CD36 or αvβ3 and αvβ5 Integrins Are Not Essential for MHC Class I Cross-Presentation of Cell-Associated Antigen by CD8 α+ Murine Dendritic Cells This information is current as of September 25, 2021. Oliver Schulz, Daniel J. Pennington, Kairbaan Hodivala-Dilke, Maria Febbraio and Caetano Reis e Sousa J Immunol 2002; 168:6057-6065; ; doi: 10.4049/jimmunol.168.12.6057 http://www.jimmunol.org/content/168/12/6057 Downloaded from References This article cites 61 articles, 30 of which you can access for free at: http://www.jimmunol.org/content/168/12/6057.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 25, 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 © 2002 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology ␣ ␤ ␣ ␤ CD36 or v 3 and v 5 Integrins Are Not Essential for MHC Class I Cross-Presentation of Cell-Associated Antigen by CD8␣؉ Murine Dendritic Cells1 Oliver Schulz,* Daniel J. Pennington,† Kairbaan Hodivala-Dilke,‡ Maria Febbraio,§ and Caetano Reis e Sousa2* Cross-presentation of cell-associated Ag is thought to involve receptor-mediated uptake of apoptotic cells by dendritic cells (DC), ␣ ␤ ␣ ␤ and studies with human DC strongly implicate the endocytic receptor CD36 and the integrins v 3 and/or v 5 in this process. In the mouse, cross-presentation was recently shown to be a function of CD8␣؉ DC. Here we report that CD36 is expressed on CD8␣؉, but not on CD8␣؊, DC. To address the role of CD36 in cross-presentation we compared CD36؊/؊ and CD36؉/؉ H-2b DC for their ability to stimulate naive OT-1 T cells specific for OVA plus H-2Kb in the presence of OVA-loaded MHC-mismatched Downloaded from splenocytes as a source of cell-associated Ag for cross-presentation. Surprisingly, no difference was seen between CD36؊/؊ and CD36؉/؉ CD8␣؉ DC in their ability to cross-present cell-associated OVA or to capture OVA-bearing cells. Furthermore, the proliferation of CFSE-labeled OT-1 cells in response to OVA cross-presentation in vivo was normal in CD36؊/؊ bone marrow ␤ chimeras, also arguing against a necessary role for CD36 in cross-presentation by DC or other APC. DC doubly deficient for 3 ␤ and 5 integrins were similarly unimpaired in their ability to cross-present OVA-bearing cells in vitro. These data demonstrate ␤ ␤ http://www.jimmunol.org/ that in the mouse, receptors other than CD36 or 3 and 5 integrins can support the specialized cross-presenting function of .CD8␣؉ DC. The Journal of Immunology, 2002, 168: 6057–6065 n most cells, MHC class I molecules present peptides derived tutively in vivo and is thought to be an important component in the from cytosolic Ags and have only limited ability to present maintenance of peripheral CD8ϩ T cell tolerance (5, 6). To reflect I exogenous Ags acquired from the external milieu by endo- its dual role in immunity and tolerance, MHC presentation of ex- cytosis (1). The bias of the MHC class I presentation pathway for ogenous cell-associated Ag has been termed cross-presentation (5). endogenous Ags has presumably evolved to ensure that the cyto- The existence of a specialized cross-presenting APC was pos- toxic activity of CD8ϩ T cells remains focused on infected targets tulated in 1987 (7), but its identity remained elusive until recently by guest on September 25, 2021 and spares uninfected cells that may have passively acquired despite many attempts to reconstitute cross-presentation in vitro. pathogen Ags from the environment. However, if absolute, the Several reports suggested that both macrophages (M␾)3 and B inability to present exogenous Ags on MHC class I would prevent cells could present soluble exogenous Ags on MHC class I, but this CTL responses against viruses that do not enter APC or against was only seen in situations in which very large amounts of Ag parasites that are confined to the endocytic compartment. In fact, a were delivered to the cells or taken up via specific Ig, implying that large body of evidence suggests that MHC class I presentation of they were unlikely to constitute an efficient cross-presenting APC exogenous Ags can occur in vivo. Twenty-five years ago, Bevan in vivo (8–13). In contrast to M␾ and B cells, dendritic cells (DC) (2, 3) showed the existence of a cross-priming pathway in mice were shown to be able to efficiently present soluble exogenous Ags allowing the induction of CTL responses against minor histocom- on MHC class I, particularly upon macropinocytic uptake or after patibility Ags carried in a cell-based inoculum. Since then, the targeting to Fc receptors (14–16), and efficient MHC class I cross- general importance of cross-priming has become apparent with the presentation of cell-associated Ags was reconstituted in vitro using realization that it is an integral part of the immune response to human DC that had been fed influenza-infected monocytes (17). many tumors, viruses, allografts, and intracellular pathogens (4). DC can also cross-present cellular Ags on MHC class II (18) and In addition, MHC class I presentation of cell-associated Ags by are sufficient for the induction of CD8ϩ T cell cross-tolerance in bone marrow-derived APC has been shown to take place consti- vivo (19). Recently, Den Haan and Bevan (20) identified CD8␣ϩ DC as the major APC type in mouse spleen able to cross-present to CD8ϩ T cells OVA derived from a cell-based inoculum. The *Immunobiology Laboratory, †Lymphocyte Molecular Biology Laboratory, and ‡Cell same DC subset also excels in presenting soluble OVA protein on Adhesion and Disease Laboratory, Cancer Research UK, London Research Institute, MHC class I (21). Together, these observations suggest that DC London, United Kingdom; and §Division of Hematology and Medical Oncology, Department of Medicine, Weill Medical College of Cornell University, New York, might constitute the primary cross-presenting APC in vivo. NY 10021 Nevertheless, the mechanisms underlying cross-presentation by Received for publication January 22, 2002. Accepted for publication March 21, 2002. DC remain poorly understood. Since most instances of cross-pre- The costs of publication of this article were defrayed in part by the payment of page sentation involve cell-based inocula, one hypothesis is that DC charges. This article must therefore be hereby marked advertisement in accordance phagocytose apoptotic cells in the inoculum and re-present the with 18 U.S.C. Section 1734 solely to indicate this fact. acquired cellular Ags on MHC class I and MHC class II. Support 1 This work was supported by Cancer Research UK. 2 Address correspondence and reprint requests to Dr. Caetano Reis e Sousa, Immu- nobiology Laboratory, Cancer Research UK, London Research Institute, Lincoln’s Inn Fields Laboratories, 44 Lincoln’s Inn Fields, London, U.K. WC2A 3PX. E-mail 3 Abbreviations used in this paper: M␾, macrophage; DC, dendritic cell; DN, double address: [email protected] negative; PS, phosphatidylserine; RAG, recombinase-activating gene. Copyright © 2002 by The American Association of Immunologists, Inc. 0022-1767/02/$02.00 ␣ ␤ ␣ ␤ 6058 DC CROSS-PRESENTATION IN THE ABSENCE OF CD36 OR v 3 AND v 5 for this idea comes from work showing that immature human DC hierarchical gates: a scatter gate around live cells, a histogram gate on capable of phagocytosing apoptotic cells are better at cross-pre- CD11cbright cells, followed by gates on three distinct DC subsets defined senting cell-associated Ags than mature cells that have lost phago- using a CD4 vs CD8 dot plot. CD4, CD8, and double-negative (DN) DC populations from CD45.2/CD45.1 mixed bone marrow chimeras were fur- cytic ability (22). Much interest has therefore focused on which ther split into recipient-derived (CD36ϩ) and donor-derived (CD36Ϫ) sub- receptors mediate the uptake of apoptotic cells by DC. Albert et al. sets by gating on CD45.1ϩ and CD45.1Ϫ populations on a CD45.1 (22) used blocking Abs to show that two of these receptors in histogram. Ϫ/Ϫ human DC are CD36 and ␣ ␤ . An alternative integrin, ␣ ␤ , has OT-1 T cells were isolated from lymph nodes of OT-1 ϫ RAG-1 v 5 v 3 mice and depleted of APC by negative selection using magnetic beads. also been implicated in apoptotic cell uptake by human DC (23). Briefly, cells were stained with a mixture of biotinylated mAbs including ␾ ␣ ␤ In contrast, M do not appear to use v 5, leading to the sugges- anti-Fc␥R, anti-CD4, anti-CD11c, anti-Gr-1, and anti-B220, washed, and tion that the receptors used by different phagocytes for uptake of incubated with streptavidin beads (Miltenyi Biotec, Bisley, U.K.). Labeled apoptotic cells are somehow responsible for the difference in the cells were removed on a MACS depletion column, and the flow-through fate of the internalized Ag: cross-presentation in DC vs degrada- fraction was collected, representing unlabeled, APC-depleted OT-1 T cells. ␾ ␣ ␤ tion in M (22). However, the direct involvement of CD36, v 3, Ab staining and flow cytometry ␣ ␤ and v 5 in cross-presentation has never been tested.