RESUMEN
Type I and type II IFNs are important immune modulators in both innate and adaptive immunity. They transmit signaling by activating JAK-STAT pathways. Sirtuin 1 (SIRT1), a class III NAD+-dependent deacetylase, has multiple functions in a variety of physiological processes. Here, we characterized the novel functions of SIRT1 in the regulation of type I and type II IFN-induced signaling. Overexpression of SIRT1 inhibited type I and type II IFN-induced interferon-stimulated response element activation. In contrast, knockout of SIRT1 promoted type I and type II IFN-induced expression of ISGs and inhibited viral replication. Treatment with SIRT1 inhibitor EX527 had similar positive effects. SIRT1 physically associated with STAT1 or STAT3, and this interaction was enhanced by IFN stimulation or viral infection. By deacetylating STAT1 at K673 and STAT3 at K679/K685/K707/K709, SIRT1 downregulated the phosphorylation of STAT1 (Y701) and STAT3 (Y705). Sirt1+/- primary peritoneal macrophages and Sirt1+/- mice exhibited enhanced IFN-induced signaling and antiviral activity. Thus, SIRT1 is a novel negative regulator of type I and type II IFN-induced signaling through its deacetylase activity.IMPORTANCESIRT1 has been reported in the precise regulation of antiviral (RNA and DNA) immunity. However, its functions in type I and type II IFN-induced signaling are still unclear. In this study, we deciphered the important functions of SIRT1 in both type I and type II IFN-induced JAK-STAT signaling and explored the potential acting mechanisms. It is helpful for understanding the regulatory roles of SIRT1 at different levels of IFN signaling. It also consolidates the notion that SIRT1 is an important target for intervention in viral infection, inflammatory diseases, or even interferon-related therapies.
Asunto(s)
Interferón Tipo I , Sirtuina 1 , Virosis , Animales , Ratones , Inmunidad Innata , Interferón Tipo I/metabolismo , Interferón gamma , Fosforilación , Transducción de Señal , Sirtuina 1/genética , Sirtuina 1/metabolismo , Factor de Transcripción STAT1/metabolismo , Virosis/inmunologíaRESUMEN
Ubiquitination and deubiquitination are important post-translational regulatory mechanisms responsible for fine tuning the antiviral signaling. In this study, we identified a deubiquitinase, the ubiquitin-specific peptidase 7/herpes virus associated ubiquitin-specific protease (USP7/HAUSP) as an important negative modulator of virus-induced signaling. Overexpression of USP7 suppressed Sendai virus and polyinosinic-polycytidylic acid and poly(deoxyadenylic-deoxythymidylic)-induced ISRE and IFN-ß activation, and enhanced virus replication. Knockdown or knockout of endogenous USP7 expression had the opposite effect. Coimmunoprecipitation assays showed that USP7 physically interacted with tripartite motif (TRIM)27. This interaction was enhanced after SeV infection. In addition, TNF receptor-associated factor family member-associated NF-kappa-B-binding kinase (TBK)-1 was pulled down in the TRIM27-USP7 complex. Overexpression of USP7 promoted the ubiquitination and degradation of TBK1 through promoting the stability of TRIM27. Knockout of endogenous USP7 led to enhanced TRIM27 degradation and reduced TBK1 ubiquitination and degradation, resulting in enhanced type I IFN signaling. Our findings suggest that USP7 acts as a negative regulator in antiviral signaling by stabilizing TRIM27 and promoting the degradation of TBK1.-Cai, J., Chen, H.-Y., Peng, S.-J., Meng, J.-L., Wang, Y., Zhou, Y., Qian, X.-P., Sun, X.-Y., Pang, X.-W., Zhang, Y., Zhang, J. USP7-TRIM27 axis negatively modulates antiviral type I IFN signaling.
Asunto(s)
Proteínas de Unión al ADN/metabolismo , Interferón Tipo I/metabolismo , Proteínas Nucleares/metabolismo , Infecciones por Respirovirus/metabolismo , Virus Sendai/metabolismo , Transducción de Señal , Peptidasa Específica de Ubiquitina 7/metabolismo , Proteínas de Unión al ADN/genética , Células HEK293 , Células HeLa , Humanos , Interferón Tipo I/genética , Proteínas Nucleares/genética , Proteolisis , Infecciones por Respirovirus/genética , Virus Sendai/genética , Peptidasa Específica de Ubiquitina 7/genética , UbiquitinaciónRESUMEN
Thymic involution happened early in life, but a certain ratio of activated CD4+ T cells will persistently recirculate into the thymus from the periphery and it have been suggested to be able to inhibit the development of embryonic thymocytes. Our present study was aimed to elucidate the specific mechanism how activated CD4+ T cells could influence upon developing thymocytes by using fetal thymic organ culture (FTOC) and kidney capsule transplantation. Our results demonstrated that Th2 cells were found to play a fundamental role in the inhibition of embryonic thymocyte development since a very low concentration of Th2 cells could obviously reduce the total number of thymocytes. And this effect was not tenable in other Th cell type. Notably, IL-4, the major cytokine secreted by Th2 cells, was suggested the key factor playing the inhibition role. In addition to reduced cell population, the proportion of double positive (DP) T cells was also heavily decreased. Furthermore, we demonstrated that it was the downstream effector signal transducer and activator of transcription 6 (STAT6) of IL-4 partially manipulate this inhibition. Together, these findings reveal a novel influence of Th2 cells re-entering the thymus on thymic involution.
Asunto(s)
Interleucina-4/metabolismo , Factor de Transcripción STAT6/metabolismo , Células Th2/inmunología , Timo/inmunología , Timo/fisiopatología , Animales , Diferenciación Celular , Técnicas de Cocultivo , Femenino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Técnicas de Cultivo de Órganos , Factor de Transcripción STAT6/deficiencia , Factor de Transcripción STAT6/genética , Transducción de Señal , Células Th2/patología , Timo/embriologíaRESUMEN
Thymic involution is evolutionarily conserved and occurs early in life. However, the physiological significance remains elusive of this seemingly detrimental process. The present study investigated the potential impact of altered thymic output on T cell memory using ovalbumin (OVA) expressed by Listeria monocytogenes as a model antigen. Suspension of thymic emigration by thymectomy was shown to lead to a marked increase in the frequency and total number of OVA-specific memory T cells. In contrast, oversupply of thymic emigrants through thymic grafting caused a significant decrease of such cells. When rechallenged with L. monocytogenes expressing OVA, the thymectomized mice mounted a more potent recall response as evidenced by the enlarged population of OVA-specific tetramer⺠cells and the accelerated clearance of the bacteria. Notably, the memory-enhancing effect of thymectomy was abrogated following weekly adoptive transfer of naive T cells. Together, data from the present study indicate that reduced thymic output favors the maintenance of the memory T cell pool.
Asunto(s)
Memoria Inmunológica , Listeria monocytogenes/inmunología , Listeriosis/inmunología , Ovalbúmina/inmunología , Linfocitos T/inmunología , Timo/citología , Traslado Adoptivo , Animales , Listeriosis/terapia , Ratones , Ratones Endogámicos C57BL , Linfocitos T/microbiología , Timectomía , Timo/inmunología , Timo/microbiologíaRESUMEN
Although the role of E proteins in the thymocyte development is well documented, much less is known about their function in peripheral T cells. Here we demonstrated that CD4 promoter-driven transgenic expression of Id1, a naturally occurring dominant-negative inhibitor of E proteins, can substitute for the co-stimulatory signal delivered by CD28 to facilitate the proliferation and survival of naïve CD4+ cells upon anti-CD3 stimulation. We next discovered that IL-2 production and NF-κB activity after anti-CD3 stimulation were significantly elevated in Id1-expressing cells, which may be, at least in part, responsible for the augmentation of their proliferation and survival. Taken together, results from this study suggest an important role of E and Id proteins in peripheral T cell activation. The ability of Id proteins to by-pass co-stimulatory signals to enable T cell activation has significant implications in regulating T cell immunity.
Asunto(s)
Linfocitos T CD4-Positivos/citología , Regulación de la Expresión Génica , Proteína 1 Inhibidora de la Diferenciación/biosíntesis , Receptores de Antígenos de Linfocitos T/metabolismo , Animales , Antígenos CD28/metabolismo , Complejo CD3/metabolismo , Proliferación Celular , Separación Celular , Supervivencia Celular , Relación Dosis-Respuesta a Droga , Citometría de Flujo , Interleucina-2/metabolismo , Activación de Linfocitos , Ratones , Ratones Transgénicos , FN-kappa B/metabolismoRESUMEN
Netrin-1, a known axon guidance molecule, being a secreted laminin-related molecule, has been suggested to be involved in multiple physiological and pathological conditions, such as organogenesis, angiogenesis, tumorigenesis, and inflammation-mediated tissue injury. However, its function in thymocyte development is still unknown. Here, we demonstrate that Netrin-1 is expressed in mouse thymus tissue and is primarily expressed in thymic stromal cells, and the expression of Netrin-1 in thymocytes can be induced by anti-CD3 antibody or IL-7 treatment. Importantly, Netrin-1 mediates the adhesion of thymocytes, and this effect is comparable to or greater than that of fibronectin. Furthermore, Netrin-1 specifically promotes the chemotaxis of CXCL12. These suggest that Netrin-1 may play an important role in thymocyte development.
Asunto(s)
Movimiento Celular/genética , Expresión Génica , Factores de Crecimiento Nervioso/genética , Timocitos/metabolismo , Timo/metabolismo , Proteínas Supresoras de Tumor/genética , Animales , Adhesión Celular/genética , Adhesión Celular/inmunología , Movimiento Celular/inmunología , Quimiocina CXCL12/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Interleucina-7/farmacología , Ratones , Factores de Crecimiento Nervioso/metabolismo , Receptores de Netrina , Netrina-1 , Receptores de Superficie Celular/genética , Receptores de Superficie Celular/metabolismo , Timocitos/inmunología , Timo/inmunología , Proteínas Supresoras de Tumor/metabolismoRESUMEN
The innate immune responses are tightly regulated to ensure effective clearance of invading pathogens and avoid excessive inflammation. Ubiquitination and deubiquitination are important post-translational modifications in antiviral immune responses. Here, we discovered deubiquitinase USP47 as a novel negative immune system regulator. Overexpression of USP47 repressed Sendai virus, poly(I:C) and poly(dA:dT)-induced ISRE and IFN-ß activation, along with reduced IFNB1 transcription and enhanced viral replication. Knockdown of USP47 expression had the opposite effects. Dual-luciferase and phosphorylation assays showed that USP47 targeted downstream of MAVS and upstream of TBK1. Additional co-immunoprecipitation assays suggested that USP47 interacted with TRAF3 and TRAF6. Importantly, USP47 removed K63-linked polyubiquitin chains from TRAF3 and TRAF6. Hence, we describe a novel modulator of the antiviral innate immune response, USP47, which removes K63-linked polyubiquitins from TRAF3 and TRAF6, leading to reduced type I IFN signaling.
Asunto(s)
Interferón Tipo I , Virus , Factor 3 Asociado a Receptor de TNF/genética , Factor 3 Asociado a Receptor de TNF/metabolismo , Factor 6 Asociado a Receptor de TNF/metabolismo , Inmunidad Innata , Interferón Tipo I/metabolismo , Antivirales , Ubiquitinación , Enzimas Desubicuitinizantes/metabolismoRESUMEN
Metastasis is the leading cause of cancer-related death. Despite the recent advancements in cancer treatment, there is currently no approved therapy for metastasis. The present study reveals a potent and selective activity of PRAK in the regulation of tumor metastasis. While showing no apparent effect on the growth of primary breast cancers or subcutaneously inoculated tumor lines, Prak deficiency abrogates lung metastases in PyMT mice or mice receiving intravenous injection of tumor cells. Consistently, PRAK expression is closely associated with metastatic risk in human cancers. Further analysis indicates that loss of function of PRAK leads to a pronounced inhibition of HIF-1α protein synthesis, possibly due to reduced mTORC1 activities. Notably, pharmacological inactivation of PRAK with a clinically relevant inhibitor recapitulates the anti-metastatic effect of Prak depletion, highlighting the therapeutic potential of targeting PRAK in the control of metastasis.
Asunto(s)
Péptidos y Proteínas de Señalización Intracelular/metabolismo , Metástasis de la Neoplasia , Neoplasias/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , Animales , Neoplasias de la Mama , Línea Celular Tumoral , Femenino , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Péptidos y Proteínas de Señalización Intracelular/genética , Neoplasias Pulmonares , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones SCID , Neoplasias/terapia , Proteínas Serina-Treonina Quinasas/genéticaRESUMEN
OBJECTIVE: To investigate the effect of tumor associate antigen of CHP2 on the metastatic potential of HEK293 cells in nude mice. METHODS: Matrigel testing by Boyden chamber and intraperitoneal inoculation of BALB/c nude mice were respectively performed to detect the metastatic potential of HEK293 transfectants in vitro and in vivo. HE staining was used to confirm the tumor metastasis in related organs. Molecules probably involved in metastasis of HEK293 cells were screened by RT-PCR. RESULTS: CHP2 significantly promoted the migration of HEK293 cells through matrigel in vitro. CHP2 accelerated the tumorigenesis of HEK293 cells in nude mice and tissue invasion was observed in spleen, kidney and liver. CHP2 up-regulated osteopontin (OPN) expression in HEK293 cells, but had no effect on MMP2 expression. CONCLUSION: CHP2 enhanced the migration and metastasis of HEK293 cells in vitro and in vivo. Up-regulated OPN expression might contribute to the invasive potential of HEK293 cells.
Asunto(s)
Proteínas de Unión al Calcio/genética , Neoplasias Hepáticas/secundario , Neoplasias Peritoneales/secundario , Neoplasias del Bazo/secundario , Animales , Células HEK293 , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Invasividad Neoplásica , Metástasis de la Neoplasia , Proteínas de Neoplasias/genética , Trasplante de Neoplasias , Neoplasias Peritoneales/patologíaRESUMEN
Parainfluenza virus infection is a common respiratory illness in children. Although lncRNAs are novel regulators of virus-induced innate immunity, a systemic attempt to characterize the differential expression of lncRNAs upon parainfluenza virus infection is lacking. In this report, we identify 207 lncRNAs and 166 mRNAs differentially expressed in SeV-infected HEK293T cells by microarray. The functional annotation analysis reveals that differentially regulated transcripts are predominantly involved in the host antiviral response pathway. The lncRNAs with the potential to regulate SeV-induced antiviral response are identified by building the lncRNA-mRNA coexpression network. Furthermore, silencing lncRNA ENST00000565297 results in reduced type I IFN signaling upon SeV infection. These catalogs may facilitate future analysis of the functions of lncRNAs in innate immunity and related diseases.
Asunto(s)
Inmunidad Innata/genética , Infecciones por Paramyxoviridae/genética , ARN Largo no Codificante/fisiología , Niño , Perfilación de la Expresión Génica , Células HEK293 , Humanos , Análisis de Secuencia por Matrices de Oligonucleótidos , Infecciones por Paramyxoviridae/inmunología , ARN Largo no Codificante/genética , Infecciones por Respirovirus/genética , Infecciones por Respirovirus/inmunología , Virus Sendai/inmunología , Virus Sendai/patogenicidad , TranscriptomaRESUMEN
Activated T cells have been shown to be able to recirculate into the thymus from the periphery. The present study was aimed to elucidate the functional consequences of thymic homing of activated T cells upon developing thymocytes and thymic epithelial cells (TEC). In the presence of activated T cells, especially CD4+ T cells, T cell development was found to be inhibited in thymic organ cultures with markedly reduced cellularity. Thymic transplantation demonstrated that the inhibitory effect was most likely due to a defective microenvironment. As the major component of the thymic stroma, the TEC compartment was severely disturbed after prolonged exposure to the activated T cells. In addition to reduced cell proliferation, TEC differentiation was heavily skewed to the mTEC lineage. Furthermore, we demonstrated that RANKL highly expressed by activated CD4+ T cells was primarily responsible for the detrimental effects. Presumably, excessive RANK signaling drove overproduction of mTECs and possibly exhaustion of epithelial progenitors, thereby facilitating the deterioration of the epithelial structures. These findings not only reveal a novel activity of activated T cells re-entering the thymus, but also provide a new perspective for understanding the mechanism underlying thymic involution.