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1.
PLoS Pathog ; 11(6): e1004941, 2015 Jun.
Article in English | MEDLINE | ID: mdl-26107264

ABSTRACT

Naïve anti-viral CD8+ T cells (TCD8+) are activated by the presence of peptide-MHC Class I complexes (pMHC-I) on the surface of professional antigen presenting cells (pAPC). Increasing the number of pMHC-I in vivo can increase the number of responding TCD8+. Antigen can be presented directly or indirectly (cross presentation) from virus-infected and uninfected cells, respectively. Here we determined the relative importance of these two antigen presenting pathways in mousepox, a natural disease of the mouse caused by the poxvirus, ectromelia (ECTV). We demonstrated that ECTV infected several pAPC types (macrophages, B cells, and dendritic cells (DC), including DC subsets), which directly presented pMHC-I to naïve TCD8+ with similar efficiencies in vitro. We also provided evidence that these same cell-types presented antigen in vivo, as they form contacts with antigen-specific TCD8+. Importantly, the number of pMHC-I on infected pAPC (direct presentation) vastly outnumbered those on uninfected cells (cross presentation), where presentation only occurred in a specialized subset of DC. In addition, prior maturation of DC failed to enhance antigen presentation, but markedly inhibited ECTV infection of DC. These results suggest that direct antigen presentation is the dominant pathway in mice during mousepox. In a broader context, these findings indicate that if a virus infects a pAPC then the presentation by that cell is likely to dominate over cross presentation as the most effective mode of generating large quantities of pMHC-I is on the surface of pAPC that endogenously express antigens. Recent trends in vaccine design have focused upon the introduction of exogenous antigens into the MHC Class I processing pathway (cross presentation) in specific pAPC populations. However, use of a pantropic viral vector that targets pAPC to express antigen endogenously likely represents a more effective vaccine strategy than the targeting of exogenous antigen to a limiting pAPC subpopulation.


Subject(s)
Antigen Presentation/immunology , CD8-Positive T-Lymphocytes/immunology , Histocompatibility Antigens Class I/immunology , Lymphocyte Activation/immunology , Animals , Cross-Priming/immunology , Dendritic Cells/immunology , Macrophages/immunology , Mice, Inbred C57BL , Mice, Transgenic , Peptides/immunology , Peptides/metabolism , Phenotype
2.
J Immunol ; 186(1): 183-94, 2011 Jan 01.
Article in English | MEDLINE | ID: mdl-21098225

ABSTRACT

Protracted psychological stress elevates circulating glucocorticoids, which can suppress CD8(+) T cell-mediated immunity, but the mechanisms are incompletely understood. Dendritic cells (DCs), required for initiating CTL responses, are vulnerable to stress/corticosterone, which can contribute to diminished CTL responses. Cross-priming of CD8(+) T cells by DCs is required for initiating CTL responses against many intracellular pathogens that do not infect DCs. We examined the effects of stress/corticosterone on MHC class I (MHC I) cross-presentation and priming and show that stress/corticosterone-exposed DCs have a reduced ability to cross-present OVA and activate MHC I-OVA(257-264)-specific T cells. Using a murine model of psychological stress and OVA-loaded ß(2)-microglobulin knockout "donor" cells that cannot present Ag, DCs from stressed mice induced markedly less Ag-specific CTL proliferation in a glucocorticoid receptor-dependent manner, and endogenous in vivo T cell cytolytic activity generated by cross-presented Ag was greatly diminished. These deficits in cross-presentation/priming were not due to altered Ag donation, Ag uptake (phagocytosis, receptor-mediated endocytosis, or fluid-phase uptake), or costimulatory molecule expression by DCs. However, proteasome activity in corticosterone-treated DCs or splenic DCs from stressed mice was partially suppressed, which limits formation of antigenic peptide-MHC I complexes. In addition, the lymphoid tissue-resident CD11b(-)CD24(+)CD8α(+) DC subset, which carries out cross-presentation/priming, was preferentially depleted in stressed mice. At the same time, CD11b(-)CD24(+)CD8α(-) DC precursors were increased, suggesting a block in development of CD8α(+) DCs. Therefore, glucocorticoid-induced changes in both the cellular composition of the immune system and intracellular protein degradation contribute to impaired CTL priming in stressed mice.


Subject(s)
Corticosterone/physiology , Cross-Priming/immunology , Cytotoxicity, Immunologic/drug effects , Dendritic Cells/immunology , Immunosuppression Therapy , Lymphocyte Activation/drug effects , Stress, Psychological/immunology , T-Lymphocytes, Cytotoxic/immunology , Animals , Cells, Cultured , Coculture Techniques , Corticosterone/biosynthesis , Cross-Priming/drug effects , Cytotoxicity, Immunologic/immunology , Dendritic Cells/metabolism , Immobilization , Lymphocyte Activation/immunology , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Mice, Transgenic , Ovalbumin/immunology , Ovalbumin/metabolism , T-Lymphocytes, Cytotoxic/cytology , T-Lymphocytes, Cytotoxic/metabolism
3.
J Immunol ; 184(4): 1867-75, 2010 Feb 15.
Article in English | MEDLINE | ID: mdl-20089700

ABSTRACT

The systemic elevation of psychological stress-induced glucocorticoids strongly suppresses CD8(+) T cell immune responses resulting in diminished antiviral immunity. However, the specific cellular targets of stress/glucocorticoids, the timing of exposure, the chronology of immunological events, and the underlying mechanisms of this impairment are incompletely understood. In this study, we address each of these questions in the context of a murine cutaneous HSV infection. We show that exposure to stress or corticosterone in only the earliest stages of an HSV-1 infection is sufficient to suppress, in a glucocorticoid receptor-dependent manner, the subsequent antiviral immune response after stress/corticosterone has been terminated. This suppression resulted in early onset and delayed resolution of herpetic lesions, reduced viral clearance at the site of infection and draining popliteal lymph nodes (PLNs), and impaired functions of HSV-specific CD8(+) T cells in PLNs, including granzyme B and IFN-gamma production and the ability to degranulate. In knockout mice lacking glucocorticoid receptors only in T cells, we show that these impaired CD8(+) T cell functions are not due to direct effects of stress/corticosterone on the T cells, but the ability of PLN-derived dendritic cells to prime HSV-1-specific CD8(+) T cells is functionally impaired. These findings highlight the susceptibility of critical early events in the generation of an antiviral immune response to neuroendocrine modulation and implicate dendritic cells as targets of stress/glucocorticoids in vivo. These findings also provide insight into the mechanisms by which the clinical use of glucocorticoids contributes to altered immune responses in patients with viral infections or tumors.


Subject(s)
Corticosterone/pharmacology , Dendritic Cells/immunology , Dendritic Cells/pathology , Herpes Simplex/immunology , Herpes Simplex/pathology , Herpesvirus 1, Human/immunology , Immunosuppressive Agents/pharmacology , Stress, Psychological/immunology , Animals , CD8-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/pathology , CD8-Positive T-Lymphocytes/virology , Corticosterone/metabolism , Dendritic Cells/virology , Herpes Simplex/virology , Immobilization , Immunosuppressive Agents/metabolism , Lymph Nodes/immunology , Lymph Nodes/pathology , Lymph Nodes/virology , Male , Mice , Mice, Inbred C57BL , Mice, Transgenic , Viral Envelope Proteins/immunology
4.
Immunology ; 122(2): 279-90, 2007 Oct.
Article in English | MEDLINE | ID: mdl-17848165

ABSTRACT

Dendritic cells (DC) play a critical role in initiating and directing adaptive immune responses against pathogens and tumours. Immature DC are thought to act as sentinels in peripheral tissues where their main function is to capture antigen at sites of infection, whereas mature DC are highly efficient at priming T-cell-mediated immune responses against infectious pathogens. The DC maturation process is thought to be an important step in the efficient generation of cytotoxic T lymphocytes (CTL). It is well established that many aspects of immune function, including CTL-mediated antiviral immunity, are modulated by neuroendocrine-derived products. Corticosterone (CORT), an adrenal hormone produced at increased concentrations during a stress response, has been shown to play a role in impaired CTL responses in stressed animals, leading to high mortality in mice normally resistant to viral infection. While direct effects of neuroendocrine mediators on CTL have been studied, little is known about their effects on DC that are critical for CTL priming. Here, we found that physiologically relevant concentrations of CORT, acting via the glucocorticoid receptor, functionally compromise DC maturation. DC exposed to CORT remained phenotypically and functionally immature after stimulation with lipopolysaccharide and were impaired for the production of interleukin (IL)-6, IL-12, and tumour necrosis factor-alpha. These effects were biologically significant, as CORT treatment resulted in a marked reduction in the ability of DC to prime naive CD8(+) T cells in vivo. These findings offer a potential mechanism underlying stress-associated immunosuppression.


Subject(s)
Corticosterone/pharmacology , Dendritic Cells/drug effects , Animals , Antigens, Surface/metabolism , CD8-Positive T-Lymphocytes/immunology , Cell Differentiation/drug effects , Cells, Cultured , Cytokines/biosynthesis , Dendritic Cells/immunology , Dendritic Cells/physiology , Endocytosis/drug effects , Histocompatibility Antigens Class II/metabolism , Immune Tolerance/drug effects , Lipopolysaccharides/antagonists & inhibitors , Male , Mice , Mice, Inbred C57BL , Receptors, Glucocorticoid/physiology , Up-Regulation/drug effects
5.
J Gen Virol ; 88(Pt 9): 2378-2386, 2007 Sep.
Article in English | MEDLINE | ID: mdl-17698646

ABSTRACT

Viral vectors have been shown to induce protective CD8(+) T-cell populations in animal models, but significant obstacles remain to their widespread use for human vaccination. One such obstacle is immunodominance, where the CD8(+) T-cell response to a vector can suppress the desired CD8(+) T-cell response to a recombinantly encoded antigen. To overcome this hurdle, we broadly reduced vector-specific gene expression. We treated a recombinant vaccinia virus, encoding antigen as a minimal peptide determinant (8-10 aa), with psoralen and short-wave UV light. The resulting virus induced 66 % fewer vector-specific immunodominant CD8(+) T cells, allowing the in vivo induction of an increased number of CD8(+) T cells specific for the recombinant antigen.


Subject(s)
CD8 Antigens/genetics , Gene Expression Regulation, Viral , Genetic Vectors , T-Lymphocytes/immunology , Animals , CD8-Positive T-Lymphocytes/immunology , Cytokines/analysis , Inflammation/etiology , Inflammation/prevention & control , Mice , Mice, Inbred C57BL , Transcription, Genetic , Ultraviolet Rays
6.
Cell Tissue Res ; 324(1): 1-8, 2006 Apr.
Article in English | MEDLINE | ID: mdl-16408195

ABSTRACT

We have previously described an immortal rat central-nervous-system progenitor cell line, AF5, which is able to exit the cell cycle and assume a differentiated state with neuronal properties. The phenotypic specification of differentiated AF5 cells, however, is not known. In the present study, when induced to differentiate by serum starvation in Neurobasal medium, AF5 cells down-regulate glial fibrillary acidic protein and up-regulate expression of beta-III-tubulin, medium-molecular-weight neurofilament protein, and neuronal growth-associated protein 43. Expression of the gamma-aminobutyric acid (GABA) lineage marker, glutamic acid decarboxylase 67 (GAD67), increases during differentiation, suggesting that AF5 cells adopt a GABAergic lineage. Time-course analysis of the GABAergic neuron specification transcription factor, Pitx2, by reverse transcription/polymerase chain reaction, has shown an increase in the Pitx2 transcript 48 h after initiation of differentiation. In differentiated AF5 cells, expression of the Pitx2 target gene products GAD65 and GABA transporter-1 increases. Cellular GABA levels in differentiated AF5 cells increase by about 26-fold, and GABA release into the medium is 150-fold higher compared with that of undifferentiated cells. Therefore, AF5 cells can be induced to differentiate to a neuronal phenotype with a GABAergic lineage.


Subject(s)
Cell Differentiation , Cell Lineage , Neurons/cytology , Stem Cells/cytology , gamma-Aminobutyric Acid/metabolism , Animals , Cells, Cultured , Culture Media/pharmacology , Gene Expression Regulation , Glutamate Decarboxylase/metabolism , Homeodomain Proteins/metabolism , Isoenzymes/metabolism , Rats , Transcription Factors/metabolism , gamma-Aminobutyric Acid/biosynthesis , Homeobox Protein PITX2
7.
J Neuroimmunol ; 160(1-2): 48-60, 2005 Mar.
Article in English | MEDLINE | ID: mdl-15710457

ABSTRACT

The presentation of viral peptide-MHC class I complexes by antigen presenting cells, such as dendritic cells (DCs), is obligatory for the generation of antiviral effector and memory CD8(+) cytotoxic T lymphocyte (CTL) responses. Prolonged psychological stress is immunosuppressive and undermines primary and memory CTL-mediated antiviral immunity; however, the mechanisms involved are unknown. Using a panel of novel reagents and techniques, we quantitatively measured the effect of the stress-induced hormone corticosterone (CORT) on the efficiency of DCs to process and present virally expressed antigen, characterized the conditions for this CORT-mediated effect, and delineated the components of the MHC class I pathway that were affected. We found that physiologically relevant levels of CORT, prior to infection and acting via the glucocorticoid receptor, suppressed the formation of peptide-MHC class I complexes on the surface of infected DCs. We further showed that this suppression of peptide-MHC class I complexes is via the action of CORT on elements of the class I pathway upstream from TAP that are involved in the generation of antigenic peptides. This CORT-mediated suppression of peptide-class I complexes on DCs also resulted in a marked reduction of their ability to activate a specific T cell hybridoma. These findings offer a mechanism contributing to the stress-induced suppression of host defenses against viral diseases and have implications for the efficacy of antiviral vaccines. At the most fundamental cellular level, this impairment of antigen processing has implications for the regulation of protein degradation in all cells, which is critical to many aspects of immune function.


Subject(s)
Antigen Presentation/drug effects , Corticosterone/pharmacology , Dendritic Cells/drug effects , Dendritic Cells/immunology , H-2 Antigens/immunology , Immunosuppressive Agents/pharmacology , Peptide Biosynthesis/drug effects , ATP Binding Cassette Transporter, Subfamily B, Member 2 , ATP Binding Cassette Transporter, Subfamily B, Member 3 , ATP-Binding Cassette Transporters/physiology , Animals , Antigen Presentation/immunology , Cell Line , Cell Membrane/drug effects , Cell Membrane/immunology , Cell Membrane/metabolism , Corticosterone/physiology , Dendritic Cells/metabolism , Dendritic Cells/virology , Dose-Response Relationship, Immunologic , Egg Proteins/antagonists & inhibitors , Egg Proteins/metabolism , Endoplasmic Reticulum/immunology , Endoplasmic Reticulum/metabolism , Epitopes/immunology , Epitopes/metabolism , H-2 Antigens/biosynthesis , H-2 Antigens/physiology , Hybridomas , Lymphocyte Activation/drug effects , Mice , Ovalbumin/antagonists & inhibitors , Ovalbumin/immunology , Ovalbumin/metabolism , Peptide Biosynthesis/immunology , Peptide Fragments/immunology , Peptide Fragments/metabolism , Protein Processing, Post-Translational/immunology , Receptors, Glucocorticoid/physiology , Signal Transduction/immunology , T-Lymphocytes/immunology , Vaccinia virus/immunology
8.
Expert Opin Biol Ther ; 4(6): 861-8, 2004 Jun.
Article in English | MEDLINE | ID: mdl-15174968

ABSTRACT

CD8(+) T cells (T(CD8+)) can mediate protective immunity to intracellular pathogens and tumours. Viruses generate strong T(CD8+) responses and, therefore, represent attractive vectors for generating vaccines aimed at producing T(CD8+)-mediated protective immunity. This review will examine the immunological properties of viruses that make them good candidates as vaccine vectors, as well as the manipulations of both vector and antigen that may be required to produce an effective vaccine. The areas addressed include virus infection of dendritic cells in vivo, stimulation of the innate immune response via intracellular and extracellular pattern recognition receptors, the effect of antigenic form on the pathways of antigen presentation and the requirement for elimination of viral genes that target various aspects of the innate and adaptive immune response.


Subject(s)
CD8 Antigens/immunology , CD8-Positive T-Lymphocytes/virology , Genetic Therapy/methods , Immune System/physiology , Immunotherapy/methods , Vaccines , Animals , Antigen Presentation , Antigen-Presenting Cells/metabolism , Antigens/chemistry , Dendritic Cells/metabolism , Genetic Vectors , Humans , Major Histocompatibility Complex , T-Lymphocytes/immunology
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