Cell Biology Of Antigen Presentation Research Articles

Borrelia burgdorferi-Induced Changes in the Class II Self-Immunopeptidome Displayed on HLA-DR Molecules Expressed by Dendritic Cells.

The MHC class II antigen processing and presentation pathway has evolved to derive short amino acid peptides from proteins that enter the endocytic pathway, load them onto MHC class II molecules and display them on the surface of antigen presenting cells for recognition by CD4+ T cells. Under normal circumstances, peptides bound to MHC class II molecules are derived from host (self) proteins and not recognized by T cells due to tolerance mechanisms. Pathogens induce significant changes in the biology of antigen presenting cells, including upregulation of MHC processing and presentation. We therefore hypothesized that exposure to pathogens may alter the repertoire of self-peptides bound to MHC class II molecules. To test this hypothesis, we isolated monocyte-derived dendritic cells from healthy subjects, exposed them to the TLR-2 agonist lipoteichoic acid or live Borrelia burgdorferi, the causative agent of Lyme disease, and isolated and characterized HLA-DR associated peptides using mass spectrometry. Our results show that lipoteichoic acid-stimulated, B. burgdorferi-stimulated and unstimulated monocyte-derived dendritic cells largely derive their self-peptides from similar overlapping sets of host proteins. However, lipoteichoic acid and B. burgdorferi stimulation promote the processing and presentation of new sets of HLA-DR associated self-peptides derived from unique protein sources. Examination of processes and compartments these proteins reside in, indicate that activation of monocyte-derived dendritic cells changes the range of host self-proteins available for processing and presentation on MHC class II molecules. These findings reveal that the HLA-DR-bound self-immunopeptidome presented by mo-DCs is dynamic in nature and changes with activation state reflective of cellular function. In addition, among the repertoire of self-peptides bound to HLA-DR are several epitopes known to be recognized by autoreactive T cells. These studies are relevant to our basic understanding of pathogen-induced changes in monocyte-derived dendritic cell function, and the mechanisms involved in infection-induced autoimmune illnesses such as Lyme arthritis.

Understanding the Role of the Unfolded Protein Response Sensor IRE1 in the Biology of Antigen Presenting Cells.

The unfolded protein response (UPR) is an adaptive response that maintains the fidelity of the cellular proteome in conditions that subvert the folding capacity of the cell, such as those noticed in infection and inflammatory contexts. In immunity, the UPR sensor IRE1 (Inositol-requiring enzyme 1-alpha) has emerged as a critical regulator of the homeostasis of antigen presenting cells (APCs). In the past few years, it has become clear that IRE1 plays canonical and non-canonical roles in APCs, many of which intersect with key features of these cells, including the initiation of inflammation, antibody production, and antigen presentation. The aims of the present review are to provide recent insights on the mechanisms by which IRE1 regulates the diversity of APC functions and to highlight its relevance in the coordination of innate and adaptive immunity.

Imaging Flow Cytometry to Assess Antigen-Presenting-Cell Function.

This unit describes methods for quantifying phagocytosis and imaging the immunological synapse between T cells and antigen-presenting cells (APCs), with both techniques delivering valuable information about APC function. These aspects of APC biology have traditionally been challenging to quantify, and imaging flow cytometry, which harnesses the high-throughput nature of flow cytometry combined with the capacity of microscopy to deliver spatial localization, facilitates analysis of these APC functions in a fashion that was previously not possible. Imaging flow cytometry allows large numbers of events to be captured and large amounts of fluorescence data to be quantified at the physical location of markers of interest, both on the cell surface and in intracellular compartments, combining key features of traditional flow cytometry and fluorescence microscopy. © 2019 by John Wiley & Sons, Inc.

Tetraspanins as Organizers of Antigen-Presenting Cell Function.

Professional antigen-presenting cells (APCs) include dendritic cells, monocytes, and B cells. APCs internalize and process antigens, producing immunogenic peptides that enable antigen presentation to T lymphocytes, which provide the signals that trigger T-cell activation, proliferation, and differentiation, and lead to adaptive immune responses. After detection of microbial antigens through pattern recognition receptors (PRRs), APCs migrate to secondary lymphoid organs where antigen presentation to T lymphocytes takes place. Tetraspanins are membrane proteins that organize specialized membrane platforms, called tetraspanin-enriched microdomains, which integrate membrane receptors, like PRR and major histocompatibility complex class II (MHC-II), adhesion proteins, and signaling molecules. Importantly, through the modulation of the function of their associated membrane partners, tetraspanins regulate different steps of the immune response. Several tetraspanins can positively or negatively regulate the activation threshold of immune receptors. They also play a role during migration of APCs by controlling the surface levels and spatial arrangement of adhesion molecules and their subsequent intracellular signaling. Finally, tetraspanins participate in antigen processing and are important for priming of naïve T cells through the control of T-cell co-stimulation and MHC-II-dependent antigen presentation. In this review, we discuss the role of tetraspanins in APC biology and their involvement in effective immune responses.

A major isoform of the E3 ubiquitin ligase March-I in antigen-presenting cells has regulatory sequences within its gene

Regulation of major histocompatibility complex class II (MHC-II) expression is important not only to maintain a diverse pool of MHC-II-peptide complexes but also to prevent development of autoimmunity. The membrane-associated RING-CH (March) E3 ubiquitin ligase March-I regulates ubiquitination and turnover of MHC-II-peptide complexes in resting dendritic cells (DCs) and B cells. However, activation of either cell type terminates March-I expression, thereby stabilizing MHC-II-peptide complexes. Despite March-I's important role in the biology of antigen-presenting cells (APCs), how expression of March-I mRNA is regulated remains unknown. We now show that both DCs and B cells possess a distinct isoform of March-I whose expression is regulated by a promoter located within the March-I gene. Using March-I promoter fragments to drive expression of GFP, we also identified a core promoter for expression of March-I in DCs and B cells, but not in fibroblasts, kidney cells, or epithelial cells, that contains regulatory regions that down-regulate March-I expression upon activation of DCs. Curiously, we found downstream sequence elements, present in the first coding exon of March-I in APCs, that confer regulation of March-I expression in activated APCs. In summary, our study identifies regulatory regions of the March-I gene that confer APC-specific expression and activation-induced modulation of March-I expression in DCs and B cells.

ERAP1-ERAP2 dimers trim MHC I-bound precursor peptides; implications for understanding peptide editing.

The processing of MHC class I antigenic precursor peptides by the endoplasmic reticulum aminopeptidase 1 (ERAP1) and ERAP2 is an important event in the cell biology of antigen presentation. To date, the molecular context by which the ERAP enzymes trim precursor peptides, and how ERAPs shape peptide repertoires, remain open questions. Using ERAP1 and ERAP2 heterodimers (ERAP1/2), and N-terminally extended model and natural peptides in their free and HLA-B*0801-bound forms, we characterized the mode of action of ERAPs. We provide evidence that ERAP1/2 can trim MHC I-bound precursor peptides to their correct and final lengths, albeit more slowly than the corresponding free precursors. Trimming of MHC I-bound precursors by ERAP1/2 increases the conformational stability of MHC I/peptide complexes. From the data, we propose a molecular mechanistic model of ERAP1/2 as peptide editors. Overall, our study provides new findings on a significant issue of the ERAP-mediated processing pathway of MHC class I antigens.

Modulation of MHC class I antigen presentation by the adenovirus E3-19K protein (100.8)

Abstract The presentation of viral antigens by MHC I to CTLs is vital for elimination of infected cells. In turn, viruses have evolved various strategies to abrogate antiviral cellular immune responses. For example, the adenovirus (Ad) E3-19K protein retains MHC I in the endoplasmic reticulum, dramatically suppressing the cell-surface presentation of viral antigens. Many aspects of how E3-19K exerts its function towards MHC I are poorly understood. This knowledge is important as the E3-19K/MHC I interaction is thought to play a role in enabling Ads to cause persistent infections. We characterized biophysically and cellularly interaction between E3-19K proteins of different Ad serotypes (Ad 7 and 35, subgroup B; Ad 2 and Ad 5, subgroup C; Ad 37, subgroup D; and Ad 4, subgroup E) and HLA-A, -B, and -C molecules. Our results show that E3-19K proteins from Ad serotypes of the same subgroup display more similar binding properties for a given MHC I than those of different subgroups. Furthermore, E3-19K proteins exhibited allele- and locus-specificity; higher avidity for HLA-A relative to -B molecules, and no interaction with HLA-C molecules. FACS analysis showed that binding affinities correlated (in a negative way) with levels of MHC I expression on infected cells. Finally, we propose the first model of interaction between E3-19K and MHC I. Overall, our studies provide novel insights into the structure/function relationship of E3-19K and the molecular cell biology of antigen presentation.

Inhibition of mouse TAP by immune evasion molecules encoded by non-murine herpesviruses

Herpesviruses escape elimination by cytotoxic T lymphocytes through specific interference with the antigen-presenting function of MHC class I (MHC I) molecules. The transporter associated with antigen processing (TAP) forms a bottleneck in the MHC I antigen presentation pathway. The fact that multiple viruses, especially herpesviruses, encode molecules blocking TAP function is a case in point. The action of these viral immuno evasins is usually potent and very specific, making these proteins valuable tools for studying the cell biology of antigen presentation, including alternative antigen processing pathways. Yet, no dedicated TAP inhibitor has been described for any of the mouse herpesviruses.To permit the use of immuno evasins derived from non-mouse herpesviruses in mouse models, we assessed the cross-species activity of four TAP inhibitors and one tapasin inhibitor in the context of three different mouse haplotypes, H-2b, H-2d, and H-2k. Two of the four TAP inhibitors, the bovine herpesvirus 1-encoded UL49.5 and the human cytomegalovirus (HCMV)-encoded US6 protein, potently inhibited mouse TAP. ICP47 and BNLF2a, encoded by herpes simplexvirus 1 and Epstein–Barr virus, respectively, failed to inhibit TAP in all mouse cells tested. Previous work, however, demonstrated that US6 did not cross the mouse species barrier. We now show that substitution of the cysteine residue at position 108 was responsible for this lack of activity. The HCMV-encoded tapasin inhibitor US3 efficiently downregulated H-2d molecules on 3T3 cells, but not in other cell lines tested. Finally, we show that synthetic peptides comprising the functional domain of US6 can be exploited as a versatile TAP inhibitor. In conclusion, a complete overview is presented of the applicability of herpesvirus-encoded TAP and tapasin inhibitors in mouse cells of different genetic background.

Grand Challenges in Molecular Antigen-presenting Cell Biology

SPECIALTY GRAND CHALLENGE article Front. Immunol., 21 March 2011Sec. Antigen Presenting Cell Biology volume 2 - 2011 | https://doi.org/10.3389/fimmu.2011.00008

Toward a Molecular Explanation for Cross-presentation of Antigens to the Immune System

Cross-presentation is increasingly recognized as a key mechanism by which specific antigen-presenting cells (APCs) activate the cellular immune system against virally infected or neoplastic cells. These APCs have the capacity to acquire exogenous proteins by phagocytosis or endocytosis, derive antigenic peptides, and then cross-present them in the context of class I major histocompatibility complex molecules. Because APCs provide both an antigenic stimulus and costimulatory signals, they can effectively activate naive CD8+ T lymphocytes, resulting in a brisk cellular immune response. The precise cellular pathways that permit an exogenous antigen to be presented in the context of class I major histocompatibility complex is the focus of intense investigation that has illuminated our understanding of the cell biology of APCs. This article reviews our understanding of how APCs cross-present antigen, illustrating how this process differs from the canonical pathways of antigen presentation. We define the central players required for cross-presentation and then focus on recent studies that reveal the molecular mechanisms underlying this phenomenon. Understanding these mechanisms will likely inform the development of effective cell-based vaccines and other cellular immunotherapies and is therefore of interest to practitioners of transfusion medicine.

The Role of Antigen-presenting Cells in HIV Pathogenesis.

The study of antigen-presenting cells (APC) in HIV pathogenesis has been ongoing for almost 20 years. The initial studies recognized the important role of APC as targets for HIV infection and their ability to serve as reservoirs of virus, particularly in tissues. The issue of whether HIV impacts the functional competency of APC has been more controversial, with some studies showing reduced expression of important costimulatory molecules on APC, but others showing the functional capacity of APC to be normal. The study of APC has advanced with recent interest in one class of APC, namely the dendritic cell. These cells have been shown to consist of numerous subsets and serve an important role in bridging innate and adaptive immune responses. The impact of HIV infection on dendritic cells has recently been characterized, as well as the critical functional role of these cells in host defenses in HIV-infected patients. One of the more exciting recent advances in APC biology is the ability to manipulate APC ex vivo for therapeutic purposes in an attempt to restore immune responses in HIV-infected persons. This review covers many of the advances of the field of APC biology and puts them into perspective with HIV pathogenesis.

The Cell Biology of Antigen Presentation

The Cell Biology of Antigen Presentation

Antigen-presenting cells in allergy

The complex interaction of the innate and adaptive immune system requires flexibility and cooperation among various cell types. In this regard, antigen-presenting-cells (APCs) play a pivotal role in transferring information from the periphery of the organism to lymphoid organs, where they initiate the activation of naive T cells. Dendritic cells, Langerhans’ cells (LCs), and macrophages are also critical in the induction of allergic inflammation by presenting allergens to T lymphocytes and by contributing to the local recruitment of effector cells. Because of a complex genetic background, atopic individuals exhibit a dysregulation of T cell–mediated immune mechanisms. Attempts to understand the role APCs play in these pathophysiologic conditions are in progress and may allow development of new treatment strategies. In this review we will focus on the biology of APCs and their unique role in the induction and control of allergic inflammation. (J Allergy Clin Immunol 2001;108:329-39.)

Inhibition of MHC class I antigen presentation by viral proteins.

An essential part of the immune response to viral infections is the recognition and elimination of infected cells by cytotoxic T lymphocytes. For this purpose a display mechanism has evolved which is present in almost all nucleated cells: the major histocompatibility complex class I antigen processing pathway. Both self and foreign antigens are degraded in the cytosol to peptides which are translocated into the endoplasmic reticulum where they are loaded onto MHC class I molecules. Pathogens living inside the cell are evolving under the constant selection pressure of such immune surveillance. As a result such infectious organisms have developed a variety of strategies to prevent their antigens from being presented. Since our understanding of the cell biology of antigen presentation has greatly advanced in recent years, it has now become possible to unravel several of the molecular mechanisms by which viruses interfere with MHC class I antigen presentation. Examples for the interference of viral molecules with components of the MHC class I pathway are presented in this review.

The biochemistry and cell biology of antigen presentation by MHC class I and class II molecules. Implications for development of combination vaccines.

T lymphocytes play a central role in adaptive immunity. They provide direct effector function, regulate the activity of non-antigen-specific effector cells such as macrophages, and control the production of antibodies by B cells. Thus, the proper stimulation of T cells is critical to effective vaccination. T cells bearing alpha beta receptors are stimulated by antigen-derived peptides displayed on cell surfaces bound to highly polymorphic, major histocompatibility complex-encoded glycoproteins. To elicit suitable T cell responses vaccines must, therefore, contain proteins or peptides derived from the organism against which protection is desired, the pathogen-derived peptides must be capable of interacting with the allelic forms of the MHC molecules expressed in the vaccinated individuals, and the vaccine components must be delivered in a manner that ensures they are made available for binding to the MHC molecules on appropriate antigen-presenting cells. This paper has reviewed the rules governing peptide binding to MHC molecules, the intracellular pathways of protein synthesis, protein degradation, and protein and peptide transport involved in bringing together antigenic peptides and MHC molecules, and the distinct function of MHC class I versus class II molecules. The implications of this knowledge for effective combined vaccine design and delivery were considered.

Dendritic Cells as Antigen Presenting Cellsin Vivo

The biology of antigen presenting cells (APC) traditionally is studied in tissue culture systems using T cells that have been expanded beforehand by stimulation with antigen. Here we consider the distinctive roles of dendritic cells for sensitizing or priming T cells both in vitro and in vivo. Several functions of dendritic cells have been identified in tissue culture that are pertinent to T cell sensitization. These include the ability to a) capture and retain foreign antigens in an immunogenic form, b) bind antigen-specific resting lymphocytes, and c) activate T cells to produce lymphokines and undergo long term clonal growth. Dendritic cells have several properties in vivo that also would contribute to APC function. These are a) their widespread tissue distribution permitting access to antigens in most organs, b) the capacity to home via the blood stream and afferent lymph to the T-dependent areas of spleen and lymph node, and c) the ability to capture antigen in antigen-pulsed animals. Dendritic cells bearing antigen have been administered in situ to initiate responses like contact sensitivity, graft rejection, and antibody formation. A most striking recent example is that, when dendritic cells are pulsed with protein antigens in vitro and administered to immunologically naive mice, there is direct priming of antigen-specific T cells that are restricted to the MHC of the injected APC.