RESUMO
The heterodimeric cytokine interleukin 27 (IL-27) signals through the IL-27Rα subunit of its receptor, combined with gp130, a common receptor chain used by several cytokines, including IL-6. Notably, the IL-27 subunits p28 (IL-27p28) and EBI3 are not always expressed together, which suggests that they may have unique functions. Here we show that IL-27p28, independently of EBI3, antagonized cytokine signaling through gp130 and IL-6-mediated production of IL-17 and IL-10. Similarly, the ability to generate antibody responses was dependent on the activity of gp130-signaling cytokines. Mice transgenic for expression of IL-27p28 showed a substantial defect in the formation of germinal centers and antibody production. Thus, IL-27p28, as a natural antagonist of gp130-mediated signaling, may be useful as a therapeutic for managing inflammation mediated by cytokines that signal through gp130.
Assuntos
Receptor gp130 de Citocina/metabolismo , Interleucinas/metabolismo , Transdução de Sinais/imunologia , Animais , Formação de Anticorpos/imunologia , Linfócitos B/imunologia , Linfócitos B/metabolismo , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/metabolismo , Separação Celular , Receptor gp130 de Citocina/imunologia , Citocinas/imunologia , Citocinas/metabolismo , Ensaio de Imunoadsorção Enzimática , Feminino , Citometria de Fluxo , Imuno-Histoquímica , Interleucinas/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Antígenos de Histocompatibilidade Menor , Receptores de Citocinas/imunologia , Receptores de Citocinas/metabolismoRESUMO
To understand lymphocyte behavior in the brain, we used two-photon microscopy to visualize effector CD8(+) T cells during toxoplasmic encephalitis. These cells displayed multiple behaviors with two distinct populations of cells apparent: one with a constrained pattern of migration and one with a highly migratory subset. The proportion of these populations varied over time associated with changes in antigen availability as well as T cell expression of the inhibitory receptor PD1. Unexpectedly, the movement of infiltrating cells was closely associated with an infection-induced reticular system of fibers. This observation suggests that, whereas in other tissues pre-existing scaffolds exist that guide lymphocyte migration, in the brain specialized structures are induced by inflammation that guide migration of T cells in this immune-privileged environment.
Assuntos
Linfócitos T CD8-Positivos/citologia , Linfócitos T CD8-Positivos/imunologia , Movimento Celular/imunologia , Toxoplasma/imunologia , Toxoplasmose Cerebral/imunologia , Toxoplasmose Cerebral/parasitologia , Animais , Sistema Nervoso Central/imunologia , Camundongos , Ratos , Toxoplasmose Cerebral/patologiaRESUMO
The Toxoplasma gondii population consists of multiple strains, defined by genotype and virulence. Previous studies have established that protective immunity to this organism is mediated by IL-12, which drives T cells to produce IFN-gamma. Paradoxically, although type I and type II strains of T. gondii both induce IL-12 and IFN-gamma in the mouse, type I parasites are lethal, whereas type II strains establish chronic infection. The cellular basis for these strain-dependent differences remains unclear. To better understand these events, the CD8(+) T cell and dendritic cell (DC) responses to transgenic, OVA-expressing type I RH (RH OVA) and type II Prugniuad (Pru OVA) parasites were examined. Pru OVA-infected mice developed a robust DC response at the site of infection and the draining lymph node and generated a population of endogenous OVA-specific CD8(+) T cells. In contrast, RH OVA-infected mice had fewer DCs and OVA-specific CD8(+) T cells. RH OVA-infected mice given preactivated OVA-specific CD8(+) T cells were protected, suggesting that reduced DC-derived signals contributed to the low OVA-specific CD8(+) T cell numbers observed during type I infection. Indeed, DC depletion prior to Pru OVA infection resulted in a failure to generate activated OVA-specific CD8(+) T cells, and IL-12p70 treatment during RH OVA infection modestly increased the number of Ag-specific cells. Together, these data are consistent with a model of immunity to T. gondii in which strain-dependent DC responses shape the generation of Ag-specific CD8(+) T cells and determine the outcome of infection.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Células Dendríticas/imunologia , Células Dendríticas/parasitologia , Ativação Linfocitária/imunologia , Toxoplasma/imunologia , Toxoplasma/patogenicidade , Toxoplasmose Animal/imunologia , Animais , Linfócitos T CD8-Positivos/parasitologia , Linfócitos T CD8-Positivos/patologia , Proliferação de Células , Células Cultivadas , Células Dendríticas/patologia , Epitopos de Linfócito T/administração & dosagem , Epitopos de Linfócito T/genética , Epitopos de Linfócito T/imunologia , Feminino , Ativação Linfocitária/genética , Contagem de Linfócitos , Linfopenia/genética , Linfopenia/imunologia , Linfopenia/parasitologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Ovalbumina/administração & dosagem , Ovalbumina/biossíntese , Ovalbumina/genética , Toxoplasma/genética , Toxoplasmose Animal/genética , Toxoplasmose Animal/patologia , Virulência/imunologiaRESUMO
To better understand the initiation of CD8(+) T cell responses during infection, the primary response to the intracellular parasite Toxoplasma gondii was characterized using 2-photon microscopy combined with an experimental system that allowed visualization of dendritic cells (DCs) and parasite specific CD8(+) T cells. Infection with T. gondii induced localization of both these populations to the sub-capsular/interfollicular region of the draining lymph node and DCs were required for the expansion of the T cells. Consistent with current models, in the presence of cognate antigen, the average velocity of CD8(+) T cells decreased. Unexpectedly, infection also resulted in modulation of the behavior of non-parasite specific T cells. This TCR-independent process correlated with the re-modeling of the lymph node micro-architecture and changes in expression of CCL21 and CCL3. Infection also resulted in sustained interactions between the DCs and CD8(+) T cells that were visualized only in the presence of cognate antigen and were limited to an early phase in the response. Infected DCs were rare within the lymph node during this time frame; however, DCs presenting the cognate antigen were detected. Together, these data provide novel insights into the earliest interaction between DCs and CD8(+) T cells and suggest that cross presentation by bystander DCs rather than infected DCs is an important route of antigen presentation during toxoplasmosis.
Assuntos
Linfócitos T CD8-Positivos/patologia , Células Dendríticas/patologia , Microscopia de Fluorescência por Excitação Multifotônica/métodos , Toxoplasma/fisiologia , Toxoplasmose/patologia , Análise de Variância , Animais , Linfócitos T CD8-Positivos/metabolismo , Linfócitos T CD8-Positivos/parasitologia , Movimento Celular , Células Dendríticas/metabolismo , Células Dendríticas/parasitologia , Citometria de Fluxo , Cinética , Linfonodos/metabolismo , Camundongos , Camundongos Transgênicos , Toxoplasmose/metabolismoRESUMO
The nuclear factor κB transcription factor c-Rel is exclusively expressed in immune cells and plays a role in numerous cellular functions including proliferation, survival and production of chemokines and cytokines. c-Rel has also been implicated in the regulation of multiple genes involved in innate and adaptive immune responses to the intracellular protozoan parasite Toxoplasma gondii, in particular IL-12. To better understand how this transcription factor controls the CD8(+) T-cell response to this organism, wild-type (WT) and c-Rel(-/-) mice were challenged with a replication-deficient strain of T. gondii that expresses the model antigen ovalbumin (OVA). These studies revealed that c-Rel was required for optimal primary expansion of OVA-specific CD8(+) T cells and that immunized c-Rel-deficient mice were susceptible to challenge with a virulent strain of T. gondii. However, when c-Rel(-/-) cells specific for OVA were adoptively transferred into a WT recipient, or c-Rel(-/-) mice were treated with IL-12 at the time of immunization, there was no apparent proliferative defect. Surprisingly, upon secondary challenge, antigen-specific CD8(+) T cells in c-Rel(-/-) mice expanded to a much greater degree in terms of frequency as well as numbers when compared with WT mice. Despite this, the cytokine responses of c-Rel(-/-) mice remained defective, consistent with their susceptibility to secondary challenge. Together, these results indicate that in this infection model, the major influence of c-Rel in generation of CD8(+) T-cell responses is through its regulation of the inflammatory environment, rather than playing a substantial T-cell-intrinsic role.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/metabolismo , Proteínas Proto-Oncogênicas c-rel/metabolismo , Toxoplasma/imunologia , Animais , Camundongos , Camundongos Endogâmicos C57BL , Proteínas Proto-Oncogênicas c-rel/deficiênciaRESUMO
Multiple studies have established that the ability of CD8(+) T cells to act as cytolytic effectors and produce gamma interferon is important in mediating resistance to the intracellular parasite Toxoplasma gondii. To better understand the generation of the antigen-specific CD8(+) T-cell responses induced by T. gondii, mice were immunized with replication-deficient parasites that express the model antigen ovalbumin (OVA). Class I tetramers specific for SIINFEKL were used to track the OVA-specific endogenous CD8(+) T cells. The peak CD8(+) T-cell response was found at day 10 postimmunization, after which the frequency and numbers of antigen-specific cells declined. Unexpectedly, replication-deficient parasites were found to induce antigen-specific cells with faster kinetics than replicating parasites. The generation of optimal numbers of antigen-specific CD8(+) effector T cells was found to require CD4(+) T-cell help. At 7 days following immunization, antigen-specific cells were found to be CD62L(low), KLRG1(+), and CD127(low), and they maintained this phenotype for more than 70 days. Antigen-specific CD8(+) effector T cells in immunized mice exhibited potent perforin-dependent OVA-specific cytolytic activity in vivo. Perforin-dependent cytolysis appeared to be the major cytolytic mechanism; however, a perforin-independent pathway that was not mediated via Fas-FasL was also detected. This study provides further insight into vaccine-induced cytotoxic T-lymphocyte responses that correlate with protective immunity to T. gondii and identifies a critical role for CD4(+) T cells in the generation of protective CD8(+) T-cell responses.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Vacinas Protozoárias/imunologia , Toxoplasma/imunologia , Animais , Linfócitos T CD4-Positivos/fisiologia , Citotoxicidade Imunológica , Imunização , Imunofenotipagem , Cinética , Camundongos , Camundongos Endogâmicos C57BL , Ovalbumina/imunologiaRESUMO
Toxoplasma gondii is an important cause of clinical disease in fetuses, infants and immunocompromised patients. Since the discovery of T. gondii 100 years ago, this pathogen and the host's immune response to toxoplasmosis have been studied intensely. This has led to the development of a working model of immunity to T. gondii, and has also resulted in fundamental new insights into the role of various cytokines in resistance to infection. By examining this organism, researchers have identified many of the requirements for resistance to intracellular pathogens and characterized numerous regulatory factors, including interleukin-10 (IL-10) and IL-27, which control inflammatory processes. In the next 100 years of T. gondii immunobiology, researchers will have the opportunity to answer some of the long-standing questions in the field using new techniques and reagents. These future studies will be vital in building a more comprehensive model of immunity to this pathogen and in advancing our understanding of immunoregulation, particularly in humans. Ultimately, the challenge will be to use this information to develop new vaccines and therapies to manage disease in affected patients.
Assuntos
Citocinas/imunologia , Toxoplasma/imunologia , Toxoplasmose/imunologia , Animais , Humanos , Hospedeiro Imunocomprometido , Imunomodulação , Vacinas ProtozoáriasRESUMO
In this study, the role of NF-kappaB1 was examined during toxoplasmosis. While wildtype BALB/c mice generated protective responses, NF-kappaB1(-/-) mice developed Toxoplasmic encephalitis, characterized by increased parasite burden and necrosis in the brain. Susceptibility was primarily associated with a local decrease in the number of CD8(+) T cells and IFN-gamma production, while accessory cell function appeared intact in NF-kappaB1(-/-) mice. Consistent with these findings, T cell transfer studies revealed that NF-kappaB1(-/-) T cells provided SCID mice less protection than wildtype T cells. These results demonstrate an intrinsic role for NF-kappaB1 in T cell-mediated immunity to Toxoplasmagondii.
Assuntos
Imunidade Inata/imunologia , Subunidade p50 de NF-kappa B/metabolismo , Linfócitos T/imunologia , Toxoplasma/imunologia , Toxoplasmose Cerebral/imunologia , Animais , Encéfalo/imunologia , Encéfalo/parasitologia , Encéfalo/patologia , Linfócitos T CD8-Positivos/imunologia , Células Cultivadas , Imunidade Inata/genética , Interferon gama/metabolismo , Contagem de Linfócitos , Camundongos , Camundongos Knockout , Camundongos SCID , Subunidade p50 de NF-kappa B/genética , RNA Mensageiro/metabolismo , Toxoplasmose Cerebral/genética , Toxoplasmose Cerebral/fisiopatologiaRESUMO
Toxoplasma gondii is an important cause of clinical disease in fetuses, infants and immunocompromised patients. Since the discovery of T. gondii 100 years ago, this pathogen and the host's immune response to toxoplasmosis have been studied intensely. This has led to the development of a working model of immunity to T. gondii, and has also resulted in fundamental new insights into the role of various cytokines in resistance to infection. By examining this organism, researchers have identified many of the requirements for resistance to intracellular pathogens and characterized numerous regulatory factors, including interleukin-10 (IL-10) and IL-27, which control inflammatory processes. In the next 100 years of T. gondii immunobiology, researchers will have the opportunity to answer some of the long-standing questions in the field using new techniques and reagents. These future studies will be vital in building a more comprehensive model of immunity to this pathogen and in advancing our understanding of immunoregulation, particularly in humans. Ultimately, the challenge will be to use this information to develop new vaccines and therapies to manage disease in affected patients.