Nicholas S. Wilson

Cross-presentation, dendritic cell subsets, and the generation of immunity to cellular antigens

Summary:  Cross‐presentation involves the uptake and processing of exogenous antigens within the major histocompatibility complex (MHC) class I pathway. This process is primarily performed by dendritic cells (DCs), which are not a single cell type but may be divided into several distinct subsets. Those expressing CD8α together with CD205, found primarily in the T‐cell areas of the spleen and lymph nodes, are the major subset responsible for cross‐presenting cellular antigens. This ability is likely to be important for the generation of cytotoxic T‐cell immunity to a variety of antigens, particularly those associated with viral infection, tumorigenesis, and DNA vaccination. At present, it is unclear whether the CD8α‐expressing DC subset captures antigen directly from target cells or obtains it indirectly from intermediary DCs that traffic from peripheral sites. In this review, we examine the molecular basis for cross‐presentation, discuss the role of DC subsets, and examine the contribution of this process to immunity, with some emphasis on DNA vaccination.

Cutting Edge: Generation of Splenic CD8+ and CD8− Dendritic Cell Equivalents in Fms-Like Tyrosine Kinase 3 Ligand Bone Marrow Cultures

We demonstrate that functional and phenotypic equivalents of mouse splenic CD8+ and CD8− conventional dendritic cell (cDC) subsets can be generated in vitro when bone marrow is cultured with fms-like tyrosine kinase 3 (flt3) ligand. In addition to CD45RAhigh plasmacytoid DC, two distinct CD24high and CD11bhigh cDC subsets were present, and these subsets showed equivalent properties to splenic CD8+ and CD8− cDC, respectively, in the following: 1) surface expression of CD11b, CD24, and signal regulatory protein-α; 2) developmental dependence on, and mRNA expression of, IFN regulatory factor-8; 3) mRNA expression of TLRs and chemokine receptors; 4) production of IL-12 p40/70, IFN-α, MIP-1α, and RANTES in response to TLR ligands; 5) expression of cystatin C; and 6) cross-presentation of exogenous Ag to CD8 T cells. Furthermore, despite lacking surface CD8 expression, the CD24high subset contained CD8 mRNA and up-regulated surface expression when transferred into mice. This culture system allows access to bona fide counterparts of the splenic DC subsets.

Systemic activation of dendritic cells by Toll-like receptor ligands or malaria infection impairs cross-presentation and antiviral immunity

The mechanisms responsible for the immunosuppression associated with sepsis or some chronic blood infections remain poorly understood. Here we show that infection with a malaria parasite (Plasmodium berghei) or simple systemic exposure to bacterial or viral Toll-like receptor ligands inhibited cross-priming. Reduced cross-priming was a consequence of downregulation of cross-presentation by activated dendritic cells due to systemic activation that did not otherwise globally inhibit T cell proliferation. Although activated dendritic cells retained their capacity to present viral antigens via the endogenous major histocompatibility complex class I processing pathway, antiviral responses were greatly impaired in mice exposed to Toll-like receptor ligands. This is consistent with a key function for cross-presentation in antiviral immunity and helps explain the immunosuppressive effects of systemic infection. Moreover, inhibition of cross-presentation was overcome by injection of dendritic cells bearing antigen, which provides a new strategy for generating immunity during immunosuppressive blood infections.

Cutting Edge: Conventional CD8α+ Dendritic Cells Are Preferentially Involved in CTL Priming After Footpad Infection with Herpes Simplex Virus-1

CTL play a major role in immunity to HSV type 1, but little is known about the priming process. In this study, we have examined the class I-restricted presentation of an immunodominant determinant from HSV-1 glycoprotein B after footpad infection. We have found that the only cell types capable of presenting this determinant in draining popliteal lymph nodes within the first 3 days after infection are the CD11c+CD8α+CD45RA− dendritic cells. Given that such class I-restricted presentation is essential for CTL priming, this implies that these conventional CD8α+ dendritic cells are the key subset involved in CTL immunity to this virus.

Differential MHC class II synthesis and ubiquitination confers distinct antigen-presenting properties on conventional and plasmacytoid dendritic cells

The importance of conventional dendritic cells (cDCs) in the processing and presentation of antigen is well established, but the contribution of plasmacytoid dendritic cells (pDCs) to these processes, and hence to T cell immunity, remains unclear. Here we showed that unlike cDCs, pDCs continued to synthesize major histocompatibility complex (MHC) class II molecules and the MHC class II ubiquitin ligase MARCH1 long after activation. Sustained MHC class II–peptide complex formation, ubiquitination and turnover rendered pDCs inefficient in the presentation of exogenous antigens but enabled pDCs to continuously present endogenous viral antigens in their activated state. As the antigen-presenting abilities of cDCs and pDCs are fundamentally distinct, these two cell types may activate largely nonoverlapping repertoires of CD4+ T cells.

Regulation of antigen presentation and cross-presentation in the dendritic cell network: facts, hypothesis, and immunological implications.

Dendritic cells (DCs) are central to the maintenance of immunological tolerance and the initiation and control of immunity. The antigen-presenting properties of DCs enable them to present a sample of self and foreign proteins, contained within an organism at any given time, to the T-cell repertoire. DCs achieve this communication with T cells by displaying antigenic peptides bound to MHC I and MHC II molecules. Here we review the studies carried out over the past 15 years to characterize these antigen presentation mechanisms, emphasizing their significance in relation to DC function in vivo. The life cycles of different DC populations found in vivo are described. Furthermore, we provide a critical assessment of the studies that examine the mechanisms controlling DC MHC class II antigen presentation, which have often reached contradictory conclusions. Finally, we review findings pertaining to the biological mechanisms that enable DCs to present exogenous antigens on their MHC class I molecules, a process known as cross-presentation. Throughout, we highlight what we consider to be major knowledge gaps in the field and speculate on possible directions for future research.

Control of MHC class II antigen presentation in dendritic cells: a balance between creative and destructive forces

Summary:  The antigen capturing and presenting abilities of dendritic cells (DCs) are developmentally regulated in a process known as maturation. During maturation, DCs increase several fold their surface expression of major histocompatibility complex class II (MHC II) molecules. This increase is accompanied with a dramatic change in localization of MHC II molecules, which are abundant in endosomal structures in immature DCs but located mostly on the plasma membrane in mature DCs. How these changes relate to antigen processing, generation of MHC II–peptide complexes, and trafficking of MHC II molecules, in the immature and mature states of DC development, has been a matter of debate. Here, we discuss the work carried out to characterize the biochemical and cell biological mechanisms that control MHC II antigen presentation in mouse and human DCs, and how these mechanisms relate to the function of the DC network in vivo. We conclude that the control checkpoints operate downstream of MHC II–peptide complex formation and expression on the plasma membrane, acting in accord with control of MHC II synthesis. Therefore, immature and mature DCs present antigens to T cells under steady state and inflammatory conditions. We advocate that the mechanisms regulating MHC II–peptide complex turnover should be emphasized as an important theme for future DC research.

OX40 engagement depletes intratumoral Tregs via activating FcγRs, leading to antitumor efficacy

Antibodies targeting checkpoint inhibitors or co‐stimulatory receptors on T cells have shown significant antitumor efficacy in preclinical and clinical studies. In mouse tumor models, engagement of activating Fcγ receptor (FcγR)‐expressing immune cells was recently shown to be required for the tumoricidal activity of antibodies recognizing the tumor necrosis factor superfamily receptor (TNFR) GITR (CD357) and CTLA‐4 (CD152). In particular, activating FcγRs facilitated the selective elimination of intratumoral T‐cell populations. However, it remains unclear whether FcγRs contribute to the antitumor efficacy of other immunomodulatory antibodies. Here, we explored the mechanism of antitumor activity mediated by an agonistic antibody (clone OX86) to the co‐stimulatory TNFR OX40 (CD134). OX40 was highly expressed by intratumoral T cells, particularly those of the FoxP3+ regulatory T‐cell (Treg) lineage. OX86 administration resulted in the depletion of intratumoral regulatory T cells in an activating FcγR‐dependent manner, which correlated with tumor regression. Together with previous data from our group and others, these findings support a mechanism whereby antibodies targeting antigens highly expressed by intratumoral T cells can mediate their elimination by FcγR‐expressing immune cells, and facilitate subsequent antitumor immunity.

Normal proportion and expression of maturation markers in migratory dendritic cells in the absence of germs or Toll-like receptor signaling

Dendritic cells (DCs) play major roles in immunosurveillance. In peripheral tissues, ‘immature’ DCs are dedicated to capturing antigens. Detection of pathogens through Toll‐like receptors (TLRs) triggers DC migration to the lymph nodes (LNs), where they acquire a ‘mature’ phenotype specialized at presenting antigens. However, DCs migrate from tissues and mature even in the absence of overt infections. This has been attributed to detection of commensal flora in the skin, the gut or other peripheral tissues in the steady state. To test this assumption, we have analyzed the DCs contained in the lymphoid organs of germ‐free mice and of mice lacking the TLR adapter molecules, MyD88 and TRIF. We show that the proportion and expression of maturation markers in DC immigrants in the LNs of these mice are similar to those in normal mice. These results suggest that DC migration from tissues, followed by their phenotypic maturation, is regulated in the steady state by an inherent program of DC differentiation or by the release of low levels of inflammatory signals from normal tissues.

Lymphoid organ dendritic cells: beyond the Langerhans cells paradigm.

The immune system has developed mechanisms to detect and initiate responses to a continual barrage of immunological challenges. Dendritic cells (DC), a heterogeneous population of leucocytes, play a major role as immunosurveillance agents. To accomplish this function, DC are equipped with highly efficient mechanisms to detect pathogens, to capture, process and present antigens, and to initiate T‐cell responses. These mechanisms are developmentally regulated during the DC life cycle in a process termed ‘maturation’, which was originally defined using Langerhans cells (LC), a DC type of the epidermis. LC exist in the skin in an immature state dedicated to capturing antigens, and in the subcutaneous lymph nodes in a mature state dedicated to presenting those antigens to T cells. The phenotypic changes undergone by LC during maturation, and the correlation of these changes with tissue localization, have been generally considered a paradigm for all DC. However, studies of the multiple DC types found in the lymphoid organs of mice and humans have revealed that most DC subsets do not follow the life cycle typified by LC. In this review we discuss the limitations of the ‘LC paradigm’ and suggest that this model should be revised to accommodate the heterogeneity of the DC system. We also discuss the implications of the maturational status of the DC subsets contained in the lymphoid organs for their putative roles in the induction of immune responses and the maintenance of peripheral tolerance.