Biomedical subjects

Alexis M Kalergis

Publications and source records attributed to Alexis M Kalergis.

13 recordsLinked to original sources

PubMed2007

Injury of skeletal muscle and specific cytokines induce the expression of gap junction channels in mouse dendritic cells.

Dendritic cells (DCs) in culture express at least connexin43, a protein subunit of gap junctions, and form gap junction channels, which could be important for T-cells activation. Here, we evaluated whether DCs express connexins in vivo and also to identify components of their microenvironment that regulate the functional expression of gap junctions. In vivo studies were performed in lymph nodes of mice under control conditions or after skeletal muscle damage. In double immunolabeling studies, connexin45 was frequently detected in DEC205(+) DCs in lymph nodes of control animals, whereas connexin43 was rarely found in DCs. However, connexin43 was upregulated in DCs after skeletal muscle damage. Upregulation of connexin43 gene expression by tissue damage was also confirmed in mice carrying a beta-galactosidase reporter gene in a connexin43 allele. The effect of several cytokines on the expression of functional gap junctions between cultured DCs was also tested. Under control conditions, cultured DCs did not communicate via gap junctions. However, after treatment with keratinocyte-conditioned medium or cytokine mixtures containing at least TNF-alpha and IL-1beta, they became transiently coupled through a pathway sensitive to octanol, a gap junction blocker. Cellular coupling induced by effective cytokine mixtures was prevented by IL-6. Single cytokines (TNF-alpha, IL-1beta, IFN-gamma, or IL-6) or other mixtures than the described above did not induce coupling via gap junctions. Increased levels of connexin43 and connexin45 protein and mRNA accompanied the appearance of cellular coupling. These studies provide demonstration of connexin expression and regulation by specific danger signals in DCs.

Animals↗

PubMed2006

Inhibition of nuclear factor-kappa B enhances the capacity of immature dendritic cells to induce antigen-specific tolerance in experimental autoimmune encephalomyelitis.

Autoimmune disorders develop as a result of deregulated immune responses that target self-antigens and cause destruction of healthy host tissues. Because dendritic cells (DCs) play an important role in the maintenance of peripheral immune tolerance, we are interested in identifying means of enhancing their therapeutic potential in autoimmune diseases. It is thought that during steady state, DCs are able to anergize potentially harmful T cells bearing T cell receptors that recognize self-peptide-major histocompatibility complexes. The tolerogenic capacity of DCs requires an immature phenotype, which is characterized by a reduced expression of costimulatory molecules. On the contrary, activation of antigen-specific naive T cells is enhanced by DC maturation, a process that involves expression of genes controlled by the transcription factor nuclear factor (NF)-kappaB. We evaluated the capacity of drugs that inhibit NF-kappaB to enhance the tolerogenic properties of immature DCs in the experimental autoimmune encephalomyelitis (EAE) model. We show that andrographolide, a bicyclic diterpenoid lactone, and rosiglitazone, a peroxisome proliferator-activated receptor gamma agonist, were able to interfere with NF-kappaB activation in murine DCs. As a result, treated DCs showed impaired maturation and a reduced capacity to activate antigen-specific T cells. Furthermore, NF-kappaB-blocked DCs had an enhanced tolerogenic capacity and were able to prevent EAE development in mice. The tolerogenic feature was specific for myelin antigens and involved the expansion of regulatory T cells. These data suggest that NF-kappaB blockade is a potential pharmacological approach that can be used to enhance the tolerogenic ability of immature DCs to prevent detrimental autoimmune responses.

Animals↗

PubMed2006

Modulation of T cell function by TCR/pMHC binding kinetics.

The interaction between the T cell receptor (TCR) and the peptide-MHC complex (pMHC) at the interface between the T cell and the antigen presenting cell (APC) is the main event controlling the specificity of antigen recognition by T cells. It is thought that TCR/pMHC binding kinetics are critical for the selection of the T cell repertoire in the thymus, as well as the activation of mature T cells in the periphery. One of the binding parameters that conditions T cell activation by pMHC ligands is the half-life of the TCR/pMHC interaction. This kinetic parameter is highly significant for the regulation of T cell activation and therefore determines the capacity of T cells to respond against pathogen- and tumor-derived antigens, avoiding self-reactivity. Several studies support the notion that T cells are activated only by TCR/pMHC interactions that are above a threshold of half-life. pMHC complexes that bind TCRs with half-lives below that threshold behave as null or antagonistic ligands. However, since prolonged half-lives can also impair T cell activation, there seems to be a ceiling for the TCR/pMHC half life that leads to efficient activation of T cells. According to these observations, efficient T cell activation would require an optimal half-life of TCR/pMHC interaction. These kinetic restrictions for T cell activation are important to generate a protective adaptive immune response minimizing cross-reactivity against self-constituents. The nature of the TCR/pMHC interaction defines in the thymus whether a thymocyte develops into a mature T cell or is eliminated by apoptosis. In addition, the kinetics of TCR/pMHC binding can determine the type of response shown by mature T cells in the periphery. Although several studies have focused on the modulation of T cell function by the affinity of the TCR/pMHC interaction, the binding kinetics rules governing T cell activation remain poorly understood. Here we review recent data and propose a new model for the regulation of T cell function by TCR/pMHC binding kinetics.

Animals↗

PubMed2006

Virulent Salmonella enterica serovar typhimurium evades adaptive immunity by preventing dendritic cells from activating T cells.

Dendritic cells (DCs) constitute the link between innate and adaptive immunity by directly recognizing pathogen-associated molecular patterns (PAMPs) in bacteria and by presenting bacterial antigens to T cells. Recognition of PAMPs renders DCs as professional antigen-presenting cells able to prime naïve T cells and initiate adaptive immunity against bacteria. Therefore, interfering with DC function would promote bacterial survival and dissemination. Understanding the molecular mechanisms that have evolved in virulent bacteria to evade activation of adaptive immunity requires the characterization of virulence factors that interfere with DC function. Salmonella enterica serovar Typhimurium, the causative agent of typhoid-like disease in the mouse, can prevent antigen presentation to T cells by avoiding lysosomal degradation in DCs. Here, we show that this feature of virulent Salmonella applies in vivo to prevent activation of adaptive immunity. In addition, this attribute of virulent Salmonella requires functional expression of a type three secretion system (TTSS) and effector proteins encoded within the Salmonella pathogenicity island 2 (SPI-2). In contrast to wild-type virulent Salmonella, mutant strains carrying specific deletions of SPI-2 genes encoding TTSS components or effectors proteins are targeted to lysosomes and are no longer able to prevent DCs from activating T cells in vitro or in vivo. SPI-2 mutant strains are attenuated in vivo, showing reduced tissue colonization and enhanced T-cell activation, which confers protection against a challenge with wild-type virulent Salmonella. Our data suggest that impairment of DC function by the activity of SPI-2 gene products is crucial for Salmonella pathogenesis.

Animals↗

PubMed2006

The dendritic cell-T cell synapse as a determinant of autoimmune pathogenesis.

Autoimmune diseases occur when the immune response is targeted to self-antigens, leading to destruction or altered function of specific cells and tissues. Although the aetiology of these diseases has not yet been fully elucidated, it is believed that genetically determined susceptibility and environmental triggers are both implicated in the detrimental immune response against the body's own tissues. Dendritic cells (DCs) are professional antigen presenting cells that play an important role in maintaining peripheral tolerance by preventing self-reactive T cells from causing autoimmune damage. Thus, alterations in the physiology of DCs are likely to be responsible for defective immune regulatory mechanisms and incomplete tolerance to self. Here, we will focus specifically on the ways in which the immunological synapse occurring at the DC-T cell interface can fine-tune the balance between tolerance and immunity and how alterations of this synapse can determine induction or perpetuation of autoimmune responses. Activating/inhibitory receptors expressed on the surface of DCs and T cells modulate the function of these cells and influence the course of the immune response. Pharmacological approaches that can modulate DC function will be also addressed as a potential antigen-specific strategy in the design of new, noninvasive therapies to prevent or to treat chronic inflammatory autoimmune disorders.

Animals↗

PubMed2005

T cell receptor binding kinetics required for T cell activation depend on the density of cognate ligand on the antigen-presenting cell.

CD8(+) T cells recognize peptides of eight to nine amino acid residues long in the context of MHC class I molecules on the surface of antigen-presenting cells (APCs). This recognition event is highly sensitive, as evidenced by the fact that T cells can be activated by cognate peptide/MHC complex (pMHC) at extremely low densities (1-50 molecules). High sensitivity is particularly valuable for detection of antigens at low density, such as those derived from tumor cells and intracellular pathogens, which can down-modulate cognate pMHCs from the surface of APCs to evade recognition by the adaptive immune system. T cell activation is only triggered in response to interactions between the T cell receptor (TCR) and the pMHC ligand that reach a specific half-life threshold. However, interactions with excessively long half-lives result in impaired T cell activation. Thus, efficient T cell activation by pMHC on the surface of APCs requires an optimal dwell time of TCR-pMHC interaction. Here, we show that, although this is a requirement at low cognate pMHC density on the APC surface, at high epitope density there is no impairment of T cell activation by extended TCR-pMHC dwell times. This observation was predicted by mathematical simulations for T cell activation by pMHC at different densities and supported by experiments performed on APCs selected for varied expression of cognate pMHC. According to these results, effective T cell activation depends on a complex interplay between inherent TCR-pMHC binding kinetics and the epitope density on the APC.

Antigen-Presenting Cells↗

PubMed2005

Molecular interactions between dendritic cells and Salmonella: escape from adaptive immunity and implications on pathogenesis.

Dendritic cells (DCs) constitute the link between innate and adaptive immunity by directly recognizing pathogen-associated molecular patterns (PAMPs) on bacteria and by processing and presenting bacterial antigens to T cells. Recognition of PAMPs renders DCs as professional antigen-presenting cells with the ability to prime naive T cells and to initiate the adaptive immune response against pathogen-derived antigens. For this reason, any interference with DC function might be advantageous for bacterial survival and dissemination. Identification of the molecular interactions occurring between DCs and bacterial pathogens is necessary to understand the mechanisms that virulent bacteria have evolved to prevent recognition by the adaptive immune system. This could be helpful in the identification of possible new targets that might lead to the design of effective therapies aimed at preventing or treating serious infections by these pathogens. In this article, we focus on Salmonella enterica serovar Typhimurium, the causative agent of typhoid-like disease in the mouse, and how it is able to escape from DC-mediated antigen presentation by avoiding lysosomal degradation. This feature of virulent Salmonella requires the functional expression of the Type Three Secretion System (TTSS) and effector proteins encoded within the Salmonella pathogenicity island 2 (SPI-2). Recent studies have demonstrated that impairment of DC function by the activity of SPI-2 gene products is crucial for Salmonella pathogenesis.

Animals↗

PubMed2004

Salmonella escape from antigen presentation can be overcome by targeting bacteria to Fc gamma receptors on dendritic cells.

Dendritic cells (DCs) are professional APCs with the unique ability to activate naive T cells, which is required for initiation of the adaptive immune response against pathogens. Therefore, interfering with DC function would be advantageous for pathogen survival and dissemination. In this study we provide evidence suggesting that Salmonella enterica serovar typhimurium, the causative agent of typhoid disease in the mouse, interferes with DC function. Our results indicate that by avoiding lysosomal degradation, S. typhimurium impairs the ability of DCs to present bacterial Ags on MHC class I and II molecules to T cells. This process could correspond to a novel mechanism developed by this pathogen to evade adaptive immunity. In contrast, when S. typhimurium is targeted to FcgammaRs on DCs by coating bacteria with Salmonella-specific IgG, bacterial Ags are efficiently processed and presented on MHC class I and class II molecules. This enhanced Ag presentation leads to a robust activation of bacteria-specific T cells. Laser confocal microscopy experiments show that virulent S. typhimurium is rerouted to the lysosomal degradation pathway of DCs when internalized through FcgammaR. These observations are supported by electron microscopy studies demonstrating that internalized S. typhimurium shows degradation signs only when coated with IgG and captured by FcgammaRs on DCs. Therefore, our data support a potential role for bacteria-specific IgG on the augmentation of Ag processing and presentation by DCs to T cells during the immune response against intracellular bacteria.

Animals↗

PubMed2004

Andrographolide interferes with T cell activation and reduces experimental autoimmune encephalomyelitis in the mouse.

Andrographolide is a bicyclic diterpenoid lactone derived from extracts of Andrographis paniculata, a plant indigenous to South Asian countries that shows anti-inflammatory properties. The molecular and cellular bases for this immunomodulatory capacity remain unknown. Here, we show that andrographolide is able to down-modulate both humoral and cellular adaptive immune responses. In vitro, this molecule was able to interfere with T cell proliferation and cytokine release in response to allogenic stimulation. These results were consistent with the observation that T cell activation by dendritic cells (DCs) was completely abolished by exposing DCs to andrographolide during antigen pulse. This molecule was able to interfere with maturation of DCs and with their ability to present antigens to T cells. Furthermore, in vivo immune responses such as antibody response to a thymus-dependent antigen and delayed-type hypersensitivity were drastically diminished in mice by andrographolide treatment. Finally, the ability of andrographolide to inhibit T cell activation was applied to interfere with the onset of experimental autoimmune encephalomyelitis (EAE), an inflammatory demyelinating disease of the central nervous system that is primarily mediated by CD4(+) T cells and serves as an animal model for human multiple sclerosis. Treatment with andrographolide was able to significantly reduce EAE symptoms in mice by inhibiting T cell and antibody responses directed to myelin antigens. Our data suggest that andrographolide is able to efficiently block T cell activation in vitro, as well as in vivo, a feature that could be useful for interfering with detrimental T cell responses.

Animals↗

PubMed2004

[Clinical applications of antigen-specific detection of T lymphocytes].

The important role that T lymphocytes play during the immune response against pathogens and tumor cells has encouraged the development of technologies aimed to detect these immune cells in an antigen-specific fashion. Recently, a methodology consisting of the production of soluble ligands that bind the T lymphocyte receptor was developed. Such ligands consist of molecules from the Major Histocompatibility Complex loaded with either pathogen or tumor derived peptides. These molecular complexes are tetramerized to enhance the binding efficiency to the T cell receptor (TCR), which improves the detection sensitivity of antigen specific T lymphocytes. This new technology is currently used successfully during the follow up patients that were vaccinated against certain tumor antigens, to evaluate the expansion of tumor specific T lymphocytes. This methodology is also promising for the detection and specific deletion of autoreactive T lymphocytes as a treatment for autoimmune disorders. In this article we review the molecular components involved in antigen recognition by T lymphocytes, and the potential benefits for Biomedicine that could result from the detection of these cells by soluble specific ligands.

Humans↗

PubMed2003

Modulation of T cell immunity by TCR/pMHC dwell time and activating/inhibitory receptor pairs on the antigen-presenting cell.

The molecular interactions occurring at the interface between the antigen presenting cell (APC) and the T lymphocyte play an important role in the immune surveillance against infectious agents and tumors, as well as in autoimmunity and transplant rejection. The significance of the APC-T cell interaction in immunity is underscored by the observation that deficiencies in the function of either one of these two cell types cause extreme susceptibility to infections and tumor growth. Furthermore, a disregulated APC-T cell interaction can initiate autoimmunity. Thus, antigen recognition by T cells must be tightly regulated in order to ensure protection against pathogens and tumors, avoiding activation of self-reactive T cells. Efficient T cell activation requires two simultaneous signals provided by the APC: Antigen (or signal 1) and co-stimulation (or signal 2). The specificity of antigen recognition by T cells (signal 1) is controlled exclusively by the T cell receptor (TCR), an extremely diverse heterodimeric protein composed of disulfide-bonded alpha and beta chains. While it is clear that the TCR recognizes antigens as small peptides bound to molecules of the Major Histocompatibility Complex (MHC), the molecular explanation for the specificity of antigen recognition by the betaalphaTCR is just beginning to be elucidated. In this review are described some of the advances made in the understanding of the molecular interactions that define the antigen-specificity of the TCR, and the current models for T cell activation by antigen on APCs are discussed.

Animals↗

PubMed2002

Activated TCRs remain marked for internalization after dissociation from pMHC.

To assess the roles of serial engagement and kinetic proofreading in T cell receptor (TCR) internalization, we have developed a mathematical model of this process. Our determination of TCR down-regulation for an array of TCR mutants, interpreted in the context of the model, has provided new information about peptide-induced TCR internalization. The amount of TCR down-regulation increases to a maximum value and then declines as a function of the half-life of the bond between the TCR and peptide-major histocompatibility complex (pMHC). The model shows that this behavior, which reflects competition between serial engagement and kinetic proofreading, arises only if it is postulated that activated TCRs remain marked for internalization after dissociation from pMHC. The model also predicts that because of kinetic proofreading, the range of TCR-pMHC-binding half-lives required for T cell activation depends on the concentrations and localization of intracellular signaling molecules. We show here that kinetic proofreading provides an explanation for the different requirements for activation observed in naïve and memory T cells.

Animals↗

PubMed2002

Inducing tumor immunity through the selective engagement of activating Fcgamma receptors on dendritic cells.

Induction of tumor-specific immunity requires that dendritic cells (DCs) efficiently capture and present tumor antigens to result in the expansion and activation of tumor-specific cytotoxic T cells. The transition from antigen capture to T cell stimulation requires a maturation signal; in its absence tolerance, rather than immunity may develop. While immune complexes (ICs) are able to enhance antigen capture, they can be poor at inducing DC maturation, naive T cell activation and protective immunity. We now demonstrate that interfering with the inhibitory signal delivered by FcgammaRIIB on DCs converts ICs to potent maturation agents and results in T cell activation. Applying this approach to immunization with DCs pulsed ex-vivo with ICs, we have generated antigen-specific CD8+ T cells in vivo and achieved efficient protective immunity in a murine melanoma model. These data imply that ICs may normally function to maintain tolerance through the binding to inhibitory FcgammaRs on DCs, but they can be converted to potent immunogenic stimuli by selective engagement of activating FcgammaRs. This mechanism suggests a novel approach to the development of tumor vaccines.

Animals↗