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

Human BDCA3+ dendritic cells (DCs), the proposed equivalent to mouse CD8α+ DCs, are widely thought to cross present antigens on MHC class I (MHCI) molecules more efficiently than other DC populations. If true, it is unclear whether this reflects specialization for cross presentation or a generally enhanced ability to present antigens on MHCI. We compared presentation by BDCA3+ DCs with BDCA1+ DCs using a quantitative approach whereby antigens were targeted to distinct intracellular compartments by receptor-mediated internalization. As expected, BDCA3+ DCs were superior at cross presentation of antigens delivered to late endosomes and lysosomes by uptake of anti-DEC205 antibody conjugated to antigen. This difference may reflect a greater efficiency of antigen escape from BDCA3+ DC lysosomes. In contrast, if antigens were delivered to early endosomes through CD40 or CD11c, BDCA1+ DCs were as efficient at cross presentation as BDCA3+ DCs. Because BDCA3+ DCs and BDCA1+ DCs were also equivalent at presenting peptides and endogenously synthesized antigens, BDCA3+ DCs are not likely to possess mechanisms for cross presentation that are specific to this subset. Thus, multiple DC populations may be comparably effective at presenting exogenous antigens to CD8+ T cells as long as the antigen is delivered to early endocytic compartments.

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Internalization and endosomal degradation of receptor-bound antigens regulate the efficiency of cross presentation by human dendritic cells

Blood ◽

10.1182/blood-2012-01-402370 ◽

2012 ◽

Vol 120 (10) ◽

pp. 2011-2020 ◽

Cited By ~ 119

Author(s):

Bithi Chatterjee ◽

Anna Smed-Sörensen ◽

Lillian Cohn ◽

Cécile Chalouni ◽

Richard Vandlen ◽

...

Keyword(s):

Dendritic Cells ◽

Vaccine Development ◽

Mannose Receptor ◽

Antigen Delivery ◽

Cross Presentation ◽

Late Endosomes ◽

Early Endosomes ◽

Antibody Conjugates ◽

Human Dendritic Cells ◽

Endocytic Compartments

Abstract Dendritic cells (DCs) can capture extracellular antigens and load resultant peptides on to MHC class I molecules, a process termed cross presentation. The mechanisms of cross presentation remain incompletely understood, particularly in primary human DCs. One unknown is the extent to which antigen delivery to distinct endocytic compartments determines cross presentation efficiency, possibly by influencing antigen egress to the cytosol. We addressed the problem directly and quantitatively by comparing the cross presentation of identical antigens conjugated with antibodies against different DC receptors that are targeted to early or late endosomes at distinct efficiencies. In human BDCA1+ and monocyte-derived DCs, CD40 and mannose receptor targeted antibody conjugates to early endosomes, whereas DEC205 targeted antigen primarily to late compartments. Surprisingly, the receptor least efficient at internalization, CD40, was the most efficient at cross presentation. This did not reflect DC activation by CD40, but rather its relatively poor uptake or intra-endosomal degradation compared with mannose receptor or DEC205. Thus, although both early and late endosomes appear to support cross presentation in human DCs, internalization efficiency, especially to late compartments, may be a negative predictor of activity when selecting receptors for vaccine development.

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IFN-α enhances cross-presentation in human dendritic cells by modulating antigen survival, endocytic routing, and processing

Blood ◽

10.1182/blood-2011-06-363564 ◽

2012 ◽

Vol 119 (6) ◽

pp. 1407-1417 ◽

Cited By ~ 87

Author(s):

Francesca Spadaro ◽

Caterina Lapenta ◽

Simona Donati ◽

Laura Abalsamo ◽

Vincenzo Barnaba ◽

...

Keyword(s):

T Cells ◽

Dendritic Cells ◽

Mhc Class I ◽

Cd8 T Cells ◽

Tumor Development ◽

Class I ◽

Cross Presentation ◽

The Cross ◽

Human Dendritic Cells

Abstract Cross-presentation allows antigen-presenting cells to present exogenous antigens to CD8+ T cells, playing an essential role in controlling infections and tumor development. IFN-α induces the rapid differentiation of human mono-cytes into dendritic cells, known as IFN-DCs, highly efficient in mediating cross-presentation, as well as the cross-priming of CD8+ T cells. Here, we have investigated the mechanisms underlying the cross-presentation ability of IFN-DCs by studying the intracellular sorting of soluble ovalbumin and nonstructural-3 protein of hepatitis C virus. Our results demonstrate that, independently from the route and mechanism of antigen entry, IFN-DCs are extraordinarily competent in preserving internalized proteins from early degradation and in routing antigens toward the MHC class-I processing pathway, allowing long-lasting, cross-priming capacity. In IFN-DCs, both early and recycling endosomes function as key compartments for the storage of both antigens and MHC-class I molecules and for proteasome- and transporter-associated with Ag processing–dependent auxiliary cross-presentation pathways. Because IFN-DCs closely resemble human DCs naturally occurring in vivo in response to infections and other danger signals, these findings may have important implications for the design of vaccination strategies in neoplastic or chronic infectious diseases.

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Constitutive versus Activation-dependent Cross-Presentation of Immune Complexes by CD8+ and CD8− Dendritic Cells In Vivo

Journal of Experimental Medicine ◽

10.1084/jem.20020295 ◽

2002 ◽

Vol 196 (6) ◽

pp. 817-827 ◽

Cited By ~ 230

Author(s):

Joke M.M. den Haan ◽

Michael J. Bevan

Keyword(s):

T Cells ◽

Dendritic Cells ◽

Mhc Class I ◽

Cd8 T Cells ◽

Immune Complexes ◽

Class I ◽

Cross Presentation ◽

Deficient Mice ◽

Mhc Class I Molecules

Murine splenic dendritic cells (DCs) can be divided into two subsets based on CD8α expression, but the specific role of each subset in stimulation of T cells is largely unknown. An important function of DCs is the ability to take up exogenous antigens and cross-present them in the context of major histocompatibility complex (MHC) class I molecules to CD8+ T cells. We previously demonstrated that, when cell-associated ovalbumin (OVA) is injected into mice, only the CD8+ DC subset cross-presents OVA in the context of MHC class I. In contrast to this selectivity with cell-associated antigen, we show here that both DC subsets isolated from mice injected with OVA/anti-OVA immune complexes (OVA-IC) cross-present OVA to CD8+ T cells. The use of immunoglobulin G Fc receptor (FcγR) common γ-chain–deficient mice revealed that the cross-presentation by CD8− DCs depended on the expression of γ-chain–containing activating FcγRs, whereas cross-presentation by CD8+ DCs was not reduced in γ-chain–deficient mice. These results suggest that although CD8+ DCs constitutively cross-present exogenous antigens in the context of MHC class I molecules, CD8− DCs only do so after activation, such as via ligation of FcγRs. Cross-presentation of immune complexes may play an important role in autoimmune diseases and the therapeutic effect of antitumor antibodies.

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Host ER–parasitophorous vacuole interaction provides a route of entry for antigen cross-presentation in Toxoplasma gondii–infected dendritic cells

Journal of Experimental Medicine ◽

10.1084/jem.20082108 ◽

2009 ◽

Vol 206 (2) ◽

pp. 399-410 ◽

Cited By ~ 115

Author(s):

Romina S. Goldszmid ◽

Isabelle Coppens ◽

Avital Lev ◽

Pat Caspar ◽

Ira Mellman ◽

...

Keyword(s):

T Cells ◽

Dendritic Cells ◽

Toxoplasma Gondii ◽

Mhc Class I ◽

Cd8 T Cells ◽

Parasitophorous Vacuole ◽

Host Cells ◽

Class I ◽

Cross Presentation ◽

Infected Cells

Toxoplasma gondii tachyzoites infect host cells by an active invasion process leading to the formation of a specialized compartment, the parasitophorous vacuole (PV). PVs resist fusion with host cell endosomes and lysosomes and are thus distinct from phagosomes. Because the parasite remains sequestered within the PV, it is unclear how T. gondii–derived antigens (Ag’s) access the major histocompatibility complex (MHC) class I pathway for presentation to CD8+ T cells. We demonstrate that recruitment of host endoplasmic reticulum (hER) to the PV in T. gondii–infected dendritic cells (DCs) directly correlates with cross-priming of CD8+ T cells. Furthermore, we document by immunoelectron microscopy the transfer of hER components into the PV, a process indicative of direct fusion between the two compartments. In strong contrast, no association between hER and phagosomes or Ag presentation activity was observed in DCs containing phagocytosed live or dead parasites. Importantly, cross-presentation of parasite-derived Ag in actively infected cells was blocked when hER retrotranslocation was inhibited, indicating that the hER serves as a conduit for the transport of Ag between the PV and host cytosol. Collectively, these findings demonstrate that pathogen-driven hER–PV interaction can serve as an important mechanism for Ag entry into the MHC class I pathway and CD8+ T cell cross-priming.

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Dendritic Cells Cross-Present Latency Gene Products from Epstein-Barr Virus–Transformed B Cells and Expand Tumor-Reactive Cd8+ Killer T Cells

Journal of Experimental Medicine ◽

10.1084/jem.193.3.405 ◽

2001 ◽

Vol 193 (3) ◽

pp. 405-412 ◽

Cited By ~ 91

Author(s):

Marion Subklewe ◽

Casper Paludan ◽

Ming L. Tsang ◽

Karsten Mahnke ◽

Ralph M. Steinman ◽

...

Keyword(s):

T Cells ◽

Dendritic Cells ◽

Mhc Class I ◽

Cd8 T Cells ◽

Epstein Barr Virus ◽

Class I ◽

Nuclear Antigen ◽

Cross Presentation ◽

Barr Virus ◽

Epstein Barr

Dendritic cells (DCs) are not targets for infection by the transforming Epstein-Barr virus (EBV). To test if the adjuvant role of DCs could be harnessed against EBV latency genes by cross-presentation, DCs were allowed to process either autologous or human histocompatibility leukocyte antigen (HLA)-mismatched, transformed, B lymphocyte cell lines (LCLs) that had been subject to apoptotic or necrotic cell death. After phagocytosis of small numbers of either type of dead LCL, which lacked direct immune-stimulatory capacity, DCs could expand CD8+ T cells capable of killing LCLs that were HLA matched to the DCs. Necrotic EBV-transformed, major histocompatibility complex (MHC) class I–negative LCLs, when presented by DCs, also could elicit responses to MHC class II–negative, EBV-transformed targets that were MHC class I matched to the DCs, confirming efficient cross-presentation of LCL antigens via MHC class I on the DC. Part of this EBV-specific CD8+ T cell response, in both lytic and interferon γ secretion assays, was specific for the EBV nuclear antigen (EBNA)3A and latent membrane protein (LMP)2 latency antigens that are known to be expressed at low levels in transformed cells. The induced CD8+ T cells recognized targets at low doses, 1–10 nM, of peptide. Therefore, the capacity of DCs to cross-present antigens from dead cells extends to the expansion of high affinity T cells specific for viral latency antigens involved in cell transformation.

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A Lipid Based Antigen Delivery System Efficiently Facilitates MHC Class-I Antigen Presentation in Dendritic Cells to Stimulate CD8+ T Cells

Scientific Reports ◽

10.1038/srep27206 ◽

2016 ◽

Vol 6 (1) ◽

Cited By ~ 23

Author(s):

Mithun Maji ◽

Saumyabrata Mazumder ◽

Souparno Bhattacharya ◽

Somsubhra Thakur Choudhury ◽

Abdus Sabur ◽

...

Keyword(s):

T Cells ◽

Dendritic Cells ◽

Antigen Presentation ◽

Mhc Class I ◽

Delivery System ◽

Cd8 T Cells ◽

Class I ◽

Antigen Delivery ◽

Mhc Class I Antigen

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Faculty Opinions recommendation of Dendritic cells cross-dressed with peptide MHC class I complexes prime CD8+ T cells.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1048181.500097 ◽

2006 ◽

Author(s):

Eric Denkers

Keyword(s):

T Cells ◽

Dendritic Cells ◽

Mhc Class I ◽

Cd8 T Cells ◽

Class I

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Faculty Opinions recommendation of Lung CD103+ dendritic cells efficiently transport influenza virus to the lymph node and load viral antigen onto MHC class I for presentation to CD8 T cells.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.13559956.14942056 ◽

2012 ◽

Author(s):

Barney Graham

Keyword(s):

T Cells ◽

Dendritic Cells ◽

Influenza Virus ◽

Lymph Node ◽

Mhc Class I ◽

Cd8 T Cells ◽

Viral Antigen ◽

Class I

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Regulation of Tim-3 function by binding to phosphatidylserine

Biochemical Journal ◽

10.1042/bcj20210652 ◽

2021 ◽

Vol 478 (22) ◽

pp. 3999-4004

Author(s):

Lawrence P. Kane

Keyword(s):

T Cells ◽

Dendritic Cells ◽

T Cell ◽

T Cell Activation ◽

Cd8 T Cells ◽

Cell Activation ◽

Transmembrane Protein ◽

Cytotoxic T Cells ◽

Cross Presentation ◽

First Time

Tim-3 is a transmembrane protein that is highly expressed on subsets of chronically stimulated CD4+ helper and CD8+ cytotoxic T cells, with more transient expression during acute activation and infection. Tim-3 is also constitutively expressed by multiple types of myeloid cells. Like other TIM family members, Tim-3 can bind to phosphatidylserine displayed by apoptotic cells, and this interaction has been shown to mediate uptake of such cells by dendritic cells and cross-presentation of antigens to CD8+ T cells. In contrast, how the recognition of PS by Tim-3 might regulate the function of Tim-3+ T cells is not known. In their recent paper, Lemmon and colleagues demonstrate for the first time that recognition of PS by Tim-3 leads to enhanced T cell activation.

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