RESUMEN
Alternatively activated M2 macrophages play an important role in maintenance of tissue homeostasis by scavenging dead cells, cell debris and lipoprotein aggregates via phagocytosis. Using proteomics, we investigated how alternative activation, driven by IL-4, modulated the phagosomal proteome to control macrophage function. Our data indicate that alternative activation enhances homeostatic functions such as proteolysis, lipolysis and nutrient transport. Intriguingly, we identified the enhanced recruitment of the TAK1/MKK7/JNK signalling complex to phagosomes of IL-4-activated macrophages. The recruitment of this signalling complex was mediated through K63 polyubiquitylation of the macrophage scavenger receptor 1 (MSR1). Triggering of MSR1 in IL-4-activated macrophages leads to enhanced JNK activation, thereby promoting a phenotypic switch from an anti-inflammatory to a pro-inflammatory state, which was abolished upon MSR1 deletion or JNK inhibition. Moreover, MSR1 K63 polyubiquitylation correlated with the activation of JNK signalling in ovarian cancer tissue from human patients, suggesting that it may be relevant for macrophage phenotypic shift in vivo Altogether, we identified that MSR1 signals through JNK via K63 polyubiquitylation and provides evidence for the receptor's involvement in macrophage polarization.
Asunto(s)
Inflamación , Interleucina-4/farmacología , Proteínas Quinasas JNK Activadas por Mitógenos/fisiología , Activación de Macrófagos , Receptores Depuradores de Clase A/agonistas , Receptores Depuradores de Clase A/genética , Animales , Polaridad Celular/efectos de los fármacos , Polaridad Celular/genética , Células Cultivadas , Femenino , Humanos , Inflamación/inducido químicamente , Inflamación/genética , Inflamación/metabolismo , Mediadores de Inflamación/fisiología , Lipólisis/efectos de los fármacos , Lipólisis/genética , Lipoproteínas LDL/farmacología , Activación de Macrófagos/efectos de los fármacos , Activación de Macrófagos/genética , Macrófagos/efectos de los fármacos , Macrófagos/fisiología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Fagocitosis/efectos de los fármacos , Fagocitosis/genética , Polisacáridos/farmacología , Procesamiento Proteico-Postraduccional/genética , Células RAW 264.7 , Receptores Depuradores de Clase A/química , Receptores Depuradores de Clase A/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genética , Ubiquitinación/genéticaRESUMEN
Mucus alterations are a feature of ulcerative colitis (UC) and can drive inflammation by compromising the mucosal barrier to luminal bacteria. The exact pathogenesis of UC remains unclear, but CD4+ T cells reacting to commensal antigens appear to contribute to pathology. Given the unique capacity of dendritic cells (DCs) to activate naive T cells, colon DCs may activate pathogenic T cells and contribute to disease. Using Muc2-/- mice, which lack a functional mucus barrier and develop spontaneous colitis, we show that colitic animals have reduced colon CD103+ CD11b- DCs and increased CD103- CD11b+ phagocytes. Moreover, changes in colonic DC subsets and distinct cytokine patterns distinguish mice with distally localized colitis from mice with colitis spread proximally. Specifically, mice with proximally spread, but not distally contained, colitis have increased IL-1ß, IL-6, IL-17, TNFα, and IFNγ combined with decreased IL-10 in the distal colon. These individuals also have increased numbers of CD103+ CD11b+ DCs in the distal colon. CD103+ CD11b+ DCs isolated from colitic but not noncolitic mice induced robust differentiation of Th17 cells but not Th1 cells ex vivo. In contrast, CD103- CD11b+ DCs from colitic Muc2-/- mice induced Th17 as well as Th1 differentiation. Thus, the local environment influences the capacity of intestinal DC subsets to induce T cell proliferation and differentiation, with CD103+ CD11b+ DCs inducing IL-17-producing T cells being a key feature of extensively spread colitis.
Asunto(s)
Antígenos CD/metabolismo , Antígenos CD11/metabolismo , Colitis/inmunología , Colitis/patología , Células Dendríticas/inmunología , Cadenas alfa de Integrinas/metabolismo , Mucina 2/deficiencia , Células Th17/inmunología , Animales , Antígeno CD11b/metabolismo , Recuento de Células , Proliferación Celular , Colon/patología , Citocinas/metabolismo , Mediadores de Inflamación/metabolismo , Subgrupos Linfocitarios/inmunología , Ratones Endogámicos C57BL , Mucina 2/metabolismo , Neutrófilos/metabolismo , Recto/patologíaRESUMEN
The intestinal commensal microbiota is essential for many host physiological processes, but its impact on infectious diseases is poorly understood. Here we investigate the influence of the gut microbiota during oral Salmonella infection. We report a higher bacterial burden in mesenteric lymph nodes (MLN) of intragastrically infected germ-free (GF) mice compared to conventionally-raised (CONV-R) animals, despite similar inflammatory phagocyte recruitment. Salmonella penetration into the lamina propria of the small intestine and splenic bacterial burden were not altered in the absence of the microbiota. Intragastrically infected GF mice also displayed a higher frequency of IFN-γ-producing NK, NKT, CD4(+), and CD8(+) T cells in the MLN despite IL-12 levels similar to infected CONV-R mice. However, infecting mice intraperitoneally abrogated the difference in MLN bacterial load and IFN-γ-producing cells observed in intragastrically-infected animals. Moreover, mice treated with antibiotics (ABX) and intragastrically infected with Salmonella had a greater bacterial burden and frequency of IFN-γ-producing cells in the MLN. In ABX mice the number of Salmonella correlated with the frequency of IFN-γ-producing lymphocytes in the MLN, while no such correlation was observed in the MLN of infected GF mice. Overall, the data show that the lack of the microbiota influences pathogen colonization of the MLN, and the increased IFN-γ in the MLN of infected GF mice is not only due to the absence of commensals at the time of infection but the lack of immune signals provided by the microbiota from birth.
Asunto(s)
Antibiosis , Microbioma Gastrointestinal , Interferón gamma/metabolismo , Interleucina-12/metabolismo , Ganglios Linfáticos/inmunología , Ganglios Linfáticos/microbiología , Salmonelosis Animal/inmunología , Animales , Carga Bacteriana , Células Asesinas Naturales/inmunología , Ratones , Linfocitos T/inmunologíaRESUMEN
BACKGROUND: The colonic mucus layer plays a critical role in intestinal homeostasis by limiting contact between luminal bacteria and the mucosal immune system. A defective mucus barrier in animal models allows bacterial contact with the intestinal epithelium and results in spontaneous colitis. A defective mucus barrier is also a key feature of active ulcerative colitis (UC). Alterations in the immune compartment due to intestinal bacterial breach in mice lacking the colon mucus barrier have not been characterized and correlated to active UC. AIMS: To characterize alterations in the immune compartment due to intestinal bacterial breach in Muc2-/- mice, which lack the colon mucus barrier, and correlate the findings to active UC. METHODS: Bacterial contact with colon epithelium and penetration into colon tissue was examined in Muc2-/- mice and colon biopsies from patients with active UC using fluorescence microscopy and qPCR. Neutrophils, lymphocytes, CD103+ dendritic cell subsets and macrophages in colon from Muc2-/- mice and biopsies from UC patients were quantitated by flow cytometry. RESULTS: Inflamed UC patients and Muc2-/- mice had bacteria in contact with the colon epithelium. Bacterial rRNA was present in colonic mucosa in humans and Muc2-/- mice and in the draining lymph nodes of mice. Inflamed Muc2-/- mice and UC patients had elevated colon neutrophils, T cells and macrophages while a reduced frequency of CD103+ DCs was present in the inflamed colon of both mice and humans. CONCLUSIONS: The parallel features of the colon immune cell compartment in Muc2-/- mice and UC patients supports the usefulness of this model to understand the early phase of spontaneous colitis and will provide insight into novel strategies to treat UC.
Asunto(s)
Colitis Ulcerosa/patología , Mucina 2/deficiencia , Adulto , Anciano , Animales , Antígenos CD/metabolismo , Recuento de Células , Colitis Ulcerosa/microbiología , Colon/microbiología , Colon/patología , Células Dendríticas/patología , Femenino , Humanos , Inflamación/patología , Cadenas alfa de Integrinas/metabolismo , Mucosa Intestinal/patología , Receptores de Lipopolisacáridos/metabolismo , Macrófagos/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Persona de Mediana Edad , Mucina 2/metabolismo , Infiltración Neutrófila , Adulto JovenRESUMEN
Plasmacytoid dendritic cells (pDCs) are a subpopulation of dendritic cells specialized in the production of IFN-α/ß, particularly during viral infections. In this way pDCs directly impact antiviral immunity and influence T cell activation. However, despite their role as modulators of the immune response, their function as antigen-presenting cells (APCs) remains poorly understood. Indeed, their capacity as APCs during bacterial infections is unexplored. Here we investigate the importance of MyD88 and IFN-α/ß in upregulating costimulatory molecules on pDCs during Listeria infection and their impact on activation of naïve CD8 T cells. We show that pDCs efficiently upregulate CD80 and CD86 during systemic Listeria infection, yet express lower levels of these molecules than conventional dendritic cells (cDCs). Furthermore, pDCs are able to stimulate CD8 T cell proliferation and IFN-γ production, although less efficiently than cDCs. Despite these differences, the influence of MyD88 and IFN-α/ß on CD80 and CD86 expression on pDCs and cDCs is similar. Thus, our data show for the first time the potential of pDCs to activate CD8 T cells in response to a bacterial infection.