 leading to primary Ab production. The essential role of DC in directly regulate B cell activation during the course of the humoral humoral response has been documented in vitro (3) and in vivo response (22–25). In particular, we showed that DC, generated by (4–7) and is thought to be the consequence of the requirement for culturing cord blood CD341 progenitors for 12 days in presence of DC for naive T cell priming. Once activated, T cells express CD40 GM-CSF 1 TNF-a (26, 27), strongly enhance growth and differ- ligand (CD40L), which in return signal DC to up-regulate costimu- entiation of CD40-activated B cells (22). In the present study, latory molecules (CD54, CD80, CD86) and to secrete cytokines aimed at characterizing the molecules involved in the effects of DC (IL-1, IL-6, IL-8, IL-12, TNF-a) (8, 9). In parallel, CD40L ex- on B cells, we demonstrate that 1) DC-derived IL-12 is mandatory pressing T cells together with cytokines promotes B cell survival in inducing naive, but not memory, B cell differentiation, and that (10), proliferation (11), as well as B cell differentiation and isotype 2) DC strongly enhance Ig production by naive and memory B switching (12, 13). Thus, among the signals involved in DC/T cells by potentiating the IL-6-dependent pathway of B cell differ- cell/B cell cooperation, CD40/CD40L interactions (14) appear of entiation. These findings demonstrate that DC-derived IL-12 can directly signal B cells and thus influence the initiation and subse- quent development of the humoral response. Laboratory for Immunological Research, Schering Plough, Dardilly, France Received for publication January 14, 1998. Accepted for publication May 4, 1998. The costs of publication of this article were defrayed in part by the payment of page Materials and Methods charges. This article must therefore be hereby marked advertisement in accordance Hemopoietic factors, cytokines, Abs, and cell lines with 18 U.S.C. Section 1734 solely to indicate this fact. 3 6 1 rhGM-CSF (specific activity: 2 10 U/mg, Schering-Plough Research Preliminary results were presented at the 4th International Symposium on Dendritic Institute, Kenilworth, NJ) was used at a saturating concentration of 100 Cells in Fundamental and Clinical Immunology, held in Venice, Italy, October 1996. ng/ml (200 U/ml). rhTNF-a (specific activity: 2 3 107 U/mg, Genzyme, 2 B.D. was supported by Foundation Marcel Me´rieux, Lyon, France, and J.F. by Ecole Boston, MA) was used at an optimal concentration of 2.5 ng/ml (50 U/ml). Normale Supe´rieure de Lyon, France. Recombinant human stem cell factor (specific activity 4 3 105 U/mg, R&D 3 Current address: Baylor Institute for Immunology Research, 3500 Gaston Avenue, Systems, Abington, U.K.) was used at an optimal concentration of 25 ng/ P.O. Box 710699, Dallas, Texas 75246. ml. rhIL-2 (specific activity: 3 3 106 U/mg, Amgen, Thousand Oaks, CA) 3 7 3 5 4 Address correspondence and reprints requests to Dr. Christophe Caux, Laboratory was used at 20 U/ml. rhIL-12 (1 10 U/mg) and rhsgp80 (1 10 U/mg) for Immunological Research, Schering-Plough, 27 chemin des Peupliers, BP 11, were purchased from R&D Systems and used at 1 ng/ml and 100 ng/ml, 69571 Dardilly, France. E-mail address: [email protected] respectively. rhIL-6, purified from Escherichia coli by ion-exchange chro- . 5 Abbreviations used in this paper: DC, dendritic cell; CD40L, CD40 ligand; GM- matography ( 95% pure), was obtained from DNAX (Palo Alto, CA) and CSF, granulocyte-macrophage CSF; h, human; SAC, Staphylococcus aureus strain used at 10 ng/ml. Formalinized particles of Staphylococcus aureus strain Cowan I; PE, phycoerythrin; SEA, staphylococcal enterotoxin A; SEB, staphylococ- Cowan I (SAC) were purchased as Pansorbin from Calbiochem-Behring cal enterotoxin B; SAg, superantigen; GCDC, germinal center DC. (La Jolla, CA) and were used at a final concentration of 0.01%. Copyright © 1998 by The American Association of Immunologists 0022-1767/98/$02.00 2224 IL-12-DEPENDENT NAIVE B CELL DIFFERENTIATION The following blocking Abs were used at 10 mg/ml: mouse anti-IL-2 and showed the absence of detectable contaminating T cells throughout the mAb (R&D Systems) and mouse anti-gp80 mAb (Diaclone, Besanc¸on, culture. France, and R&D Systems), goat anti-IL-12 and goat anti-IFN-g Abs For certain experiments, naive B cells (104) were cultured in presence (R&D Systems), mouse anti-IL-12 mAb (IgG1, clone C8.6), kindly pro- of irradiated (4000 rad) allogeneic CD41 T cells (105) and increasing num- vided by A. O’Garra, DNAX, Palo Alto, CA), rat anti-hIL-10R mAb (clone bers of irradiated DC (3,000 rad) with or without superantigens (SEA 1 3F9) and anti-hIL-10 mAb (clone 12G8, kindly provided by K. Moore, SEB, 10 ng/ml each). In certain culture conditions, DC were incubated for DNAX), mouse IgG1, goat IgG, and rabbit IgG isotype control Abs (R&D 1 h with SEA 1 SEB, washed 4 times, irradiated (4000 rad), and used to Systems), mouse anti-CD40 (mAb89) and anti-CD40L (LL48) mAbs (pro- stimulate cocultures of T and B cells. duced in our laboratory). In other experiments, B cells and DC were cultured in separate com- The murine fibroblastic cell line stably transfected with human CD40L partments using transwells (Costar, Wilmington, MA). DC (105) cultured (CD40L L cells) was produced in the laboratory as described earlier (28). in the presence or the absence of CD40 triggering (2.5 3 104 irradiated Mouse fibroblastic L cells transfected with the human CD32/FcgRII (29) CD40L L cells or CD32 L cells used as control) in the lower compartment were used as control L cells.