RESUMO
Infection of mice with the bacterium Ehrlichia muris elicits a protective T cell-independent (TI) IgM response mediated primarily by a population of CD11c-expressing plasmablasts in the spleen. Although splenic marginal zone (MZ) B cells are considered to be important for TI responses to blood-borne pathogens, MZ B cells were not responsible for generating plasmablasts in response to Ehrlichia muris. Moreover, antigen-specific serum IgM was decreased only modestly in splenectomized mice and in mice that lacked spleen, lymph nodes, and Peyer's patches (SLP mice). Both splenectomized and SLP mice were protected against lethal ehrlichial challenge infection. Moreover, we found a high frequency of Ehrlichia-specific plasmablasts in the omentum of both conventional and SLP mice. Omental plasmablasts elicited during Ehrlichia infection lacked expression of CD138 but expressed CD11c in a manner similar to that of their splenic counterparts. Selective ablation of CD11c-expressing B cells nearly eliminated the omental Ehrlichia-specific plasmablasts and reduced antigen-specific serum IgM, identifying the omental B cells as a source of IgM production in the SLP mice. Generation of the omental plasmablasts was route dependent, as they were detected following peritoneal infection but not following intravenous infection. Our data identify the omentum as an important auxiliary site of IgM production during intracellular bacterial infection.
Assuntos
Anticorpos Antibacterianos/metabolismo , Ehrlichiose/imunologia , Imunoglobulina M/metabolismo , Omento/imunologia , Plasmócitos/imunologia , Animais , Antígeno CD11c/análise , Ehrlichia/imunologia , Feminino , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Omento/patologia , Sindecana-1/análiseRESUMO
Fully functional T cells are necessary for the maintenance of protective immunity during chronic infections. However, activated T cells often undergo apoptosis or exhaustion upon chronic stimulation mediated by Ag or inflammation. T cell attrition can be compensated for by the production of thymus-derived T cells, although the new naive T cells must undergo T cell priming and differentiation under conditions different from those encountered during acute infection. We used a murine model of Mycobacterium tuberculosis infection to address how the activation and differentiation of new thymic emigrants is affected by chronic inflammation, as well as whether the newly developed effector T cells help to maintain peripheral T cell responses. Although new thymic emigrants contributed to the peripheral T cell response early during acute M. tuberculosis infection, the relative contribution of new effector T cells to the peripheral CD4 and CD8 T cell pools declined during chronic infection. The decline in new T cell recruitment was a consequence of quantitative and/or qualitative changes in Ag presentation, because during chronic infection both the priming and expansion of naive T cells were inefficient. Thus, although thymic tolerance is not a major factor that limits protective T cell responses, the chronic environment does not efficiently support naive T cell priming and accumulation during M. tuberculosis infection. These studies support our previous findings that long-term protective T cell responses can be maintained indefinitely in the periphery, but also suggest that the perturbation of homeostasis during chronic inflammatory responses may elicit immune pathology mediated by new T cells.
Assuntos
Mycobacterium tuberculosis/imunologia , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismo , Tuberculose Pulmonar/imunologia , Tuberculose Pulmonar/metabolismo , Animais , Modelos Animais de Doenças , Camundongos , Camundongos Congênicos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Subpopulações de Linfócitos T/microbiologia , Timo/citologia , Timo/imunologia , Tuberculose Pulmonar/patologiaRESUMO
IgM responses are well known to occur early postinfection and tend to be short-lived, which has suggested that this Ig does not significantly contribute to long-term immunity. In this study, we demonstrate that chronic infection with the intracellular bacterium Ehrlichia muris elicits a protective, long-term IgM response. Moreover, we identified a population of CD138(high)IgM(high) B cells responsible for Ag-specific IgM production in the bone marrow. The IgM-secreting cells, which exhibited characteristics of both plasmablasts and plasma cells, contributed to protection against fatal ehrlichial challenge. Mice deficient in activation-induced cytidine deaminase, which produce only IgM, were protected against fatal ehrlichial challenge infection. The IgM-secreting cells that we have identified were maintained in the bone marrow in the absence of chronic infection, as antibiotic-treated mice remained protected against challenge infection. Our studies identify a cell population that is responsible for the IgM production in the bone marrow, and they highlight a novel role for IgM in the maintenance of long-term immunity during intracellular bacterial infection.
Assuntos
Células da Medula Óssea/imunologia , Células da Medula Óssea/microbiologia , Ehrlichiose/imunologia , Ehrlichiose/prevenção & controle , Imunoglobulina M/biossíntese , Líquido Intracelular/imunologia , Plasmócitos/imunologia , Plasmócitos/microbiologia , Animais , Células da Medula Óssea/metabolismo , Doença Crônica , Ehrlichia/imunologia , Ehrlichiose/microbiologia , Imunoglobulina M/fisiologia , Líquido Intracelular/microbiologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Plasmócitos/metabolismo , Células Precursoras de Linfócitos B/imunologia , Células Precursoras de Linfócitos B/metabolismo , Células Precursoras de Linfócitos B/microbiologia , Baço/imunologia , Baço/metabolismo , Baço/microbiologia , Fatores de TempoRESUMO
The immune response elicited after Mycobacterium tuberculosis (Mtb) infection is critically dependent on CD4 T cells during both acute and chronic infection. How CD4 T-cell responses are maintained throughout infection is not well understood, and evidence from other infection models has suggested that, under conditions of chronic antigen stimulation, T cells can undergo replicative exhaustion. These findings led us to determine whether subpopulations of CD4 T cells existed that displayed markers of terminal differentiation or exhaustion during murine Mtb infection. Analysis of antigen-specific effector CD4 T cells revealed that programmed death-1 (PD-1) and the killer cell lectin-like receptor G1 (KLRG1) delineated subpopulations of T cells. PD-1-expressing CD4 T cells were highly proliferative, whereas KLRG1 cells exhibited a short lifespan and secreted the cytokines IFNγ and TNFα. Adoptive transfer studies demonstrated that proliferating PD-1-positive CD4 T cells differentiated into cytokine-secreting KLRG1-positive T cells, but not vice versa. Thus, proliferating PD-1-positive cells are not exhausted, but appear to be central to maintaining antigen-specific effector T cells during chronic Mtb infection. Our findings suggest that antigen-specific T-cell responses are maintained during chronic mycobacterial infection through the continual production of terminal effector cells from a proliferating precursor population.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Mycobacterium tuberculosis/imunologia , Especificidade do Receptor de Antígeno de Linfócitos T/imunologia , Tuberculose/imunologia , Animais , Antígenos de Bactérias/imunologia , Antígenos de Superfície , Proteínas Reguladoras de Apoptose , Linfócitos T CD4-Positivos/citologia , Diferenciação Celular/imunologia , Proliferação de Células , Senescência Celular/imunologia , Citocinas/metabolismo , Lectinas Tipo C , Camundongos , Receptor de Morte Celular Programada 1 , Receptores ImunológicosRESUMO
After respiratory virus infections, memory CD8+ T cells are maintained in the lung airways by a process of continual recruitment. Previous studies have suggested that this process is controlled, at least in the initial weeks after virus clearance, by residual antigen in the lung-draining mediastinal lymph nodes (MLNs). We used mouse models of influenza and parainfluenza virus infection to show that intranasally (i.n.) primed memory CD8+ T cells possess a unique ability to be reactivated by residual antigen in the MLN compared with intraperitoneally (i.p.) primed CD8+ T cells, resulting in the preferential recruitment of i.n.-primed memory CD8+ T cells to the lung airways. Furthermore, we demonstrate that the inability of i.p.-primed memory CD8+ T cells to access residual antigen can be corrected by a subsequent i.n. virus infection. Thus, two independent factors, initial CD8+ T cell priming in the MLN and prolonged presentation of residual antigen in the MLN, are required to maintain large numbers of antigen-specific memory CD8+ T cells in the lung airways.
Assuntos
Antígenos Virais/imunologia , Linfócitos T CD8-Positivos/virologia , Apresentação Cruzada/imunologia , Memória Imunológica/imunologia , Pulmão/imunologia , Orthomyxoviridae/imunologia , Vírus Sendai/imunologia , Administração Intranasal , Animais , Linfócitos T CD8-Positivos/citologia , Linfócitos T CD8-Positivos/imunologia , Movimento Celular , Epitopos , Injeções Intraperitoneais , Pulmão/virologia , Linfonodos/imunologia , Linfonodos/virologia , Mediastino/virologia , Camundongos , Especificidade da EspécieRESUMO
CD4(+) T cell responses to aerosol Mycobacterium tuberculosis (Mtb) infection are characterized by the relatively delayed appearance of effector T cells in the lungs. This delay in the adaptive response is likely critical in allowing the bacteria to establish persistent infection. Because of limitations associated with the detection of low frequencies of naïve T cells, it had not been possible to precisely determine when and where naïve antigen-specific T cells are first activated. We have addressed this problem by using early secreted antigenic target 6 (ESAT-6)-specific transgenic CD4 T cells to monitor early T cell activation in vivo. By using an adoptive transfer approach, we directly show that T cell priming to ESAT-6 occurs only after 10 days of infection, is initially restricted to the mediastinal lymph nodes, and does not involve other lymph nodes or the lungs. Primed CD4 T cells rapidly differentiated into proliferating effector cells and ultimately acquired the ability to produce IFN-gamma and TNF-alpha ex vivo. Initiation of T cell priming was enhanced by two full days depending on the magnitude of the challenge inoculum, which suggests that antigen availability is a factor limiting the early CD4 T cell response. These data define a key period in the adaptive immune response to Mtb infection.
Assuntos
Antígenos de Bactérias/imunologia , Proteínas de Bactérias/imunologia , Linfócitos T CD4-Positivos/imunologia , Linfonodos/imunologia , Mediastino , Mycobacterium tuberculosis/imunologia , Tuberculose/imunologia , Transferência Adotiva , Animais , Antígenos de Bactérias/genética , Proteínas de Bactérias/genética , Citometria de Fluxo , Cinética , Ativação Linfocitária/imunologia , Camundongos , Camundongos TransgênicosRESUMO
CD8+ T cells are a major source of IFN-gamma, a key effector cytokine in immune responses against many viruses and protozoa. Although the transcription factor T-bet is required for IFN-gamma expression in CD4+ T cells, it is reportedly dispensable in CD8+ T cells, where the transcription factor Eomesodermin is thought to be sufficient. The diverse functions of IFN-gamma are mediated through the IFN-gammaR and STAT1. In CD4+ T cells, STAT1 appears to be critical for the activation of T-bet and IFN-gamma, suggesting an IFN-gamma-dependent positive feedback loop. However, STAT1 can also be activated by other cytokines, including IL-27. In the present study we show that, in contrast to in vitro conditions and the prevailing paradigm, T-bet is critical for the in vivo IFN-gamma production by CD8+ T cells upon infection of mice with diverse pathogens. Whereas IFN-gammaR signals are dispensable for the T-bet-dependent IFN-gamma production, direct IL-27Ralpha signals are critical.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Infecções/imunologia , Interferon gama/metabolismo , Receptores de Citocinas/fisiologia , Proteínas com Domínio T/fisiologia , Animais , Humanos , Infecções/microbiologia , Infecções/parasitologia , Influenza Humana/imunologia , Interferon gama/genética , Camundongos , Camundongos Mutantes , Receptores de Citocinas/genética , Receptores de Interleucina , Fator de Transcrição STAT1/metabolismo , Proteínas com Domínio T/genética , Toxoplasmose/imunologiaRESUMO
Increasing age is associated with the development of CD8+ T cell clonal expansions (TCE) that can dominate the peripheral T cell repertoire and interfere with immune responses to infection and vaccination. Some TCE are driven by chronic infections, consistent with dysregulated outgrowth of T cell clones in response to persistent antigenic stimulation. However, a second class of TCE develops with age in the absence of chronic infections and is poorly understood in terms of origin or Ag dependence. In this study, we present evidence that Ag-specific TCE develop at high frequencies from conventional memory CD8+ T cell pools elicited by nonpersistent influenza and parainfluenza virus infections. Putative TCE occurred in both the central- and effector-memory CD8+ T cell populations and did not require Ag for their maintenance. In addition, they were similar to normal memory T cells in terms of phenotype and function, suggesting that they develop stochastically from the memory T cell pool. These data suggest that memory T cell pools become progressively dysregulated over time and this may have a significant impact on immune responsiveness in the aged.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Epitopos de Linfócito T/imunologia , Vírus da Influenza A Subtipo H3N2/imunologia , Ativação Linfocitária/imunologia , Infecções Respiratórias/imunologia , Vírus Sendai/imunologia , Subpopulações de Linfócitos T/imunologia , Doença Aguda , Envelhecimento/imunologia , Animais , Linfócitos T CD8-Positivos/patologia , Linfócitos T CD8-Positivos/virologia , Proliferação de Células , Células Clonais , Epitopos de Linfócito T/biossíntese , Feminino , Contagem de Linfócitos , Camundongos , Camundongos Endogâmicos C57BL , Infecções por Orthomyxoviridae/imunologia , Infecções por Orthomyxoviridae/patologia , Receptores de Antígenos de Linfócitos T alfa-beta/biossíntese , Infecções Respiratórias/patologia , Infecções por Respirovirus/imunologia , Infecções por Respirovirus/patologia , Subpopulações de Linfócitos T/patologia , Subpopulações de Linfócitos T/virologiaRESUMO
The contributions of different subsets of memory CD8+ T cells to recall responses at mucosal sites of infection are poorly understood. Here, we analyzed the CD8+ T cell recall responses to respiratory virus infection in mice and demonstrate that activation markers, such as CD27 and CD43, define three distinct subpopulations of memory CD8+ T cells that differ in their capacities to mount recall responses. These subpopulations are distinct from effector- and central-memory subsets, coordinately express other markers associated with activation status, including CXCR3, CD127, and killer cell lectin-like receptor G1, and are superior to CD62L in predicting the capacity of memory T cells to mediate recall responses. Furthermore, the capacity of vaccines to elicit these memory T cell subpopulations predicted the efficacy of the recall response. These findings extend our understanding of how recall responses are generated and suggest that activation and migration markers define distinct, and unrelated, characteristics of memory T cells.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Memória Imunológica , Ativação Linfocitária , Infecções por Orthomyxoviridae/imunologia , Infecções Respiratórias/imunologia , Linfócitos T/imunologia , Animais , Células Matadoras Naturais/imunologia , Leucossialina/imunologia , Camundongos , Fenótipo , Infecções Respiratórias/virologia , Subpopulações de Linfócitos T/imunologiaRESUMO
Infection with Mycobacterium bovis Bacille Calmette Guérin (BCG) induces high levels of apoptosis among activated CD4+ T cells. We have investigated the specificity of this pro-apoptotic response and its influence on CD4+ T cell mediated autoimmunity. Apoptosis induced by BCG-infection is unrelated to antigenic specificity, as demonstrated by the increased apoptosis of activated TCR transgenic CD4+ T cells of unrelated specificity. Moreover, infection-induced apoptosis promoted the deletion of CD4+ T cells activated either by peptide or anti-CD3/anti-CD28 stimulation. Infection-induced apoptosis required IFN-gamma production by the infected host, and expression of the IFN-gamma receptor on donor CD4+ T cells. We used an adoptive transfer model of experimental autoimmune encephalomyelitis (EAE) to assess the influence of infection-induced apoptosis on a CD4+ T cell-mediated response. A significantly higher level of apoptosis was seen among sorted encephalitogenic CD4+ T cells transferred to BCG-infected versus uninfected hosts. BCG-infected mice displayed a milder course of clinical disease than their uninfected counterparts and a decreased recovery of donor cells from the CNS. The data suggest that mycobacterial infection attenuates the severity of EAE, at least in part, by promoting the apoptotic elimination of autoreactive CD4+ T cells.
Assuntos
Apoptose , Autoimunidade/imunologia , Efeito Espectador/imunologia , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/microbiologia , Tuberculose/imunologia , Tuberculose/microbiologia , Transferência Adotiva , Animais , Antígenos/imunologia , Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD4-Positivos/patologia , Encefalomielite Autoimune Experimental/imunologia , Encefalomielite Autoimune Experimental/microbiologia , Interferon gama/genética , Interferon gama/imunologia , Interferon gama/metabolismo , Camundongos , Camundongos Knockout , Mycobacterium bovis/imunologia , Mycobacterium bovis/fisiologia , Receptores de Antígenos de Linfócitos T/genética , Receptores de Antígenos de Linfócitos T/imunologia , Receptores de Antígenos de Linfócitos T/metabolismo , Tuberculose/metabolismo , Tuberculose/patologiaRESUMO
Experimental autoimmune encephalomyelitis (EAE) disease was accelerated iNOS-deficient (KO) mice: coinciding with greatly increased numbers of Ag-specific Th1 cells in the periphery that appeared to rapidly shift from the spleen to the CNS during onset of disease symptoms. iNOS KO mice had significantly increased Th1 cells in the CNS versus wild-type mice. Apoptosis of CNS-infiltrating CD4(+) T cells was impaired in iNOS KO mice at peak of disease; consequently, these mice had more CNS-infiltrating CD4(+) T cells. Subsequently, iNOS KO mice up-regulated apoptosis of CNS-CD4(+) T cells. During chronic EAE, CNS macrophages were greatly decreased, suggesting elimination of CNS-infiltrating CD4(+) T cells and activated macrophages by iNOS-independent mechanisms. INOS is not only required for apoptosis of CNS-CD4(+) T cells but also prevents overexpansion of autoreactive Th1 cells in the periphery and the CNS.