RESUMEN
BACKGROUND: CD8+ T-lymphocyte subsets defined by killer lectin-like receptor G1 (KLRG1) and CD127 expression have been reported to have an important role in infection, but their role in the setting of lymphoid malignancies, specifically follicular lymphoma (FL), has not been studied. METHODS: To characterize the phenotype of KLRG1/CD127-defined CD8+ subsets, surface and intracellular markers were measured by flow cytometry and Cytometry by time of flight (CyTOF), and the transcriptional profile of these cells was determined by CITE-Seq (Cellular Indexing of Transcriptomes and Epitopes by Sequencing). The functional capacity of each subset was determined, as was their impact on overall survival (OS) and event-free survival (EFS) of patients with FL. RESULTS: We found that intratumoral CD8+ cells in FL are skewed toward effector cell subsets, particularly KLRG+CD127- and KLRG1-CD127- cells over memory cell subsets, such as KLRG1-CD127+ and KLRG1+CD127+ cells. While effector subsets exhibited increased capacity to produce cytokines/granules when compared with memory subsets, their proliferative capacity and viability were found to be substantially inferior. Clinically, a skewed distribution of intratumoral CD8+ T cells favoring effector subtypes was associated with an inferior outcome in patients with FL. Increased numbers of CD127+KLRG1- and CD127+KLRG1+ were significantly associated with a favorable OS and EFS, while CD127-KLRG1- correlated with a poor EFS and OS in patients with FL. Furthermore, we demonstrated that interleukin (IL)-15 promotes CD127-KLRG1+ cell development in the presence of dendritic cells via a phosphoinositide 3-kinase (PI3K)-dependent mechanism, and treatment of CD8+ T cells with a PI3K inhibitor downregulated the transcription factors responsible for CD127-KLRG1+ differentiation. CONCLUSIONS: Taken together, these results reveal not only a biological and prognostic role for KLRG1/CD127-defined CD8+ subsets in FL but also a potential role for PI3K inhibitors to manipulate the differentiation of CD8+ T cells, thereby promoting a more effective antitumor immune response.
Asunto(s)
Linfocitos T CD8-positivos/inmunología , Subunidad alfa del Receptor de Interleucina-7/metabolismo , Lectinas Tipo C/biosíntesis , Linfoma Folicular/metabolismo , Receptores Inmunológicos/biosíntesis , Diferenciación Celular/inmunología , Femenino , Humanos , Subunidad alfa del Receptor de Interleucina-7/biosíntesis , Subunidad alfa del Receptor de Interleucina-7/genética , Subunidad alfa del Receptor de Interleucina-7/inmunología , Lectinas Tipo C/genética , Lectinas Tipo C/inmunología , Linfoma Folicular/genética , Linfoma Folicular/inmunología , Receptores Inmunológicos/genética , Receptores Inmunológicos/inmunología , Resultado del Tratamiento , Microambiente TumoralRESUMEN
Neuromyelitis optica (NMO) is a chronic inflammatory demyelinating autoimmune disease of the central nervous system that most commonly affects the optic nerves and spinal cord. To characterize the immunological pathways involved in NMO, whole blood RNA expression array was performed using Nanostring nCounter technology. Two major clusters of genes were found associated with NMO: T cell-associated genes and the TNF/NF-kB signaling pathway. Analysis of the genes within the first cluster confirmed significantly reduced expression of IL7Ra (CD127) in the peripheral blood of NMO patients vs that in healthy controls. IL7Ra upstream transcription factors and its downstream survival signaling pathway were also markedly reduced. In line with the essential role of IL7Ra in T cell maturation and survival, a significantly lower number of naïve T cells, and reduced T cell survival signaling mediated by increased BID (BH3-interacting domain death agonist) expression and increased apoptosis was observed. Cumulatively, these findings indicate that the IL7Ra signaling pathway may play a role in the autoimmune process in NMO.
Asunto(s)
Subunidad alfa del Receptor de Interleucina-7/biosíntesis , Neuromielitis Óptica/metabolismo , Transducción de Señal/fisiología , Subgrupos de Linfocitos T/metabolismo , Adulto , Estudios de Cohortes , Femenino , Expresión Génica , Humanos , Subunidad alfa del Receptor de Interleucina-7/genética , Subunidad alfa del Receptor de Interleucina-7/inmunología , Masculino , Persona de Mediana Edad , Neuromielitis Óptica/genética , Neuromielitis Óptica/inmunología , Subgrupos de Linfocitos T/inmunologíaRESUMEN
Multiple sclerosis (MS) is a T cell-driven inflammatory disease of the CNS. Research on T cell subsets involved in MS pathogenesis has mainly focused on classical CD4+ T cells, especially Th17 cells, as they produce the proinflammatory, MS-associated cytokine IL-17. However, the abundant unconventional mucosal-associated invariant T (MAIT) cells are also able to produce IL-17. MAIT cells are characterized by high CD161 expression and a semi-invariant Vα7.2 TCR, with which they recognize bacterial and yeast Ags derived from the riboflavin (vitamin B2) metabolism. In this study, we characterized MAIT cells from the peripheral blood of MS patients in comparison with healthy individuals with respect to their type-17 differentiation. We found a specific increase of IL-17+ MAIT cells as well as an increased expression of retinoic acid-related orphan receptor (ROR)γt and CCR6 in MAIT cells from MS patients, whereas the expression of T cell activation markers HLA-DR and CD38 was not different. IL-17 production by MAIT cells furthermore correlated with the surface expression level of the IL-7 receptor α-chain (CD127), which was significantly increased on MAIT cells from MS patients in comparison with healthy individuals. In summary, our findings indicate an augmented type-17 differentiation of MAIT cells in MS patients associated with their IL-7 receptor surface expression, implicating a proinflammatory role of these unconventional T cells in MS immunopathology.
Asunto(s)
Sistema Nervioso Central/patología , Interleucina-17/biosíntesis , Subunidad alfa del Receptor de Interleucina-7/biosíntesis , Células T Invariantes Asociadas a Mucosa/inmunología , Esclerosis Múltiple/patología , ADP-Ribosil Ciclasa 1/metabolismo , Diferenciación Celular/inmunología , Células Cultivadas , Sistema Nervioso Central/inmunología , Antígenos HLA-DR/metabolismo , Humanos , Interferón gamma/biosíntesis , Activación de Linfocitos/inmunología , Glicoproteínas de Membrana/metabolismo , Células T Invariantes Asociadas a Mucosa/metabolismo , Esclerosis Múltiple/inmunología , Subfamilia B de Receptores Similares a Lectina de Células NK/biosíntesis , Miembro 3 del Grupo F de la Subfamilia 1 de Receptores Nucleares/biosíntesis , Receptores de Antígenos de Linfocitos T/inmunología , Receptores CCR6/biosíntesis , Factor de Transcripción STAT3/metabolismo , Factor de Transcripción STAT5/metabolismo , Transducción de Señal/inmunología , Proteínas Supresoras de Tumor/metabolismoRESUMEN
Acute lymphoblastic leukemia (ALL) remains the leading cause of cancer-related death in children and young adults. Compared to ALL in children, adult ALL has a much lower cure rate. Therefore, it is important to understand the molecular mechanisms underlying high-risk ALL and to develop therapeutic strategies that specifically target genes or pathways in ALL. Here, we explored the IL7R and SH2B3 expression in adult ALL and found that IL7R is significantly higher and Sh2B3 lower expressed in B-ALL compared to normal bone marrow control, and the IL7RhighSH2B3low is associated with high-risk factors, and with high relapse rate and low disease-free survival rate in the patients. We also found that Ikaros deletion was associated with the IL7RhighSH2B3low expression pattern and Ikaros directly binds the IL7R and SH2B3 promoter, and suppresses IL7R and promotes SH2B3 expression. On the other hand, casein kinase inhibitor, which increases Ikaros function, inhibits IL7R and stimulates SH2B3 expression in an Ikaros dependent manner. Our data indicate that IL7RhighSH2B3low expression distinguishes a novel subset of high-risk B-ALL associated with Ikaros dysfunction, and also suggest the therapeutic potential for treatment that combines casein kinase inhibitor, as an Ikaros activator, with drugs that target the IL7R signaling pathway.
Asunto(s)
Factor de Transcripción Ikaros/genética , Subunidad alfa del Receptor de Interleucina-7/biosíntesis , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Proteínas/metabolismo , Proteínas Adaptadoras Transductoras de Señales , Adulto , Biomarcadores de Tumor/análisis , Supervivencia sin Enfermedad , Femenino , Regulación Neoplásica de la Expresión Génica/genética , Humanos , Péptidos y Proteínas de Señalización Intracelular , Masculino , Leucemia-Linfoma Linfoblástico de Células Precursoras/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras/mortalidadRESUMEN
Generalized CD8+ T-cell impairment in chronic hepatitis C virus (HCV) infection and the contribution of liver-infiltrating CD8+ T-cells to the immunopathogenesis of this infection remain poorly understood. It is hypothesized that this impairment is partially due to reduced CD8+ T-cell activity in response to cytokines such as IL-7, particularly within the liver. To investigate this, the phenotype and cytokine responsiveness of blood- and liver-derived CD8+ T-cells from healthy controls and individuals with HCV infection were compared. In blood, IL-7 receptor α (CD127) expression on bulk CD8+ T-cells in HCV infection was no different than controls yet was lower on central memory T-cells, and there were fewer naïve cells. IL-7-induced signalling through phosphorylated STAT5 was lower in HCV infection than in controls, and differed between CD8+ T-cell subsets. Production of Bcl-2 following IL-7 stimulation was also lower in HCV infection and inversely related to the degree of liver fibrosis. In liver-derived CD8+ T-cells, STAT5 activation could not be increased with cytokine stimulation and basal Bcl-2 levels of liver-derived CD8+ T-cells were lower than blood-derived counterparts in HCV infection. Therefore, generalized CD8+ T-cell impairment in HCV infection is characterized, in part, by impaired IL-7-mediated signalling and survival, independent of CD127 expression. This impairment is more pronounced in the liver and may be associated with an increased potential for apoptosis. This generalized CD8+ T-cell impairment represents an important immune dysfunction in chronic HCV infection that may alter patient health.
Asunto(s)
Hepacivirus/efectos de los fármacos , Hepatitis C Crónica/inmunología , Subunidad alfa del Receptor de Interleucina-7/biosíntesis , Interleucina-7/metabolismo , Cirrosis Hepática/inmunología , Adulto , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/metabolismo , Linfocitos T CD8-positivos/virología , Femenino , Regulación de la Expresión Génica/inmunología , Hepacivirus/metabolismo , Hepatitis C Crónica/tratamiento farmacológico , Hepatitis C Crónica/patología , Hepatitis C Crónica/virología , Humanos , Interleucina-7/administración & dosificación , Subunidad alfa del Receptor de Interleucina-7/inmunología , Hígado/inmunología , Hígado/patología , Hígado/virología , Cirrosis Hepática/tratamiento farmacológico , Cirrosis Hepática/metabolismo , Cirrosis Hepática/virología , Masculino , Persona de Mediana Edad , Proteínas Proto-Oncogénicas c-bcl-2/biosíntesis , Factor de Transcripción STAT5RESUMEN
While the functional plasticity of memory CD4(+) T cells has been studied extensively, less is known about this property in memory CD8(+) T cells. Here, we report the direct measurement of plasticity by paired daughter analysis of effector and memory OT-I CD8(+) T cells primed in vivo with ovalbumin. Naïve, effector, and memory OT-I cells were isolated and activated in single-cell culture; then, after the first division, their daughter cells were transferred to new cultures with and without IL-4; expression of IFN-γ and IL-4 mRNAs was measured 5 days later in the resultant subclones. Approximately 40% of clonogenic memory CD8(+) T cells were bipotential in this assay, giving rise to an IL-4(-) subclone in the absence of IL-4 and an IL-4(+) subclone in the presence of IL-4. The frequency of bipotential cells was lower among memory cells than naïve cells but markedly higher than among 8-day effectors. Separation based on high or low expression of CD62L, CD122, CD127, or Ly6C did not identify a phenotypic marker of the bipotential cells. Functional plasticity in memory CD8(+) T-cell populations can therefore reflect modulation at the level of a single memory cell and its progeny.
Asunto(s)
Linfocitos T CD8-positivos/citología , Linfocitos T CD8-positivos/inmunología , Plasticidad de la Célula/inmunología , Memoria Inmunológica/inmunología , Interleucina-4/farmacología , Animales , Antígenos Ly/biosíntesis , Biomarcadores/análisis , Línea Celular , Interferón gamma/biosíntesis , Subunidad beta del Receptor de Interleucina-2/biosíntesis , Interleucina-4/genética , Subunidad alfa del Receptor de Interleucina-7/biosíntesis , Selectina L/biosíntesis , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , ARN Mensajero/biosíntesisRESUMEN
The IL-7R plays critical roles in lymphocyte development and homeostasis. Although IL-7R expression is strictly regulated during lymphocyte differentiation and the immune response, little is known regarding its in vivo regulation. To address this issue, we established a mouse line with targeted deletion of the conserved non-coding sequence 1 (CNS1) element found 3.6 kb upstream of the IL-7Rα promoter. We report that IL-7Rα is expressed normally on T and B cells in thymus and bone marrow of CNS1(-/-) mice except for in regulatory T cells. In contrast, these mice show reduced IL-7Rα expression in conventional CD4 and CD8 T cells as well as regulatory T, NKT, and γδ T cells in the periphery. CD4 T cells of CNS1(-/-) mice showed IL-7Rα upregulation in the absence of growth factors and IL-7Rα downregulation by IL-7 or TCR stimulation, although the expression levels were lower than those in control mice. Naive CD4 and CD8 T cells of CNS1(-/-) mice show attenuated survival by culture with IL-7 and reduced homeostatic proliferation after transfer into lymphopenic hosts. CNS1(-/-) mice exhibit impaired maintenance of Ag-stimulated T cells. Furthermore, IL-7Rα upregulation by glucocorticoids and TNF-α was abrogated in CNS1(-/-) mice. This work demonstrates that the CNS1 element controls IL-7Rα expression and maintenance of peripheral T cells, suggesting differential regulation of IL-7Rα expression between central and peripheral lymphoid organs.
Asunto(s)
Linfocitos T CD4-Positivos/citología , Linfocitos T CD8-positivos/citología , Elementos de Facilitación Genéticos , Subunidad alfa del Receptor de Interleucina-7/genética , Secuencias Reguladoras de Ácidos Nucleicos/genética , Animales , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Diferenciación Celular/inmunología , Proliferación Celular/genética , Células Cultivadas , Dexametasona/farmacología , Expresión Génica/efectos de los fármacos , Expresión Génica/genética , Subunidad alfa del Receptor de Interleucina-7/biosíntesis , Activación de Linfocitos/inmunología , Ratones , Ratones Noqueados , Regiones Promotoras Genéticas/genética , Eliminación de Secuencia/genética , Transducción de Señal/inmunología , Transcripción Genética/genética , Factor de Necrosis Tumoral alfa/farmacologíaRESUMEN
BACKGROUND: Zfat is a nuclear protein that harbours putative DNA-binding domains. T-cell specific deletion of Zfat in Zfat(f/f)-CD4Cre mice yields a significant decrease in the number of peripheral T-cells with a lower surface expression of interleukin-7 receptor-α (IL-7Rα). However, the molecular mechanism by which Zfat controls IL-7Rα expression remains unknown. MATERIALS AND METHODS: Expression levels of the molecules involved in IL-7Rα expression were determined by immunoblotting. RESULTS: Zfat-deficient peripheral T-cells showed a marked reduction in the FoxO1 protein that regulates IL-7Rα expression; however, the FoxO1 mRNA expression level was not affected by Zfat-deficiency. Furthermore, treatment of Zfat-deficient T-cells with a proteasome inhibitor, epoxomicin, restored FoxO1 expression levels, indicating that the loss of Zfat enhanced the proteasomal degradation of the FoxO1 protein. CONCLUSION: These results suggest that Zfat is required for peripheral T-cell homeostasis through IL-7Rα expression by controlling the FoxO1 protein.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Factores de Transcripción Forkhead/metabolismo , Complejo de la Endopetidasa Proteasomal/metabolismo , Factores de Transcripción/genética , Animales , Proteína Forkhead Box O1 , Factores de Transcripción Forkhead/biosíntesis , Factores de Transcripción Forkhead/genética , Subunidad alfa del Receptor de Interleucina-7/biosíntesis , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Oligopéptidos/farmacología , Inhibidores de Proteasoma/farmacología , ARN Mensajero/genéticaRESUMEN
The role of T-cell immunosenescence and functional CD8(+) T-cell responses in HIV/TB co-infection is unclear. We examined and correlated surrogate markers of HIV disease progression with immune activation, immunosenescence and differentiation using T-cell pools of HIV/TB co-infected, HIV-infected and healthy controls. Our investigations showed increased plasma viremia and reduced CD4/CD8 T-cell ratio in HIV/TB co-infected subjects relative to HIV-infected, and also a closer association with changes in the expression of CD38, a cyclic ADP ribose hydrolase and CD57, which were consistently expressed on late-senescent CD8(+) T cells. Up-regulation of CD57 and CD38 were directly proportional to lack of co-stimulatory markers on CD8(+) T cells, besides diminished expression of CD127 (IL-7Rα) on CD57(+)CD4(+) T cells. Notably, intracellular IFN-γ, perforin and granzyme B levels in HIV-specific CD8(+) T cells of HIV/TB co-infected subjects were diminished. Intracellular CD57 levels in HIV gag p24-specific CD8(+) T cells were significantly increased in HIV/TB co-infection. We suggest that HIV-TB co-infection contributes to senescence associated with chronic immune activation, which could be due to functional insufficiency of CD8(+) T cells.
Asunto(s)
Linfocitos T CD8-positivos/inmunología , Senescencia Celular/inmunología , Infecciones por VIH/inmunología , Inmunosenescencia/inmunología , Tuberculosis Pulmonar/inmunología , ADP-Ribosil Ciclasa 1/biosíntesis , Adulto , Relación CD4-CD8 , Antígenos CD57/biosíntesis , Antígenos CD57/metabolismo , Linfocitos T CD8-positivos/citología , Linfocitos T CD8-positivos/patología , Diferenciación Celular/inmunología , Proliferación Celular , Coinfección/inmunología , Progresión de la Enfermedad , Femenino , Granzimas/metabolismo , Cadenas alfa de HLA-DR/inmunología , Humanos , Interferón gamma/metabolismo , Subunidad alfa del Receptor de Interleucina-7/biosíntesis , Activación de Linfocitos/inmunología , Masculino , Glicoproteínas de Membrana/biosíntesis , Perforina/metabolismoRESUMEN
Adenosine A(2A) receptor (A(2A)R) blockade enhances innate and adaptive immune responses. However, mouse genetic studies have shown that A(2A)R deletion does not inhibit the growth of all tumor types. In the current study, we showed that growth rates for ectopic melanoma and bladder tumors are increased in Adora2a(-/-) mice within 2 weeks of tumor inoculation. A(2A)R deletion in the host reduced numbers of CD8(+) T cells and effector-memory differentiation of all T cells. To examine intrinsic functions in T cells, we generated mice harboring a T-cell-specific deletion of A(2A)R. In this host strain, tumor-bearing mice displayed increased growth of ectopic melanomas, decreased numbers of tumor-associated T cells, reduced effector-memory differentiation, and reduced antiapoptotic IL7Rα (CD127) expression on antigen-experienced cells. Intratumoral pharmacologic blockade similarly reduced CD8(+) T-cell density within tumors in wild-type hosts. We found that A(2A)R-proficient CD8(+) T cells specific for melanoma cells displayed a relative survival advantage in tumors. Thus, abrogating A(2A)R signaling appeared to reduce IL7R expression, survival, and differentiation of T cells in the tumor microenvironment. One implication of these results is that the antitumor effects of A(2A)R blockade that can be mediated by activation of cytotoxic T cells may be overcome in some tumor microenvironments as a result of impaired T-cell maintenance and effector-memory differentiation. Thus, our findings imply that the efficacious application of A(2A)R inhibitors for cancer immunotherapy may require careful dose optimization to prevent activation-induced T-cell death in tumors.
Asunto(s)
Antagonistas del Receptor de Adenosina A2/administración & dosificación , Linfocitos T CD8-positivos/efectos de los fármacos , Inmunoterapia , Receptor de Adenosina A2A/metabolismo , Inmunidad Adaptativa/efectos de los fármacos , Animales , Linfocitos T CD8-positivos/patología , Regulación Neoplásica de la Expresión Génica , Humanos , Inmunidad Innata/efectos de los fármacos , Subunidad alfa del Receptor de Interleucina-7/biosíntesis , Melanoma/tratamiento farmacológico , Melanoma/inmunología , Melanoma/patología , Ratones , Receptor de Adenosina A2A/genética , Microambiente Tumoral/efectos de los fármacos , Microambiente Tumoral/genética , Neoplasias de la Vejiga Urinaria/tratamiento farmacológico , Neoplasias de la Vejiga Urinaria/inmunología , Neoplasias de la Vejiga Urinaria/patologíaRESUMEN
Interleukin-7 is a non-redundant growth, differentiation and survival factor for human T lymphocytes. Most circulating, mature T cells express the receptor for IL-7, but not all. Importantly, CD4 Tregs express greatly reduced levels of IL-7R compared to conventional CD4 T cells, presenting an opportunity to selectively target the latter cells with either more IL-7 to boost responses, or to block IL-7 signalling to limit responses. This article reviews what is known about regulation of IL-7R expression, and recent progress in therapeutic approaches related to IL-7 and its receptor.
Asunto(s)
Subunidad alfa del Receptor de Interleucina-7/inmunología , Interleucina-7/inmunología , Interleucina-7/uso terapéutico , Activación de Linfocitos/inmunología , Linfocitos T Reguladores/inmunología , Humanos , Memoria Inmunológica/inmunología , Interleucina-7/biosíntesis , Subunidad alfa del Receptor de Interleucina-7/biosíntesis , Linfopenia/inmunología , Neoplasias/terapia , Transducción de Señal/inmunologíaRESUMEN
Expression of the IL-7 receptor α-chain (CD127) is decreased on CD8 T-cells in HIV infected patients and partially recovers in those receiving antiretroviral therapy with sustained viral suppression. We have shown that soluble HIV Tat protein down regulates CD127 expression on CD8 T-cells isolated from healthy HIV-negative individuals. Tat is taken up by CD8 T-cells via endocytosis, exits the endosome and then translocates to the inner leaflet of the cell membrane where it binds to the cytoplasmic tail of CD127 inducing receptor internalization and degradation by the proteasome. This down regulation of CD127 by Tat results in impaired CD8 T-cell function. Interestingly, suppression of CD127 by Tat is reversible and requires the continual presence of Tat in the culture media. We thus questioned whether the low IL-7 receptor expression evident on CD8 T-cells in HIV+ patients was similarly reversible and if suppression of the receptor could be maintained ex vivo by Tat protein alone. We show here that when CD8 T-cells isolated from HIV+ patients are incubated alone in fresh medium, low CD127 expression on the cell surface recovers to normal levels. This recovery of CD127, however, is completely inhibited by the addition of HIV Tat protein to the culture media. This study then provides evidence that soluble factor(s) are responsible for low CD127 expression on circulating CD8 T-cells in HIV+ individuals and further implicates Tat in suppressing this receptor essential to CD8 T-cell proliferation and function.
Asunto(s)
Linfocitos T CD8-positivos/inmunología , Infecciones por VIH/inmunología , Subunidad alfa del Receptor de Interleucina-7/biosíntesis , Productos del Gen tat del Virus de la Inmunodeficiencia Humana/farmacología , Adulto , Fármacos Anti-VIH/uso terapéutico , Terapia Antirretroviral Altamente Activa , Relación CD4-CD8 , Proliferación Celular , Infecciones por VIH/tratamiento farmacológico , Humanos , Subunidad alfa del Receptor de Interleucina-7/inmunología , Activación de Linfocitos/inmunología , Masculino , Persona de Mediana Edad , Transducción de Señal/inmunología , Adulto Joven , Productos del Gen tat del Virus de la Inmunodeficiencia Humana/metabolismoRESUMEN
During infection or vaccination, only a small proportion of CD8(+) T cells differentiate into memory cells. The mechanisms underlying the differentiation of CD8(+) T cells into short-lived effector cells (SLECs) or memory precursor effector cells are poorly defined. It was recently shown in infectious models that the transcriptional repressor B lymphocyte-induced maturation protein 1 (Blimp-1) enhances the formation of SLECs. The factors controlling Blimp-1 expression leading to the in vivo formation of SLECs are still not known. However, it has been shown that cytokines such as IL-2 induce Blimp-1 expression in vitro. In this study, we took advantage of the low-inflammation model of dendritic cell immunization to study the role of the IL-2/Blimp-1 axis in SLEC differentiation as well as the importance of Blimp-1 expression in memory precursor effector cells for proper CD8(+) memory generation. Our results show that Blimp-1 deficiency affects effector differentiation and function in the absence of inflammation. Unexpectedly, memory generation was not affected in Blimp-1-deficient OT-I cells responding to vaccination. In addition, modulation of the bioavailability of IL-2 by injection either of a blocking Ab or of the cytokine, demonstrates a link between IL-2, Blimp-1 induction, and SLEC formation in wild-type cells. Conversely, injection of IL-2 had less effect on Blimp-1-deficient CD8(+) T cells, indicating that the effect of IL-2 on in vivo SLEC differentiation is mediated by Blimp-1. In conclusion, IL-2 induction of Blimp-1 expression is a key regulator of SLEC differentiation in vivo.
Asunto(s)
Linfocitos T CD8-positivos/inmunología , Vacunas contra el Cáncer/inmunología , Memoria Inmunológica , Interleucina-2/inmunología , Factores de Transcripción/biosíntesis , Animales , Anticuerpos Bloqueadores/inmunología , Apoptosis/inmunología , Diferenciación Celular/inmunología , Células Cultivadas , Células Dendríticas/inmunología , Femenino , Granzimas/biosíntesis , Inflamación/inmunología , Interferón gamma/biosíntesis , Interleucina-2/biosíntesis , Subunidad alfa del Receptor de Interleucina-7/biosíntesis , Subunidad alfa del Receptor de Interleucina-7/genética , Lectinas Tipo C , Listeria monocytogenes/genética , Activación de Linfocitos/inmunología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Ovalbúmina/inmunología , Factor 1 de Unión al Dominio 1 de Regulación Positiva , Receptores Inmunológicos/biosíntesis , Receptores Inmunológicos/genética , Factores de Transcripción/genéticaRESUMEN
BACKGROUND: During HIV infection distinct mechanisms drive immune activation of the CD4 and CD8 T cells leading to CD4 T-cell depletion and expansion of the CD8 T-cell pool. This immune activation is polyclonal and extends beyond HIV-specific T cells. One consequence of this immune activation is a profound decrease in IL-7Rα (CD127) expression on memory CD8 T cells. The mechanisms leading to this are unknown and because of the potential impact of reduced IL-7 signaling in memory T cells specific to HIV and other pathogens, in the present study we examined the molecular mechanisms implicated in this downregulation of CD127. METHODS: Membrane bound (mIL7RA) and soluble (sIL7RA) mRNA expression was determined by qRT-PCR. CD127, Eomesodermin (Eomes) and T-bet expression in healthy controls and HIV-infected patients were studied by flow cytometry. RESULTS: CD127 downregulation occurs at the transcriptional level for both mIL7RA and sIL7RA alternative spliced forms in the CD127 memory CD8 T cells. CD127 memory CD8 T cells exhibited increased Eomes expression and an 'effector-like' gene profile. These changes were associated with higher HIV-RNA levels. Following combination antiretroviral therapy (cART), there was an increase in CD127 expression over an extended period of time (>5 months) which was associated with decreased Eomes expression. CONCLUSION: CD127 is downregulated at a transcriptional level in memory CD8 T cells in association with upregulation of Eomes expression.
Asunto(s)
Linfocitos T CD8-positivos/inmunología , Infecciones por VIH/inmunología , Subunidad alfa del Receptor de Interleucina-7/biosíntesis , Proteínas de Dominio T Box/biosíntesis , Estudios de Cohortes , Estudios Transversales , Citometría de Flujo , Expresión Génica , Perfilación de la Expresión Génica , Humanos , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
Treg cells are critical for the prevention of autoimmune diseases and are thus prime candidates for cell-based clinical therapy. However, human Treg cells are "plastic", and are able to produce IL-17 under inflammatory conditions. Here, we identify and characterize the human Treg subpopulation that can be induced to produce IL-17 and identify its mechanisms. We confirm that a subpopulation of human Treg cells produces IL-17 in vitro when activated in the presence of IL-1ß, but not IL-6. "IL-17 potential" is restricted to population III (CD4(+) CD25(hi) CD127(lo) CD45RA(-) ) Treg cells expressing the natural killer cell marker CD161. We show that these cells are functionally as suppressive and have similar phenotypic/molecular characteristics to other subpopulations of Treg cells and retain their suppressive function following IL-17 induction. Importantly, we find that IL-17 production is STAT3 dependent, with Treg cells from patients with STAT3 mutations unable to make IL-17. Finally, we show that CD161(+) population III Treg cells accumulate in inflamed joints of patients with inflammatory arthritis and are the predominant IL-17-producing Treg-cell population at these sites. As IL-17 production from this Treg-cell subpopulation is not accompanied by a loss of regulatory function, in the context of cell therapy, exclusion of these cells from the cell product may not be necessary.
Asunto(s)
Interleucina-17/biosíntesis , Subfamilia B de Receptores Similares a Lectina de Células NK/metabolismo , Factor de Transcripción STAT3/metabolismo , Linfocitos T Reguladores/inmunología , Linfocitos T Reguladores/metabolismo , Adulto , Anciano , Antígenos CD4/biosíntesis , Células Cultivadas , Femenino , Factores de Transcripción Forkhead/biosíntesis , Humanos , Interleucina-1beta/metabolismo , Subunidad alfa del Receptor de Interleucina-2/biosíntesis , Interleucina-6/metabolismo , Subunidad alfa del Receptor de Interleucina-7/biosíntesis , Antígenos Comunes de Leucocito/biosíntesis , Activación de Linfocitos , Masculino , Persona de Mediana Edad , Subfamilia B de Receptores Similares a Lectina de Células NK/inmunología , Factor de Transcripción STAT3/genéticaRESUMEN
Regulatory T (Treg) cells suppress autoimmune disease, and impaired Treg cell function is associated with rheumatoid arthritis. Here we demonstrate that forkhead box P3 (FOXP3) transcriptional activity and, consequently, Treg cell suppressive function are regulated by phosphorylation at Ser418 in the C-terminal DNA-binding domain. In rheumatoid arthritis-derived Treg cells, the Ser418 site was specifically dephosphorylated by protein phosphatase 1 (PP1), whose expression and enzymatic activity were induced in the inflamed synovium by tumor necrosis factor α (TNF-α), leading to impaired Treg cell function. Moreover, TNF-α-induced Treg cell dysfunction correlated with increased numbers of interleukin-17 (IL-17)(+) and interferon-γ (IFN-γ)(+)CD4(+) T cells within the inflamed synovium in rheumatoid arthritis. Treatment with a TNF-α-specific antibody restored Treg cell function in subjects with rheumatoid arthritis, which was associated with decreased PP1 expression and increased FOXP3 phosphorylation in Treg cells. Thus, TNF-α controls the balance between Treg cells and pathogenic TH17 and TH1 cells in the synovium of individuals with rheumatoid arthritis through FOXP3 dephosphorylation.
Asunto(s)
Artritis Reumatoide/inmunología , Artritis Reumatoide/metabolismo , Factores de Transcripción Forkhead/metabolismo , Linfocitos T Reguladores/inmunología , Factor de Necrosis Tumoral alfa/metabolismo , Células Cultivadas , Proteínas de Unión al ADN/metabolismo , Factores de Transcripción Forkhead/química , Humanos , Interferón gamma/biosíntesis , Interleucina-17/biosíntesis , Subunidad alfa del Receptor de Interleucina-2/biosíntesis , Subunidad alfa del Receptor de Interleucina-7/biosíntesis , Fosforilación , Proteína Fosfatasa 1/metabolismo , Interferencia de ARN , ARN Interferente Pequeño , Membrana Sinovial/inmunología , Membrana Sinovial/metabolismo , Linfocitos T Reguladores/metabolismo , Células TH1/inmunología , Células TH1/metabolismo , Células Th17/inmunología , Células Th17/metabolismo , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
In a previous study, we identified TRIB1, a serine-threonine kinase-like molecule, as a biomarker of chronic antibody-mediated rejection of human kidneys when measured in peripheral blood mononuclear cells. Here, we focused our analysis on a specific subset of peripheral blood mononuclear cells that play a dominant role in regulating immune responses in health and disease, so-called CD4(+)CD25(+)Foxp3(+) regulatory T cells (Tregs). We isolated both human and murine Treg and non-Treg counterparts and analyzed TRIB1 and Foxp3 mRNA expression by quantitative PCR on the freshly isolated cells or following 24 h of activation. Physical interaction between the human TRIB1 and Foxp3 proteins was analyzed in live cell lines by protein complementation assay using both flow cytometry and microscopy and confirmed in primary freshly isolated human CD4(+)CD25(hi)CD127(-) Tregs by co-immunoprecipitation. Both TRIB1 and Foxp3 were expressed at significantly higher levels in Tregs than in their CD4(+)CD25(-) counterparts (p < 0.001). Moreover, TRIB1 and Foxp3 mRNA levels correlated tightly in Tregs (Spearman r = 1.0; p < 0.001, n = 7), but not in CD4(+)CD25(-) T cells. The protein complementation assay revealed a direct physical interaction between TRIB1 and Foxp3 in live cells. This interaction was impaired upon deletion of the TRIB1 N-terminal but not the C-terminal domain, suggesting an interaction in the nucleus. This direct interaction within the nucleus was confirmed in primary human Tregs by co-immunoprecipitation. These data show a direct relationship between TRIB1 and Foxp3 in terms of their expression and physical interaction and highlight Tribbles-1 as a novel binding partner of Foxp3 in Tregs.
Asunto(s)
Factores de Transcripción Forkhead/metabolismo , Péptidos y Proteínas de Señalización Intracelular/química , Proteínas Serina-Treonina Quinasas/antagonistas & inhibidores , Linfocitos T Reguladores/citología , Animales , Biomarcadores/metabolismo , Linfocitos T CD4-Positivos/inmunología , Adhesión Celular , Ciclo Celular , Proliferación Celular , Supervivencia Celular , ADN Complementario/metabolismo , Prueba de Complementación Genética , Células HEK293 , Humanos , Subunidad alfa del Receptor de Interleucina-2/biosíntesis , Subunidad alfa del Receptor de Interleucina-7/biosíntesis , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Leucocitos Mononucleares/citología , Ratones , Unión Proteica , Proteínas Serina-Treonina Quinasas/química , Proteínas Serina-Treonina Quinasas/metabolismo , ARN Mensajero/metabolismo , Linfocitos T/citología , Linfocitos T Reguladores/metabolismoRESUMEN
CD8(+) T cells play a crucial role in the clearance of intracellular pathogens through the generation of cytotoxic effector cells that eliminate infected cells and long-lived memory cells that provide enhanced protection against reinfection. We have previously shown that the inhibitor of E protein transcription factors, Id2, is necessary for accumulation of effector and memory CD8(+) T cells during infection. In this study, we show that CD8(+) T cells lacking Id2 did not generate a robust terminally differentiated killer cell lectin-like receptor G1 (KLRG1)(hi) effector population, but displayed a cell-surface phenotype and cytokine profile consistent with memory precursors, raising the question as to whether loss of Id2 impairs the differentiation and/or survival of effector memory cells. We found that deletion of Bim rescued Id2-deficient CD8(+) cell survival during infection. However, the dramatic reduction in KLRG1(hi) cells caused by loss of Id2 remained in the absence of Bim, such that Id2/Bim double-deficient cells form an exclusively KLRG1(lo)CD127(hi) memory precursor population. Thus, we describe a role for Id2 in both the survival and differentiation of normal CD8(+) effector and memory populations.
Asunto(s)
Linfocitos T CD8-positivos/inmunología , Proteína 2 Inhibidora de la Diferenciación/fisiología , Receptores Inmunológicos/biosíntesis , Traslado Adoptivo , Animales , Linfocitos T CD8-positivos/microbiología , Linfocitos T CD8-positivos/virología , Supervivencia Celular/genética , Supervivencia Celular/inmunología , Citocinas/biosíntesis , Memoria Inmunológica/genética , Inmunofenotipificación , Proteína 2 Inhibidora de la Diferenciación/deficiencia , Proteína 2 Inhibidora de la Diferenciación/genética , Subunidad alfa del Receptor de Interleucina-7/biosíntesis , Lectinas Tipo C , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones Transgénicos , Células Madre/inmunología , Células Madre/microbiología , Células Madre/virología , Proteína bcl-X/deficiencia , Proteína bcl-X/genéticaRESUMEN
BACKGROUND & AIMS: Dysfunctional CD8(+) T cells are believed to contribute to the ability of hepatitis C virus (HCV) to evade the immune response. Most studies have focused on the effector functions of HCV-specific CD8(+) T cells or their surface expression of inhibitory receptors. There is currently no information available about the ex vivo ability of HCV-specific CD8(+) T cells to inhibit viral replication (antiviral efficacy). METHODS: To analyze the antiviral efficacy of virus-specific CD8(+) T cells ex vivo, we used an immunologic model based on a cell line that expresses HLA-A*02 and contains a stably replicating HCV reporter replicon. We isolated HCV-specific CD8(+) T cells from 18 HLA-A*02-positive patients with chronic HCV infection and 15 subjects with resolved HCV infection (7 spontaneous, 8 after therapy). Replicon cells were labeled with virus-specific peptides; inhibition of HCV replication was determined by measuring luciferase activity after 72 hours of coculture with virus-specific CD8(+) T cells. RESULTS: HCV-specific CD8(+) T cells from patients with chronic HCV infection had a significantly lower antiviral efficacy than influenza-, Epstein-Barr virus-, and cytomegalovirus-specific CD8(+) T cells. Antiviral efficacy was associated with the ability of virus-specific CD8(+) T cells to secrete interferon gamma. The antiviral efficacy of HCV-specific CD8(+) T cells was linked to surface expression of CD127 and PD-1. The cytokines interleukin-2, interleukin-7, and interleukin-15 increased the antiviral efficacy of CD127-positive but not of CD127-negative, HCV-specific CD8(+) T cells. Spontaneous, but not antiviral therapy-induced, viral clearance was associated with increased antiviral efficacy. CONCLUSIONS: The ability of CD8(+) T cells to inhibit HCV replication ex vivo is associated with their ability to secrete interferon gamma and their surface expression of CD127 and PD-1.