RESUMO
Inhibitors of apoptosis proteins (IAPs) were originally described as regulating apoptosis by direct binding to caspases. More recently, IAPs have been identified as important modulators of canonical and noncanonical nuclear factor κB signaling via their ubiquitin-E3 ligase activity. IAPs are therefore, not only gatekeepers of cell death, but are probably also involved in the regulation of inflammation, as well as innate and adaptive immunity. In this study, we analyzed the role of IAPs in T-cell immunity during lymphocytic choriomeningitis virus (LCMV) infection by pharmacological targeting with an IAP antagonist/second mitochondria-derived activator of caspase-mimetic. Expansion of virus-specific CD8 T cells was drastically reduced in LCMV-infected mice exposed to IAP antagonists. Accordingly, virus control was substantially impaired, indicated by high virus titres in the spleen and the spread of LCMV to peripheral organs. The profound negative effect of IAP antagonists on T-cell immunity was partially linked to tumor necrosis factor-mediated cell death of activated T cells and required inhibition of X-linked inhibitor of apoptosis, as well as cellular IAP-1. Thus, IAPs play an important role in T-cell expansion and survival in the context of a highly inflammatory environment such as a virus infection, indicating that IAP antagonists may interfere with immune responses.
Assuntos
Proteínas Inibidoras de Apoptose/imunologia , Coriomeningite Linfocítica/imunologia , Vírus da Coriomeningite Linfocítica/imunologia , Linfócitos T/imunologia , Linfócitos T/virologia , Animais , Linfócitos T CD4-Positivos/citologia , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/citologia , Linfócitos T CD8-Positivos/imunologia , Sobrevivência Celular , Imunidade , Proteínas Inibidoras de Apoptose/antagonistas & inibidores , Ativação Linfocitária , Camundongos , Camundongos Endogâmicos C57BL , NF-kappa B/imunologia , Linfócitos T/citologia , Fatores de Necrose Tumoral/imunologia , Proteínas Inibidoras de Apoptose Ligadas ao Cromossomo X/antagonistas & inibidores , Proteínas Inibidoras de Apoptose Ligadas ao Cromossomo X/imunologiaRESUMO
Trogocytosis describes the transfer of surface determinants between immune cells and has been implicated in immune regulation. Most findings are based on in vitro studies since in vivo trogocytosis of immune cells is difficult to detect under physiological conditions. We used low frequencies of memory P14 T cells to demonstrate that T cells perform trogocytosis in vivo if in contact with APC pulsed with GP33-peptide or expressing the antigen endogenously. Furthermore, in vivo trogocytosis of T cells is demonstrated during infections with lymphocytic choriomeningitis virus and vaccinia virus. Trogocytosis-positive T cells revealed higher expression of activation marker and cytokines, showing a more activated phenotype compared to trogocytosis-negative T cells.
Assuntos
Células Apresentadoras de Antígenos/metabolismo , Infecções por Arenaviridae/imunologia , Vírus da Coriomeningite Linfocítica/imunologia , Fusão de Membrana , Linfócitos T/metabolismo , Transferência Adotiva , Animais , Células Apresentadoras de Antígenos/imunologia , Células Apresentadoras de Antígenos/patologia , Antígenos de Superfície/metabolismo , Antígenos Virais/imunologia , Células Cultivadas , Glicoproteínas/imunologia , Ativação Linfocitária , Vírus da Coriomeningite Linfocítica/patogenicidade , Fusão de Membrana/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Fragmentos de Peptídeos/imunologia , Receptores de Antígenos de Linfócitos T/genética , Linfócitos T/imunologia , Linfócitos T/patologia , Linfócitos T/virologia , Proteínas Virais/imunologiaRESUMO
To study the role of IL-12 as a third signal for T-cell activation and differentiation in vivo, direct IL-12 signaling to CD8(+) T cells was analyzed in bacterial and viral infections using the P14 T-cell adoptive transfer model with CD8(+) T cells that lack the IL-12 receptor. Results indicate that CD8(+) T cells deficient in IL-12 signaling were impaired in clonal expansion after Listeria monocytogenes infection but not after infection with lymphocytic choriomeningitis virus, vaccinia virus or vesicular stomatitis virus. Although limited in clonal expansion after Listeria infection, CD8(+) T cells deficient in IL-12 signaling exhibited normal degranulation activity, cytolytic functions, and secretion of IFN-gamma and TNF-alpha. However, CD8(+) T cells lacking IL-12 signaling failed to up-regulate KLRG1 and to down-regulate CD127 in the context of Listeria but not viral infections. Thus, direct IL-12 signaling to CD8(+) T cells determines the cell fate decision between short-lived effector cells and memory precursor effector cells, which is dependent on pathogen-induced local cytokine milieu.
Assuntos
Diferenciação Celular/imunologia , Listeriose/imunologia , Coriomeningite Linfocítica/imunologia , Linfócitos T/citologia , Transferência Adotiva/métodos , Animais , Linfócitos T CD8-Positivos/citologia , Linfócitos T CD8-Positivos/microbiologia , Linfócitos T CD8-Positivos/virologia , Linhagem Celular , Linhagem Celular Tumoral , Proliferação de Células , Citometria de Fluxo , Memória Imunológica/imunologia , Interferon gama/imunologia , Subunidade alfa de Receptor de Interleucina-7/imunologia , Lectinas Tipo C , Listeriose/microbiologia , Listeriose/terapia , Coriomeningite Linfocítica/virologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Receptores Imunológicos/imunologia , Receptores de Interleucina-12/deficiência , Receptores de Interleucina-12/genética , Receptores de Interleucina-12/imunologia , Transdução de Sinais/imunologia , Linfócitos T/microbiologia , Linfócitos T/virologia , Fatores de Tempo , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Signal 3 cytokines, such as IL-12 or type I IFN, support expansion and differentiation of CD8 T cells in vivo. If and how these two signal 3 cytokines compensate each other in T cell activation during different infections is so far unknown. Using CD8 T cells lacking receptors for IL-12, type I IFN or both, we show that the expansion of CD8 T cells depends on type I IFN (LCMV infection), type I IFN and IL-12 (Listeria and vesicular stomatitis virus infection) or is largely independent of the two cytokines (vaccinia virus infection). Furthermore, we show that CD8 T cells lacking IL-12 and type I IFN signals are impaired in cytokine production and cytolytic activity in the context of VSV and Listeria infection. These effector CD8 T cells fail to express KLRG1, thereby exhibiting a memory-like phenotype which correlated with lower expression of the transcription factor T-bet and higher expression of Eomes. This indicates that the variable interplay of both signal 3 cytokines is mandatory for cell fate decision of CD8 T cells in the context of different infections. Furthermore our results demonstrate that the pathogen-induced overall inflammatory milieu and not the antigen load and/or the quality of antigen presentation critically determine the signal 3 dependence of CD8 T cells.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Imunidade/imunologia , Infecções/imunologia , Interferon Tipo I/imunologia , Interleucina-12/imunologia , Animais , Proliferação de Células , Infecções/microbiologia , Infecções/patologia , Infecções/virologia , Inflamação/imunologia , Inflamação/patologia , Listeriose/imunologia , Listeriose/patologia , Ativação Linfocitária/imunologia , Vírus da Coriomeningite Linfocítica/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Proteínas com Domínio T/metabolismo , Vesiculovirus/imunologiaRESUMO
The role of type I IFN signaling in CD8 T cells was analyzed in an adoptive transfer model using P14 TCR transgenic CD8 T cells specific for lymphocytic choriomeningitis virus (LCMV) but deficient in type I IFNR. In the present study, we demonstrate severe impairment in the capacity of P14 T cells lacking type I IFNR to expand in normal type I IFNR wild-type C57BL/6 hosts after LCMV infection. In contrast, following infection of recipient mice with recombinant vaccinia virus expressing LCMV glycoprotein, P14 T cell expansion was considerably less dependent on type I IFNR expression. Lack of type I IFNR expression by P14 T cells did not affect cell division after LCMV infection but interfered with clonal expansion. Thus, direct type I IFN signaling is essential for CD8 T cell survival in certain viral infections.