Antigen delivery to early endosomes eliminates the superiority of human blood BDCA3+ dendritic cells at cross presentation

Cohn, Lillian B.; Chatterjee, Bithi; Esselborn, Filipp; Smed‐Sörensen, Anna; Nakamura, Norihiro; Chalouni, Cécile; Lee, Byoung-Chul; Vandlen, Richard; Keler, Tibor; Lauer, Peter; Brockstedt, Dirk G.; Mellman, Ira; Delamarre, Lélia

doi:10.1084/jem.20121251

Cited by 169 publications

(214 citation statements)

References 59 publications

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“…cells without drug treatment or cell permeabilization. Previous studies of Ag delivery via individual DC receptors supported the notion that the amount of Ag delivered does not dictate Ag presentation efficiency (5,6). In the present study, we confirm this with quantitative measurements of internalized Ag load that have not been performed previously.…”

Section: Discussionsupporting

confidence: 90%

“…Although CD8 2 DCs receive a lower Ag load via DEC205 compared with their CD8 + DC counterparts, this supports a large body of evidence that CD8 + DCs possess specialized attributes that enhance the efficiency of MHC I cross-presentation relative to other DC subtypes (36)(37)(38). For human DCs, although there is less specialization between DC subsets for MHC I cross-presentation of soluble Ag (5,28,39,40), BDCA-3 + DCs emerge as superior cross-presenters of Ag that can access the late endosomal compartments (6). This suggests that access of cross-presentation by Ab-targeted vaccination requires selection of a targeted receptor that is expressed by more specialized cross-presenting DC subtypes.…”

Section: Discussionsupporting

confidence: 69%

“…Specific trafficking routes that initiate more efficient MHC I or MHC II Ag presentation have not been strictly defined. For example, DEC205 (CD205), a molecule that traffics to late endosomes (4)(5)(6), is considered a superior receptor for MHC I cross-presentation (7)(8)(9)(10). Alternatively, MHC I cross-presentation is efficiently enhanced by receptors that traffic to early, but not late, endosomes (5,6,11,12).…”

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confidence: 99%

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Criteria for Dendritic Cell Receptor Selection for Efficient Antibody-Targeted Vaccination

Reuter¹,

Panozza²,

Macri³

et al. 2015

The Journal of Immunology

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Ab-targeted vaccination involves targeting a receptor of choice expressed by dendritic cells (DCs) with Ag-coupled Abs. Currently, there is little consensus as to which criteria determine receptor selection to ensure superior Ag presentation and immunity. In this study, we investigated parameters of DC receptor internalization and determined how they impact Ag presentation outcomes. First, using mixed bone marrow chimeras, we established that Ag-targeted, but not nontargeted, DCs are responsible for Ag presentation in settings of Ab-targeted vaccination in vivo. Next, we analyzed parameters of DEC205 (CD205), Clec9A, CD11c, CD11b, and CD40 endocytosis and obtained quantitative measurements of internalization speed, surface turnover, and delivered Ag load. Exploiting these parameters in MHC class I (MHC I) and MHC class II (MHC II) Ag presentation assays, we showed that receptor expression level, proportion of surface turnover, or speed of receptor internalization did not impact MHC I or MHC II Ag presentation efficiency. Furthermore, the Ag load delivered to DCs did not correlate with the efficiency of MHC I or MHC II Ag presentation. In contrast, targeting Ag to CD8+ or CD8− DCs enhanced MHC I or MHC II Ag presentation, respectively. Therefore, receptor expression levels, speed of internalization, and/or the amount of Ag delivered can be excluded as major determinants that dictate Ag presentation efficiency in setting of Ab-targeted vaccination.

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Section: Discussionsupporting

confidence: 90%

Section: Discussionsupporting

confidence: 69%

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confidence: 99%

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Criteria for Dendritic Cell Receptor Selection for Efficient Antibody-Targeted Vaccination

Reuter¹,

Panozza²,

Macri³

et al. 2015

The Journal of Immunology

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“…When compared to CD1c + DCs, CD141 + DCs from blood are more effective at cross-presentation of cellular Ag, immune complexes and Ag targeted to late endosomes such as via the DEC-205 receptor [13][14][15]. For soluble protein Ag, blood CD141 + DCs require activation to become efficient at cross-presentation [13,16], whereas lymphoid tissue and skin CD141 + DCs can also cross-present in the absence of activation [13,16,17].…”

Section: Introductionmentioning

confidence: 99%

“…2016. 46: 329-339 Antigen processing 337 isolated blood or tonsillar CD141 + DCs and CD1c + DCs [15,17], spaciotemporal differences may become apparent during Ag uptake and processing. This may be particularly pertinent for processing of particulate Ag or soluble Ag that traffics to rapidly maturing endosomes, where the kinetics of acidification are crucial [24].…”

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confidence: 99%

Differential uptake and cross‐presentation of soluble and necrotic cell antigen by human DC subsets

et al. 2015

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Cross-presentation is the mechanism by which exogenous Ag is processed for recognition by CD8 + T cells. Murine CD8α + DCs are specialized at cross-presenting soluble and cellular Ag, but in humans this process is poorly characterized. In this study, we examined uptake and cross-presentation of soluble and cellular Ag by human blood CD141 + DCs, the human equivalent of mouse CD8α + DCs, and compared them with human monocyte-derived DCs (MoDCs) and blood CD1c + DC subsets. MoDCs were superior in their capacity to internalize and cross-present soluble protein whereas CD141 + DCs were more efficient at ingesting and cross-presenting cellular Ag. Whilst cross-presentation by CD1c + DCs and CD141 + DCs was dependent on the proteasome, and hence cytosolic translocation, cross-presentation by MoDCs was not. Inhibition of endosomal acidification enhanced cross-presentation by CD1c + DCs and MoDCs but not by CD141 + DCs. These data demonstrate that CD1c + DCs, CD141 + DCs, and MoDCs are capable of crosspresentation; however, they do so via different mechanisms. Moreover, they demonstrate that human CD141 + DCs, like their murine CD8α + DC counterparts, are specialized at cross-presenting cellular Ag, most likely mediated by an enhanced capacity to ingest cellular Ag combined with subtle changes in lysosomal pH during Ag processing and use of the cytosolic pathway.Keywords: Antigen processing r Cross-presentation r Human dendritic cells Additional supporting information may be found in the online version of this article at the publisher's web-site IntroductionDendritic cells (DCs) are professional APCs that are uniquely able to process and present antigen (Ag) to prime naïve T-cell Correspondence: Dr. Kristen J. Radford e-mail: kristen.radford@mater.uq.edu.au responses. DCs in human and mouse can be classified into a number of subsets that vary in location, phenotype, and specialized function [1]. These include (i) inflammatory monocyte-derived DCs (MoDCs) that develop from monocytes and are rapidly * These authors contributed equally to this work.C 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.eji-journal.eu 330 Meng-Chieh Chiang et al. Eur. J. Immunol. 2016. 46: 329-339 recruited to sites of inflammation, (ii) plasmacytoid DCs, which are key producers of type I IFN, and (iii) "classical" or "conventional" DCs (cDCs), which can be further categorized based on location into "lymphoid-resident" and "migratory" DCs [1]. The lymphoidresident DCs capture Ag directly in situ, whereas migratory DCs reside in the peripheral organs (e.g. lung, skin, and gut) where they capture Ag then migrate to lymphoid tissues to share their Ag with other lymphoid-resident DCs, or present Ag directly to T cells. cDCs can be further segregated into two main subsets: (i) the mouse CD11b + cDC subset and human CD1c + DC equivalent; and (ii) the mouse CD8α + lymphoid-resident DC, related mouse CD103 + tissue resident cDCs, and the human equivalent CD141 + DC that can now be collectively defined by coexpression of the C-type lectin-like re...

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Human Dendritic Cell Enrichment and Their Activation of T Cells

Lubin,

Gvili,

Hazan

et al. 2023

Current Protocols

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Dendritic cells (DCs) enable the immune system to mount and modulate precisely targeted responses to various threats across the organism by bridging innate and adaptive immunity. Historically, DCs have been classified as conventional (cDC) and plasmacytoid (pDC). More recently, cDCs were acknowledged as a heterogenous population composed of several subsets. Examining the functional diversity of cDCs in healthy homeostasis and pathology requires a robust experimental pipeline, beginning with an efficient enrichment protocol in preparation for cell sorting. Unfortunately, several commercial DC enrichment kits were developed before the discovery of the more recently described DC populations. Here, we detail two approaches to enrich human blood DCs or certain DC subsets and an in vitro protocol to examine DC stimulation of naïve T cells. The methods employed here overcome many hurdles encountered while enriching human DC subsets. Basic Protocol 1 describes a method that will enrich pDC, Axl Siglec6‐DC (AS‐DC), cDC1, DC2, DC3, monocytes, and human HLA+ cells by crosslinking unwanted cells to erythrocytes. Basic Protocol 2 describes the enrichment of pDC, AS‐DC, cDC1, and DC2 but not DC3 via a highly efficient negative magnetic selection that is valuable in circumstances where DC3 is not required. Finally, Basic Protocol 3 describes a conventional protocol to perform a Mixed leucocyte Reaction (MLR) following the isolation of these DC subsets. These methods detail the advantages and pitfalls when isolating a heterogeneous population of cells. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC.Basic Protocol 1: Human peripheral mononuclear phagocyte enrichmentBasic Protocol 2: DC enrichment of pDC, cDC1, AS‐DC, and DC2 but not DC3Basic Protocol 3: Basic mixed lymphocyte reaction protocol with sorted human DC subsets

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Antigen delivery to early endosomes eliminates the superiority of human blood BDCA3+ dendritic cells at cross presentation

Abstract: Human BDCA3 DCs are superior to BDCA1 DCs at antigen cross presentation when delivered to late endosomes and lysosomes but not when delivered to early endosomes.

Cited by 169 publications

References 59 publications

Criteria for Dendritic Cell Receptor Selection for Efficient Antibody-Targeted Vaccination

Criteria for Dendritic Cell Receptor Selection for Efficient Antibody-Targeted Vaccination

Differential uptake and cross‐presentation of soluble and necrotic cell antigen by human DC subsets

Human Dendritic Cell Enrichment and Their Activation of T Cells

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