RESUMEN
BACKGROUND: Dysregulation of immune surveillance is tightly linked to the development of metabolic dysfunction-associated steatohepatitis (MASH)-driven hepatocellular carcinoma (HCC); however, its underlying mechanisms remain unclear. Herein, we aimed to determine the role of interleukin-21 receptor (IL-21R) in MASH-driven HCC. METHODS: The clinical significance of IL-21R was assessed in human HCC specimens using immunohistochemistry staining. Furthermore, the expression of IL-21R in mice was assessed in the STAM model. Thereafter, two different MASH-driven HCC mouse models were applied between IL-21R-deficient mice and wild type controls to explore the role of IL-21R in MASH-driven HCC. To further elucidate the potential mechanisms by which IL-21R affected MASH-driven HCC, whole transcriptome sequencing, flow cytometry and adoptive lymphocyte transfer were performed. Finally, flow cytometry, enzyme-linked immunosorbent assay, immunofluorescent staining, chromatin immunoprecipitation assay and western blotting were conducted to explore the mechanism by which IL-21R induced IgA+ B cells. RESULTS: HCC patients with high IL-21R expression exhibited poor relapse-free survival, advanced TNM stage and severe steatosis. Additionally, IL-21R was demonstrated to be upregulated in mouse liver tumors. Particularly, ablation of IL-21R impeded MASH-driven hepatocarcinogenesis with dramatically reduction of lipid accumulation. Moreover, cytotoxic CD8+ T lymphocyte activation was enhanced in the absence of IL-21R due to the reduction of immunosuppressive IgA+ B cells. Mechanistically, the IL-21R-STAT1-c-Jun/c-Fos regulatory axis was activated in MASH-driven HCC and thus promoted the transcription of Igha, resulting in the induction of IgA+ B cells. CONCLUSIONS: IL-21R plays a cancer-promoting role by inducing IgA+ B cells in MASH-driven hepatocarcinogenesis. Targeting IL-21R signaling represents a potential therapeutic strategy for cancer therapy.
Asunto(s)
Linfocitos B , Carcinoma Hepatocelular , Hígado Graso , Inmunoglobulina A , Neoplasias Hepáticas , Transducción de Señal , Animales , Humanos , Masculino , Ratones , Linfocitos B/metabolismo , Linfocitos B/inmunología , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patología , Carcinoma Hepatocelular/etiología , Carcinoma Hepatocelular/inmunología , Carcinoma Hepatocelular/genética , Línea Celular Tumoral , Modelos Animales de Enfermedad , Hígado Graso/metabolismo , Hígado Graso/patología , Hígado Graso/etiología , Regulación Neoplásica de la Expresión Génica , Inmunoglobulina A/metabolismo , Subunidad alfa del Receptor de Interleucina-21/metabolismo , Subunidad alfa del Receptor de Interleucina-21/genética , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patología , Neoplasias Hepáticas/etiología , Neoplasias Hepáticas/inmunología , Neoplasias Hepáticas/genética , Receptores de Interleucina-21/metabolismo , Receptores de Interleucina-21/genéticaRESUMEN
BACKGROUND: We identified a homologue of IL-21R (LcIL-21R) in large yellow croaker (Larimichthys crocea, Lc). Our investigation focused on understanding the molecular structural features and immune function of LcIL-21R. METHODS: We cloned the LcIL-21R gene from the genome of Larimichthys crocea by RTâPCR, and the molecular and structural characteristics of LcIL-21R were analyzed by a series of protein analysis tools. We used real-time PCR to investigate the tissue distribution of LcIL-21R, and LcIL-21R gene expression regulation was also measured in head kidney leukocytes under trivalent bacterial vaccine or poly (I:C) stimulation. RESULTS: The open reading frame (ORF) of the LcIL-21R gene is 1629 bp long and encodes a precursor protein of 542 amino acids (aa), with a 23-aa signal peptide and a 519-aa mature peptide containing four putative N-glycosylation sites. LcIL-21R has two fibronectin type III (FNIII)-like domains (D1 and D2), a transmembrane domain, and a cytoplasmic region. A conserved WSXWS motif was also found in the D2 domain. The predicted structure of the extracellular region of LcIL-21R (LcIL-21R-Ex) is highly similar to that of human IL-21R. LcIL-21R was constitutively expressed in all tissues examined, and LcIL-21R mRNA levels were increased in the head kidney and spleen upon inactivated trivalent bacterial vaccine or poly(I:C) stimulation. CONCLUSIONS: Our results suggest that LcIL-21R shares structural and functional properties with IL-21Rs found in other vertebrates, indicating its potential involvement in the IL-21-mediated immune response to pathogenic infections. These findings contribute to our understanding of the evolutionary conservation of IL-21 signaling and its role in the immune system.
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Perciformes , Receptores de Interleucina-21 , Animales , Humanos , Receptores de Interleucina-21/genética , Receptores de Interleucina-21/metabolismo , Secuencia de Aminoácidos , Regulación de la Expresión Génica , Perciformes/metabolismo , Vacunas Bacterianas , Proteínas de Peces/metabolismo , FilogeniaRESUMEN
Interleukin-21 and its receptors (IL-21/IL-21R) aggravate chlamydial lung infection, while macrophages (Mφ) are one of the main cells infected by chlamydia and the main source of inflammatory cytokines. Therefore, it is particularly important to study whether IL-21/IL-21R aggravates chlamydia respiratory infection by regulating Mφ. Combined with bioinformatics analysis, we established an IL-21R-deficient (IL-21R-/-) mouse model of Chlamydia muridarum (C. muridarum) respiratory tract infection in vivo, studied C. muridarum-stimulated RAW264.7 by the addition of rmIL-21 in vitro, and conducted adoptive transfer experiments to clarify the association between IL-21/IL-21R and Mφ. IL-21R-/- mice showed lower infiltration of pulmonary total Mφ, alveolar macrophages, and interstitial macrophages compared with WT mice following infection. Transcriptomic analysis suggested that M1-related genes are downregulated in IL-21R-/- mice and that IL-21R deficiency affects the Mφ-mediated inflammatory response during C. muridarum infection. In vivo experiments verified that in IL-21R-/- mice, pulmonary M1-type CD80+, CD86+, MHC II+, TNFα+, and iNOS+ Mφ decreased, while there were no differences in M2-type CD206+, TGF-ß+, IL-10+ and ARG1+ Mφ. In vitro, administration of rmIL-21 to C. muridarum-stimulated RAW264.7 cells promoted the levels of iNOS-NO and the expression of IL-12p40 and TNFα, but had no effect on TGFß or IL-10. Further, adoptive transfer of M1-like bone marrow-derived macrophages derived from IL-21R-/- mice, unlike those from WT mice, effectively protected the recipients against C. muridarum infection and induced relieved pulmonary pathology. These findings help in understanding the mechanism by which IL-21/IL-21R exacerbates chlamydia respiratory infection by promoting the proinflammatory effect of Mφ.
Asunto(s)
Infecciones por Chlamydia , Chlamydia muridarum , Animales , Ratones , Interleucina-10 , Factor de Necrosis Tumoral alfa , MacrófagosRESUMEN
Biallelic inactivating mutations in IL21R causes a combined immunodeficiency that is often complicated by cryptosporidium infections. While eight IL-21R-deficient patients have been reported previously, the natural course, immune characteristics of disease, and response to hematopoietic stem cell transplantation (HSCT) remain to be comprehensively examined. In our study, we have collected clinical histories of 13 patients with IL-21R deficiency from eight families across seven centers worldwide, including five novel patients identified by exome or NGS panel sequencing. Eight unique mutations in IL21R were identified in these patients, including two novel mutations. Median age at disease onset was 2.5 years (0.5-7 years). The main clinical manifestations were recurrent bacterial (84.6%), fungal (46.2%), and viral (38.5%) infections; cryptosporidiosis-associated cholangitis (46.2%); and asthma (23.1%). Inflammatory skin diseases (15.3%) and recurrent anaphylaxis (7.9%) constitute novel phenotypes of this combined immunodeficiency. Most patients exhibited hypogammaglobulinemia and reduced proportions of memory B cells, circulating T follicular helper cells, MAIT cells and terminally differentiated NK cells. However, IgE levels were elevated in 50% of IL-21R-deficient patients. Overall survival following HSCT (6 patients, mean follow-up 1.8 year) was 33.3%, with pre-existing organ damage constituting a negative prognostic factor. Mortality of non-transplanted patients (n = 7) was 57.1%. Our detailed analysis of the largest cohort of IL-21R-deficient patients to date provides in-depth clinical, immunological and immunophenotypic features of these patients, thereby establishing critical non-redundant functions of IL-21/IL-21R signaling in lymphocyte differentiation, humoral immunity and host defense against infection, and mechanisms of disease pathogenesis due to IL-21R deficiency. Outcome following HSCT depends on prior chronic infections and organ damage, which should thus be considered as early as possible following molecular diagnosis.
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Subunidad alfa del Receptor de Interleucina-21/deficiencia , Subunidad alfa del Receptor de Interleucina-21/genética , Adolescente , Linfocitos B/inmunología , Diferenciación Celular/genética , Diferenciación Celular/inmunología , Niño , Preescolar , Criptosporidiosis/genética , Criptosporidiosis/inmunología , Cryptosporidium/inmunología , Femenino , Genómica/métodos , Humanos , Inmunidad Humoral/genética , Inmunidad Humoral/inmunología , Lactante , Subunidad alfa del Receptor de Interleucina-21/inmunología , Activación de Linfocitos/genética , Activación de Linfocitos/inmunología , Masculino , Células B de Memoria/inmunología , Infección Persistente/genética , Infección Persistente/inmunología , Fenotipo , Transducción de Señal/genética , Transducción de Señal/inmunología , Adulto JovenRESUMEN
Respiratory syncytial virus (RSV) is the major cause of lower respiratory tract infections in children under 1 year. RSV vaccines are currently unavailable, and children suffering from multiple reinfections by the same viral strain fail to develop protective responses. Although RSV-specific antibodies can be detected upon infection, these have limited neutralizing capacity. Follicular helper T (Tfh) cells are specialized in providing signals to B cells and help the production and affinity maturation of antibodies, mainly via interleukin (IL) 21 secretion. In this study, we evaluated whether RSV could inhibit Tfh responses. We observed that Tfh cells fail to upregulate IL-21 production upon RSV infection. In the lungs, RSV infection downregulated the expression of IL-21/interleukin-21 receptor (IL-21R) in Tfh cells and upregulated programmed death-ligand 1 (PD-L1) expression in dendritic cells (DCs) and B cells. PD-L1 blockade during infection recovered IL-21R expression in Tfh cells and increased the secretion of IL-21 in a DC-dependent manner. IL-21 treatment decreased RSV viral load and lung inflammation, inducing the formation of tertiary lymphoid organs in the lung. It also decreased regulatory follicular T cells, and increased Tfh cells, B cells, antibody avidity and neutralization capacity, leading to an overall improved anti-RSV humoral response in infected mice. Passive immunization with purified immunoglobulin G from IL-21-treated RSV-infected mice protected against RSV infection. Our results unveil a pathway by which RSV affects Tfh cells by increasing PD-L1 expression on antigen-presenting cells, highlighting the importance of an IL-21-PD-L1 axis for the generation of protective responses to RSV infection.
Asunto(s)
Anticuerpos Neutralizantes , Infecciones por Virus Sincitial Respiratorio , Animales , Anticuerpos Antivirales , Interleucinas , Ratones , Infecciones por Virus Sincitial Respiratorio/terapia , Células T Auxiliares FolicularesRESUMEN
In our previous study, we explored the therapeutic effect of berberine (BBR) against IL-21/IL-21R mediated inflammatory proliferation of adjuvant-induced arthritic fibroblast-like synoviocytes (AA-FLS) through the PI3K/Akt pathway. The current study was designed to explore the therapeutic potential of BBR (15-45 µM) against IL-21/IL-21R mediated autophagy in AA-FLS mediated through PI3K/Akt signaling and Th17/Treg imbalance. Upon IL-21 stimulation, AA-FLS expressed elevated levels of autophagy-related 5 (Atg5), Beclin-1 and LC3-phosphatidylethanolamine conjugate 3-II (LC3-II) through the utilization of p62 and inhibition of C/EBP homologous protein (CHOP). BBR (15-45 µM) inhibited autophagy in AA-FLS cells mediated through PI3K/Akt signaling via suppressing autophagic elements, p62 sequestration and induction of CHOP in a dose-dependent manner. Moreover, IL-21 promoted the uncontrolled proliferation of AA-FLS through induction of B cell lymphoma-2 (Bcl-2) and diminished expression of Bcl-2 associated X protein (BAX) via PI3K/Akt signaling. BBR inhibited the proliferation of AA-FLS via promoting apoptosis through increased expression of BAX and diminished Bcl-2 transcription factor levels. Furthermore, T cells stimulated with IL-21 induced CD4+ CD196+ Th17 cells proliferation through RORγt activation mediated in a PI3K/Akt dependent manner. BBR inhibited the proliferation of Th17 cells through downregulation of RORγt in a concentration-dependent manner. BBR also promoted the differentiation of CD4+ CD25+ Treg cells through induction of forkhead box P3 (Foxp3) activation via aryl hydrocarbon receptor (AhR) and upregulation of cytochrome P450 family 1, subfamily A, polypeptide 1 (CYP1A1). Collectively, we conclude that BBR might attenuate AA-FLS proliferation through inhibition of IL-21/IL-21R dependent autophagy and regulates the Th17/Treg imbalance in RA.
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Antirreumáticos/farmacología , Artritis Reumatoide/patología , Autofagia/efectos de los fármacos , Berberina/farmacología , Sinoviocitos/efectos de los fármacos , Linfocitos T Reguladores/patología , Células Th17/patología , Animales , Proteínas Reguladoras de la Apoptosis/genética , Proteínas Reguladoras de la Apoptosis/metabolismo , Artritis Reumatoide/inducido químicamente , Artritis Reumatoide/inmunología , Diferenciación Celular/efectos de los fármacos , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Modelos Animales de Enfermedad , Silenciador del Gen , Interleucinas/metabolismo , Interleucinas/farmacología , Ratas , Ratas Wistar , Receptores de Hidrocarburo de Aril/genética , Receptores de Hidrocarburo de Aril/metabolismo , Receptores de Interleucina-21/metabolismo , Bazo/efectos de los fármacos , Bazo/inmunología , Sinoviocitos/patologíaRESUMEN
Inborn errors in interleukin 2 receptor, gamma (IL2RG) perturb signaling of the common gamma chain family cytokines and cause severe combined immunodeficiency (SCID). Here, we report two brothers suffering from chronic cryptosporidiosis, severe diarrhea, and cholangitis. Pan T, B, and NK cell numbers were normal, but immunophenotyping revealed defective B cell differentiation. Using whole exome sequencing, we identified a base pair deletion in the first exon of IL2RG predicted to cause a frameshift and premature stop. However, flow cytometry revealed normal surface expression of the IL-2Rγ chain. While IL-2, IL-7, and IL-15 signaling showed only mild defects of STAT5 phosphorylation in response to the respective cytokines, IL-4- and IL-21-induced phosphorylation of STAT3 and STAT6 was markedly reduced. Examination of RNA isoforms detected alternative splicing downstream of IL2RG exon 1 in both patients resulting in resolution of the predicted frameshift and 16 mutated amino acids. In silico modeling suggested that the IL-2Rγ mutation reduces the stabilization of IL-4 and IL-21 cytokine binding by affecting the N-terminal domain of the IL-2Rγ. Thus, our study shows that IL2RG deficiency can be associated with differential signaling defects. Confounding effects of alternative splicing may partially rescue genetic defects and should be considered in patients with inborn errors of immunity.
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Subunidad alfa del Receptor de Interleucina-21/genética , Inmunodeficiencia Combinada Grave/genética , Empalme Alternativo , Linfocitos B/inmunología , Preescolar , Colangitis/genética , Colangitis/inmunología , Croacia , Criptosporidiosis/genética , Criptosporidiosis/inmunología , Diarrea/genética , Diarrea/inmunología , Humanos , Subunidad alfa del Receptor de Interleucina-21/deficiencia , Subunidad alfa del Receptor de Interleucina-21/inmunología , Masculino , Infecciones del Sistema Respiratorio/genética , Infecciones del Sistema Respiratorio/inmunología , Inmunodeficiencia Combinada Grave/inmunologíaRESUMEN
BACKGROUND: We investigated the relationship between hepatitis B virus (HBV)-related pathogenesis and single nucleotide polymorphisms (SNPs) in interleukin-21 (IL-21)-JAK-STAT signaling pathway genes. METHODS: We used the high-resolution melting (HRM) method to genotype five SNPs (IL-21 rs2221903, IL-21 rs4833837, IL-21 receptor (IL-21R) rs2285452, JAK3 rs3008, and STAT3 rs1053023) in 546 HBV-infected patients and 353 healthy Chinese subjects. The HBV-infected patients were further divided into subgroups based on the HBV-related pathologies: chronic hepatitis B (CHB), HBV-related liver cirrhosis (LC), and HBV-related hepatocellular carcinoma (HCC). RESULTS: There were no significant differences in the genotype and allele distributions of the five SNPs between the HBV-infected patients and healthy subjects. The genotype and allele frequencies were similar in the two groups for IL-21 rs2221903 (A>G, P = 0.83 and 0.67), rs4833837 (A>G, P = 0.80 and 0.49), IL-21R rs2285452 (G>A, P = 0.25 and 0.68), STAT3 rs1053023 (A>G, P = 1.00 and 0.96), and JAK3 rs3008 (C>T, P = 0.32 and 0.54). However, patients with the IL-21R rs2285452 AA genotype were more susceptible to HBV-related HCC than those with the IL-21R rs2285452 GA/GG genotype (P = 0.03, OR = 3.27, 95% CI = 1.16-9.20). The serological marker model of "HBsAg+, HBeAg+, HBcAb+" was predominant among patients with HBV infection. However, there was no association between the genotype's distribution of the five SNPs and the serological marker models (P > 0.05). CONCLUSIONS: These findings demonstrate that the IL-21R rs2285452 AA genotype increases the risk of HBV-related HCC in Chinese patients.
Asunto(s)
Carcinoma Hepatocelular/genética , Hepatitis B Crónica/genética , Subunidad alfa del Receptor de Interleucina-21/genética , Neoplasias Hepáticas/genética , Polimorfismo de Nucleótido Simple , Adulto , Pueblo Asiatico/genética , Carcinoma Hepatocelular/virología , Estudios de Casos y Controles , Femenino , Frecuencia de los Genes , Antígenos de la Hepatitis B/sangre , Antígenos de la Hepatitis B/genética , Hepatitis B Crónica/complicaciones , Humanos , Cirrosis Hepática/genética , Cirrosis Hepática/virología , Neoplasias Hepáticas/virología , Masculino , Persona de Mediana EdadRESUMEN
Human immunodeficiency virus (HIV) infection is characterized by dysfunctional B cell responses. Here we show that chronic simian immunodeficiency virus (SIV) infection is characterized by an expansion of either lymph node germinal center (GC) B cells that co-express Bcl6, Ki-67 and IL-21R and correlate with expanded T follicular helper (Tfh) cells or B cells that lack Bcl6, Ki-67 and IL-21R but express high levels of anti-apoptotic Bcl2 that negatively correlate with Tfh cells. The lack of Tfh cells likely contributes to persistence of dysfunctional non-proliferating B cells during chronic infection. These findings have implications for protective immunity in HIV-infected individuals who harbour low frequencies of Tfh cells.
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Linfocitos B/inmunología , Infecciones por VIH/inmunología , Síndrome de Inmunodeficiencia Adquirida del Simio/inmunología , Virus de la Inmunodeficiencia de los Simios/genética , Animales , Linfocitos B/patología , Diferenciación Celular/inmunología , Centro Germinal/inmunología , Centro Germinal/virología , Infecciones por VIH/genética , Infecciones por VIH/patología , Infecciones por VIH/virología , Humanos , Ganglios Linfáticos/inmunología , Ganglios Linfáticos/patología , Macaca mulatta/inmunología , Fenotipo , Proteínas Proto-Oncogénicas c-bcl-2/genética , Proteínas Proto-Oncogénicas c-bcl-6/genética , Síndrome de Inmunodeficiencia Adquirida del Simio/genética , Síndrome de Inmunodeficiencia Adquirida del Simio/patología , Síndrome de Inmunodeficiencia Adquirida del Simio/virología , Virus de la Inmunodeficiencia de los Simios/inmunología , Virus de la Inmunodeficiencia de los Simios/patogenicidad , Linfocitos T Colaboradores-Inductores/inmunología , Linfocitos T Colaboradores-Inductores/patologíaRESUMEN
Interleukin 21 (IL-21) is a pleiotropic common cytokine receptor γ chain cytokine that promotes the effector functions of NK cells and CD8+ T cells and inhibits CD8+ T cell exhaustion during chronic infection. We found that the absolute number of short-lived effector CD8+ T cells (SLECs) (KLRG1high CD127low) decreased significantly in IL-21 receptor-deficient (IL-21R-/-) mice during Mycobacterium bovis bacillus Calmette-Guérin (BCG) infection. Early effector CD8+ T cells (EECs) (KLRG1low CD127low) were normally generated in IL-21R-/- mice after infection. Exhausted CD8+ T cells (PD-1high KLRG1low) were also normally generated in IL-21R-/- mice after infection. Mixed bone marrow (BM) chimera and transfer experiments showed that IL-21R on CD8+ T cells was essential for the proliferation of EECs, allowing them to differentiate into SLECs after BCG infection. On the other hand, the number of SLECs increased significantly after infection with recombinant BCG (rBCG) that secreted an antigen 85B (Ag85B)-IL-21 fusion protein (rBCG-Ag85B-IL-21), but the number of exhausted CD8+ T cells did not change after rBCG-Ag85B-IL-21 infection. These results suggest that IL-21 signaling drives the differentiation of SLECs from EECs but does not inhibit the exhaustion of CD8+ T cells following BCG infection in mice.
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Linfocitos T CD8-positivos/inmunología , Interleucinas/metabolismo , Mycobacterium bovis/inmunología , Subgrupos de Linfocitos T/inmunología , Tuberculosis/inmunología , Animales , Linfocitos T CD8-positivos/química , Diferenciación Celular , Modelos Animales de Enfermedad , Inmunofenotipificación , Subunidad alfa del Receptor de Interleucina-7/análisis , Lectinas Tipo C , Ratones Endogámicos C57BL , Ratones Noqueados , Receptores Inmunológicos/análisis , Receptores de Interleucina-21/análisis , Receptores de Interleucina-21/deficiencia , Subgrupos de Linfocitos T/químicaRESUMEN
Rheumatoid arthritis (RA) is a systemic autoimmune inflammatory disorder designated with hyperplastic synovium, bone destruction and cartilage degradation. Current therapies involve targeting major cytokines and inflammatory mediators involved in RA to alleviate the pain and provide a temporary relief. Interleukin 21 (IL-21), a recently identified cytokine is known to possess a versatile role in modulating the cells of the RA synovium. Over the past decade, the pleiotropic role of IL-21 in RA pathogenesis has been implicated in several aspects. T helper 17 (Th17) and follicular T helper cells (Tfh), being the key immunomodulators of the RA synovium secrete high amounts of IL-21 during disease progression. Several studies have provided experimental evidences elucidating the multifaceted role of IL-21 in RA disease progression. IL-21 has the potential to activate T cells, B cells, monocytes/macrophages and synovial fibroblasts in RA pathogenesis through activation of JAK-STAT, MAPK and PI3K/Akt signaling pathways. Till date, therapies targeting Th17 cells and its inflammatory cytokines have been under investigation and are subjected to various clinical trials. This review showcases the role of IL-21 in RA pathogenesis and recent reports implicating its function in various immune cells, major signaling pathways, and in promoting osteoclastogenesis.
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Artritis Reumatoide/tratamiento farmacológico , Artritis Reumatoide/metabolismo , Fibroblastos/metabolismo , Interleucinas/metabolismo , Animales , Cartílago/metabolismo , Citocinas/metabolismo , Humanos , Interleucinas/inmunología , Fosfatidilinositol 3-Quinasas/metabolismoRESUMEN
The current study investigated the therapeutic effect of berberine (BBR), an alkaloid derivative against IL-21/IL-21R mediated phosphotidyl inositol 3 kinase/protein kinase B (PI3K/Akt) signaling in adjuvant induced arthritic fibroblast-like synoviocytes (AA-FLS) isolated from rats and IL-21 mediated osteoclastogenesis in bone-marrow derived monocytes (BMMs). BBR (15-45⯵M) treatment attenuated the gene and protein levels of IL-21R complex. BBR suppressed the levels of IL-21 (20â¯ng/ml) mediated production of inflammatory cytokines such as: tumor necrosis factor alpha (TNFα), interleukin 1 beta (IL-1ß), interleukin 6 (IL-6) and interleukin 23 (IL-23) in AA-FLS cells. Subsequently, BBR ameliorated the gene and protein expression levels of mechanistic target of rapamycin (mTOR), IL-23 and nuclear factor kappa B (NFκB) p65 through the inhibition of PI3K and upregulation of phosphatase and tensin homolog (PTEN) at the protein level. Furthermore, BBR also inhibited the phosphorylation of Akt and NFκB p65 in a dose dependant manner. LY294002 (20⯵M) treatment suppressed the PI3K/Akt signaling and its downstream elements in AA-FLS cells. BBR also modulated IL-21 mediated osteoclastogenesis through the suppression of PI3K dependant nuclear factor of activated T-cells 1 (Nfatc1) induction. Moreover, BBR controlled the osteoclast differentiation via inhibition of various bone resorptive enzymes including: cathepsin K, matrix metalloproteinase 9 (MMP9) and tartarate acid phosphatase (TRAP). LY294002 also inhibited osteoclast formation via suppression of PI3K mediated Nfatc1 induction and other downstream elements. Overall, our findings suggest that BBR is a potential candidate for therapeutic targeting of IL-21/IL-21R mediated RA pathogenesis.
Asunto(s)
Berberina/farmacología , Fibroblastos/patología , Inflamación/patología , Interleucinas/metabolismo , Osteogénesis/efectos de los fármacos , Receptores de Interleucina-21/metabolismo , Transducción de Señal/efectos de los fármacos , Sinoviocitos/patología , Animales , Artritis Experimental/genética , Artritis Experimental/patología , Berberina/química , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Citocinas/metabolismo , Femenino , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Mediadores de Inflamación/metabolismo , Masculino , Metaloproteinasa 9 de la Matriz/metabolismo , Modelos Biológicos , Factores de Transcripción NFATC/metabolismo , Osteogénesis/genética , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas Wistar , Receptores de Interleucina-21/genética , Sinoviocitos/efectos de los fármacos , Sinoviocitos/metabolismoRESUMEN
OBJECTIVES: Granulomatosis with polyangiitis (GPA) is a relapsing small-vessel vasculitis characterized by circulating ANCA against PR3. The mechanisms that trigger PR3-ANCA production are unknown. The aim of this study was to determine whether endogenous factors [B cell activating factor (BAFF) and IL-21] and exogenous factors [oligodeoxynucleotides containing CpG motifs (CpG-ODN)] synergize in stimulating PR3-ANCA production in GPA patients. METHODS: Peripheral blood mononuclear cells from GPA patients and healthy controls (HCs) were cultured in the presence of BAFF and IL-21, with or without CpG-ODN, for 12 days. PR3-ANCA production in culture supernatants was quantified by Phadia EliA. Phenotypic characterization and the influence of CpG-ODN treatment on IL-21 receptor (IL-21R), transmembrane activator and calcium-modulator and cyclophilin ligand interactor (TACI) and BAFF receptor (BAFF-R) expression on B cells was analysed by flow cytometry. RESULTS: Stimulation with BAFF and IL-21 significantly increased ANCA production in patient samples, which could be augmented further by addition of CpG-ODN. Stimulation with CpG-ODN increased the percentage of IL-21R(+) and TACI(+) B cells but did not affect BAFF-R expression. GPA patients had an increased percentage of circulating IL-21R(+) and a decreased percentage of TACI(+) circulating memory B cells when compared with HCs. Additionally, patients had decreased expression of BAFF-R on B cells, which was inversely correlated with BAFF concentrations in plasma. CONCLUSION: Our data demonstrate that endogenous and exogenous factors can synergize to promote PR3-ANCA production. Mechanistically, CpG-ODN up-regulated IL-21R and TACI expression on B cells, possibly sensitizing these cells for IL-21- and BAFF-mediated signals. Agents inhibiting Toll-like receptor 9, BAFF and IL-21 signalling pathways may serve as potential therapeutics for intervention in GPA patients.
Asunto(s)
Autoanticuerpos/inmunología , Factor Activador de Células B/metabolismo , Granulomatosis con Poliangitis/inmunología , Interleucinas/metabolismo , Activación de Linfocitos/inmunología , Mieloblastina/inmunología , Receptor Toll-Like 9/metabolismo , Adulto , Anciano , Linfocitos B/inmunología , Linfocitos B/metabolismo , Linfocitos B/patología , Proliferación Celular , Ensayo de Inmunoadsorción Enzimática , Femenino , Citometría de Flujo , Granulomatosis con Poliangitis/metabolismo , Granulomatosis con Poliangitis/patología , Humanos , Masculino , Persona de Mediana Edad , Mieloblastina/metabolismo , Receptor Toll-Like 9/inmunologíaRESUMEN
T helper cells 17 (Th17) are recognized as key participants in the pathogenesis of chronic autoimmune diseases such as multiple sclerosis (MS). Regulation of Th17 differentiation is a valuable strategy for diagnosis and treatment of these complicated immune disorders. Here, by genome-wide expression profiling of microRNAs (miRs), we screened miR-30a, whose level was greatly decreased during Th17 differentiation and the process of demyelination disease, both in MS patients and experimental autoimmune encephalomyelitis (EAE) mice. Enforced constitutive expression of miR-30a in naïve T cells inhibited their differentiation into Th17, and in vivo overexpression of miR-30a resulted in fewer Th17 and alleviative EAE. Moreover, target prediction analysis and dual luciferase report assay revealed that interleukin-21 receptor (IL-21R) was a direct target of miR-30a, a finding consistent with the results that miR-30a downregulated the expression of IL-21R, while overexpression of IL-21R alleviated the inhibitory effect of miR-30a on Th17 differentiation. Taken together, our findings imply that miR-30a inhibits Th17 differentiation and the pathogenesis of MS by targeting IL-21R.
Asunto(s)
Diferenciación Celular , Encefalomielitis Autoinmune Experimental/metabolismo , Subunidad alfa del Receptor de Interleucina-21/metabolismo , MicroARNs/metabolismo , Esclerosis Múltiple/metabolismo , Células Th17 , Adulto , Animales , Humanos , Ratones , Ratones Endogámicos C57BLRESUMEN
To determine the potential role of interleukin-21 (IL-21) / IL-21 receptor (IL-21R) in the pathogenesis of autoimmune thyroid disease (AITD) mainly known as Graves' disease (GD) and Hashimoto's thyroiditis (HT). IL-21 and IL-21R of peripheral blood samples and/or thyroid tissues from AITD patients and healthy controls were analyzed by ELISA, quantitative real-time polymerase chain reaction (qRT-PCR), flow cytometry and immunohistochemistry. In vitro, the mRNA and protein of inflammatory cytokines of cultured peripheral blood mononuclear cells (PBMCs) upon recombinant human IL-21 (rhIL-21) stimulation were detected. There was an increased serum concentration of IL-21 in untreated GD and HT patients, and IL-21(+)CD3(+)CD8(-)T cells were significantly increased in PBMCs of HT patients compared with healthy volunteers. The IL-21 mRNA expression in PBMCs increased dramatically in GD and HT patients, and marked augmentations of IL-21 and IL-21R mRNA in thyroid tissues of HT patients were observed. Immunohistochemical staining confirmed the expression of IL-21R protein in HT thyroid cells and lymphocytes. In vitro, PBMCs from GD cultured with rhIL-21 induced increased IL-17A but decreased IL-4 production, while from HT stimulated by rhIL-21 induced augmented production of IFN-γ. In conclusion, the expression of IL-21 and IL-21R were up-regulated in AITD and may be involved in the pathogenesis of the disease through augmenting aberrant immune cascade.
Asunto(s)
Enfermedad de Graves/inmunología , Enfermedad de Hashimoto/inmunología , Interleucinas/metabolismo , Receptores de Interleucina-21/metabolismo , Adulto , Citometría de Flujo , Humanos , Inmunohistoquímica , Interferón gamma , Interleucina-17/metabolismo , Interleucina-4/metabolismo , Interleucinas/genética , Leucocitos Mononucleares/efectos de los fármacos , Leucocitos Mononucleares/inmunología , Masculino , Persona de Mediana Edad , Reacción en Cadena en Tiempo Real de la Polimerasa , Receptores de Interleucina-21/genética , Proteínas Recombinantes/administración & dosificación , Glándula Tiroides/inmunología , Glándula Tiroides/ultraestructuraRESUMEN
BACKGROUND: Interleukin-21 (IL-21) has been implicated in the development of Th2-mediated immune responses; however, the exact role it plays in allergic diseases is not well understood. OBJECTIVE: To elucidate the contribution of IL-21 receptor signalling to Th2-dependent immune responses in the lung. METHODS: We compared allergic airway responses in wild-type BALB/c and Il21r-deficient mice exposed to local airway challenge with house dust mite (HDM). RESULTS: We demonstrate that IL-21R-deficiency reduces HDM-driven airway hyperresponsiveness (AHR) with only partial effects on airway inflammation. Concomitant with the reduction in AHR in Il21r-deficient mice, significant suppression was observed in protein levels of the Th2 cytokines IL-4, and IL-13. In contrast, IL-21R-deficiency was associated with an increase in PBS- and allergen-driven IgE levels, while IgG1 and IgG2a levels were decreased. Moreover, our results suggest that IL-21 may contribute to AHR through its ability to both directly induce Th2 cell survival and to impair regulatory T-cell suppression of Th2 cytokine production. Importantly, we show that IL-21-positive cells are increased in the bronchial mucosa of asthmatics compared with non-asthmatics. CONCLUSION: These results suggest that IL-21 plays an important role in the allergic diathesis by enhancing Th2 cytokine production through multiple mechanisms including the suppression of Treg inhibitory effects on Th2 cell cytokine production.
Asunto(s)
Hipersensibilidad/inmunología , Hipersensibilidad/metabolismo , Receptores de Interleucina-21/metabolismo , Transducción de Señal , Células Th2/inmunología , Células Th2/metabolismo , Alérgenos/inmunología , Animales , Hiperreactividad Bronquial/genética , Hiperreactividad Bronquial/inmunología , Hiperreactividad Bronquial/metabolismo , Citocinas/metabolismo , Modelos Animales de Enfermedad , Femenino , Hipersensibilidad/genética , Inmunoglobulina E/inmunología , Inmunoglobulina E/metabolismo , Masculino , Ratones , Ratones Noqueados , Receptores de Interleucina-21/deficiencia , Receptores de Interleucina-21/genética , Linfocitos T Reguladores/inmunología , Linfocitos T Reguladores/metabolismoRESUMEN
BACKGROUND: The capacity of CD8(+) T cells to control infections and mediate antitumor immunity requires the development and survival of effector and memory cells. IL-21 has emerged as a potent inducer of CD8(+) T-cell effector function and memory development in mouse models of infectious disease. However, the role of IL-21 and associated signaling pathways in protective CD8(+) T-cell immunity in human subjects is unknown. OBJECTIVE: We sought to determine which signaling pathways mediate the effects of IL-21 on human CD8(+) T cells and whether defects in these pathways contribute to disease pathogenesis in patients with primary immunodeficiencies caused by mutations in components of the IL-21 signaling cascade. METHODS: Human primary immunodeficiencies resulting from monogenic mutations provide a unique opportunity to assess the requirement for particular molecules in regulating human lymphocyte function. Lymphocytes from patients with loss-of-function mutations in signal transducer and activator of transcription 1 (STAT1), STAT3, or IL-21 receptor (IL21R) were used to assess the respective roles of these genes in human CD8(+) T-cell differentiation in vivo and in vitro. RESULTS: Mutations in STAT3 and IL21R, but not STAT1, led to a decrease in multiple memory CD8(+) T-cell subsets in vivo, indicating that STAT3 signaling, possibly downstream of IL-21R, regulates the memory cell pool. Furthermore, STAT3 was important for inducing the lytic machinery in IL-21-stimulated naive CD8(+) T cells. However, this defect was overcome by T-cell receptor engagement. CONCLUSION: The IL-21R/STAT3 pathway is required for many aspects of human CD8(+) T-cell behavior but in some cases can be compensated by other signals. This helps explain the relatively mild susceptibility to viral disease observed in STAT3- and IL-21R-deficient subjects.
Asunto(s)
Linfocitos T CD8-positivos/citología , Diferenciación Celular , Memoria Inmunológica , Síndrome de Job/genética , Mutación , Factor de Transcripción STAT3/metabolismo , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/metabolismo , Diferenciación Celular/genética , Diferenciación Celular/inmunología , Humanos , Interleucinas/genética , Interleucinas/inmunología , Interleucinas/metabolismo , Síndrome de Job/inmunología , Síndrome de Job/patología , Receptores de Interleucina-21/genética , Receptores de Interleucina-21/inmunología , Receptores de Interleucina-21/metabolismo , Factor de Transcripción STAT3/genéticaRESUMEN
AIMS: Aimed to identify a new susceptibility gene associated with primary biliary cholangitis (PBC) in Chinese Han and investigate the possible mechanism of that gene in PBC. METHODS: A total of 466 PBC and 694 healthy controls (HC) were included in our study, and genotyping GTF2I gene variants by Sequenom. CD19 + B cells were isolated for Chromatin immunoprecipitation sequencing (ChIP-seq). Additionally, MEME-ChIP was utilized to perform searches for known motifs and de novo motif discovery. The GTF2I ChIP-seq of hematopoietic cell line (K562) results were obtained from ENCODE (GSE176987, GSE177691). The Genomic HyperBrowser was used to determine overlap and hierarchal clustering between ours and ENCODE datasets. RESULTS: The frequency of the rs117026326 variant T allele was significantly higher in PBC patients than that in HC (20.26% compared with 13.89%, Pc = 1.09E-04). Furthermore, we observed an elevated proportion of GTF2I binding site located in the upstream and 5' UTR of genes in PBC in comparison with HC. Additionally, an in-depth analysis of IL21R region revealed that GTF2I might bind to the IL21R promoter to regulate the expression of the IL21R, with four peaks of GTF2I binding sites, including three increased binding sites in upstream, one increased binding site in 5' UTR. Motif analysis by MEME-ChIP uncovered five significant motifs. A significant overlap between our ChIP and GSE176987, GSE17769 were found by the Genomic HyperBroswer. CONCLUSIONS: Our study confirmed that GTF2I was associated with PBC in Chinese Han. Furthermore, our gene function analysis indicated that IL21R may be the target gene regulated by GTF2I.
Asunto(s)
Cirrosis Hepática Biliar , Factores de Transcripción TFIII , Factores de Transcripción TFII , Humanos , Regiones no Traducidas 5' , China , Secuenciación de Inmunoprecipitación de Cromatina , Cirrosis Hepática Biliar/genética , Receptores de Interleucina-21/genética , Factores de Transcripción TFII/genética , Factores de Transcripción TFIII/genéticaRESUMEN
Some benign and malignant breast tumours are similar in pathological morphology, which are difficult to be distinguished in clinical diagnosis. In this study, we intended to explore novel biomarkers for differential diagnosis of benign and malignant breast tumours. Methylation EPIC 850K beadchip and RNA-sequencing were used to analyse 29 tissue samples from patients with early-stage breast cancer (BC) and benign breast tumours for differently methylated and expressed genes. The altered methylation of IL21R was semi-quantitatively validated in an independent study with 566 tissue samples (279 BC vs. 287 benign breast tumours) using mass spectrometry. Binary logistic regression analysis was performed to evaluate the association between IL21R methylation and BC. BC-associated IL21R hypomethylation and overexpression were identified in the discovery round. In the validation round, BC patients presented significant IL21R hypomethylation compared to women with benign breast tumours (ORs ≥1.29 per-10% methylation, p-values ≤ 5.69E-14), and this hypomethylation was even enhanced in BC patients with ER-negative and PR-negative tumours as well as with triple-negative tumours. The methylation of IL21R showed efficient discriminatory power to distinguish benign breast tumours from BC (area under curve (AUC) = 0.88), and especially from ER-negative BC (AUC = 0.95), PR-negative BC (AUC = 0.93) and triple-negative BC (AUC = 0.96). We disclosed significant IL21R hypomethylation in patients with BC compared to women with benign breast tumours, and revealed the somatic change of DNA methylation could be a potential biomarker for molecular pathology of BC.
Asunto(s)
Biomarcadores de Tumor , Neoplasias de la Mama , Metilación de ADN , Femenino , Humanos , Biomarcadores de Tumor/genética , Biomarcadores de Tumor/metabolismo , Neoplasias de la Mama/genética , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Neoplasias de la Mama/diagnóstico , Diagnóstico Diferencial , Subunidad alfa del Receptor de Interleucina-21 , Receptores de Interleucina/genética , Receptores de Interleucina/metabolismoRESUMEN
Rheumatoid Arthritis (RA) is characterized by joint destruction, chronic inflammation, and autoantibody production. IL-21/IL-21R plays an essential role in the immunopathology of RA. Elevated IL-21 serum levels have been associated with RA and disease activity. Here, we evaluated the association of IL-21/IL-21R polymorphisms and IL-21 serum levels with RA. The study included 275 RA patients and 280 Control subjects (CSs). Single nucleotide polymorphisms IL-21 (rs2055979 and rs2221903) and IL-21R (rs3093301) were genotyped using PCR-RFLP. Clinical activity was evaluated by DAS28-ESR; IL-21 and anti-CCP serum levels were quantified by ELISA. The IL-21 rs2055979 AA genotype was higher in RA patients than in the CS group (p = 0.0216, OR = 1.761, 95% CI = 1.085-2.859); furthermore, RA patients showed anti-CCP elevated levels compared to the CA genotype (p = 0.0296). The IL21R rs3093301 AA genotype was also higher in RA patients than in the CS group (p = 0.0122, OR = 1.965, 95% CI = 1.153-3.348). The AT haplotypes of IL-21 rs2055979 and rs2221903 were more frequent (49%) in the RA group (p = 0.006). IL-21 serum levels were significantly elevated in the RA group, but without an association with IL-21 polymorphisms. In conclusion, IL-21 rs2255979 and IL-21R rs3093301 are associated with a higher risk of RA, and could be a genetic marker. Moreover, the elevated IL-21 levels in RA suggest that IL-21/IL-21R could be a therapeutic target in RA.