RESUMEN
An estimated one-third of the world's population is infected with Mycobacterium tuberculosis, although most affected individuals maintain a latent infection. This control is attributed to the formation of granulomas, cell masses largely comprising infected macrophages with T cells aggregated around them. Inflammatory DCs, characterized as CD11c+CD11b+Ly6C+, are also found in granulomas and are an essential component of the acute immune response to mycobacteria. However, their function during chronic infection is less well understood. Here, we report that CD11c+ cells dynamically traffic in and out of both acute and chronic granulomas induced by Mycobacterium bovis strain bacillus Calmette-Guérin (BCG) in mice. By transplanting Mycobacterium-induced granulomas containing fluorescently labeled CD11c+ cells and bacteria into unlabeled mice, we were able to follow CD11c+ cell trafficking and T cell activation. We found that half of the CD11c+ cells in chronic granulomas were exchanged within 1 week. Compared with tissue-resident DC populations, CD11c+ cells migrating out of granuloma-containing tissue had an unexpected systemic dissemination pattern. Despite low antigen availability, systemic CD4+ T cell priming still occurred during chronic infection. These data demonstrate that surveillance of granulomatous tissue by CD11c+ cells is continuous and that these cells are distinct from tissue-resident DC populations and support T cell priming during both stages of Mycobacterium infection. This intense DC surveillance may also be a feature of Mycobacterium tuberculosis infection and other granuloma-associated diseases.
Asunto(s)
Células Dendríticas/inmunología , Células Dendríticas/patología , Granuloma/inmunología , Granuloma/patología , Infecciones por Mycobacterium/inmunología , Infecciones por Mycobacterium/patología , Animales , Antígeno CD11c/metabolismo , Movimiento Celular/inmunología , Movimiento Celular/fisiología , Células Dendríticas/fisiología , Modelos Animales de Enfermedad , Granuloma/microbiología , Humanos , Activación de Linfocitos , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Infecciones por Mycobacterium/microbiología , Mycobacterium bovis/inmunología , Mycobacterium bovis/patogenicidad , Linfocitos T/inmunología , Linfocitos T/microbiología , Linfocitos T/patología , Tuberculosis/inmunología , Tuberculosis/microbiología , Tuberculosis/patologíaRESUMEN
The NLR gene family mediates host immunity to various acute pathogenic stimuli, but its role in chronic infection is not known. This paper addressed the role of NLRP3 (NALP3), its adaptor protein PYCARD (ASC), and caspase-1 during infection with Mycobacterium tuberculosis (Mtb). Mtb infection of macrophages in culture induced IL-1beta secretion, and this requires the inflammasome components PYCARD, caspase-1, and NLRP3. However, in vivo Mtb aerosol infection of Nlrp3(-/-), Casp-1(-/-), and WT mice showed no differences in pulmonary IL-1beta production, bacterial burden, or long-term survival. In contrast, a significant role was observed for Pycard in host protection during chronic Mtb infection, as shown by an abrupt decrease in survival of Pycard(-/-) mice. Decreased survival of Pycard(-/-) animals was associated with defective granuloma formation. These data demonstrate that PYCARD exerts a novel inflammasome-independent role during chronic Mtb infection by containing the bacteria in granulomas.
Asunto(s)
Proteínas Portadoras/metabolismo , Caspasa 1/metabolismo , Proteínas del Citoesqueleto/metabolismo , Granuloma/microbiología , Granuloma/patología , Mycobacterium tuberculosis/fisiología , Tuberculosis/inmunología , Animales , Proteínas Reguladoras de la Apoptosis , Proteínas Adaptadoras de Señalización CARD , Células Cultivadas , Enfermedad Crónica , Proteínas del Citoesqueleto/deficiencia , Femenino , Humanos , Interleucina-1beta/biosíntesis , Interleucina-1beta/metabolismo , Macrófagos/metabolismo , Macrófagos/microbiología , Ratones , Mycobacterium tuberculosis/patogenicidad , Proteína con Dominio Pirina 3 de la Familia NLR , Tuberculosis/metabolismoRESUMEN
BACKGROUND: Mycobacterium-induced granulomas are the interface between bacteria and host immune response. During acute infection dendritic cells (DCs) are critical for mycobacterial dissemination and activation of protective T cells. However, their role during chronic infection in the granuloma is poorly understood. METHODOLOGY/PRINCIPAL FINDINGS: We report that an inflammatory subset of murine DCs are present in granulomas induced by Mycobacteria bovis strain Bacillus Calmette-guerin (BCG), and both their location in granulomas and costimulatory molecule expression changes throughout infection. By flow cytometric analysis, we found that CD11c(+) cells in chronic granulomas had lower expression of MHCII and co-stimulatory molecules CD40, CD80 and CD86, and higher expression of inhibitory molecules PD-L1 and PD-L2 compared to CD11c(+) cells from acute granulomas. As a consequence of their phenotype, CD11c(+) cells from chronic lesions were unable to support the reactivation of newly-recruited, antigen 85B-specific CD4(+)IFNgamma(+) T cells or induce an IFNgamma response from naïve T cells in vivo and ex vivo. The mechanism of this inhibition involves the PD-1:PD-L signaling pathway, as ex vivo blockade of PD-L1 and PD-L2 restored the ability of isolated CD11c(+) cells from chronic lesions to stimulate a protective IFNgamma T cell response. CONCLUSIONS/SIGNIFICANCE: Our data suggest that DCs in chronic lesions may facilitate latent infection by down-regulating protective T cell responses, ultimately acting as a shield that promotes mycobacterium survival. This DC shield may explain why mycobacteria are adapted for long-term survival in granulomatous lesions.
Asunto(s)
Células Dendríticas/inmunología , Granuloma/inmunología , Linfocitos T/microbiología , Animales , Antígenos Bacterianos/inmunología , Antígenos CD/metabolismo , Antígeno B7-1/metabolismo , Antígeno B7-2/metabolismo , Antígeno B7-H1 , Antígeno CD11c/metabolismo , Linfocitos T CD4-Positivos/metabolismo , Antígenos CD40/metabolismo , Células Cultivadas , Células Dendríticas/metabolismo , Citometría de Flujo , Granuloma/metabolismo , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Interferón gamma/metabolismo , Ratones , Ratones Endogámicos C57BL , Microscopía Fluorescente , Infecciones por Mycobacterium/inmunología , Mycobacterium bovis/inmunología , Proteína 2 Ligando de Muerte Celular Programada 1 , Linfocitos T/metabolismoRESUMEN
Mycobacterium bovis BCG is still the most widely used vaccine against tuberculosis and CD8(+) T cells play important roles in fighting infection. We investigated how well antigen is processed and presented to CD8(+) T cells using the same well-characterized CD8(+) T cell epitope SIINFEKL expressed in either a cytoplasmic (GFP-OVA) or secreted (85B-OVA) context from BCG. We report that secreted SIINFEKL from 85B-OVA BCG is presented better than cytoplasmic SIINFEKL expressed by GFP-OVA BCG.