RESUMO
Bullous pemphigoid (BP) is a type 2 inflammation- and immunity-driven skin disease, yet a comprehensive understanding of the immune landscape, particularly immune-stromal crosstalk in BP, remains elusive. Herein, using single-cell RNA sequencing (scRNA-seq) and in vitro functional analyzes, we pinpoint Th2 cells, dendritic cells (DCs), and fibroblasts as crucial cell populations. The IL13-IL13RA1 ligand-receptor pair is identified as the most significant mediator of immune-stromal crosstalk in BP. Notably, fibroblasts and DCs expressing IL13RA1 respond to IL13-secreting Th2 cells, thereby amplifying Th2 cell-mediated cascade responses, which occurs through the specific upregulation of PLA2G2A in fibroblasts and CCL17 in myeloid cells, creating a positive feedback loop integral to immune-stromal crosstalk. Furthermore, PLA2G2A and CCL17 contribute to an increased titer of pathogenic anti-BP180-NC16A autoantibodies in BP patients. Our work provides a comprehensive insight into BP pathogenesis and shows a mechanism governing immune-stromal interactions, providing potential avenues for future therapeutic research.
Assuntos
Quimiocina CCL17 , Células Dendríticas , Fibroblastos , Penfigoide Bolhoso , Análise de Célula Única , Células Th2 , Humanos , Penfigoide Bolhoso/imunologia , Penfigoide Bolhoso/genética , Análise de Célula Única/métodos , Fibroblastos/metabolismo , Fibroblastos/imunologia , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Quimiocina CCL17/genética , Quimiocina CCL17/metabolismo , Células Th2/imunologia , Autoanticorpos/imunologia , Transcriptoma , Interleucina-13/metabolismo , Interleucina-13/genética , Interleucina-13/imunologia , Colágenos não Fibrilares/imunologia , Colágenos não Fibrilares/genética , Colágenos não Fibrilares/metabolismo , Inflamação/imunologia , Inflamação/genética , Inflamação/metabolismo , Perfilação da Expressão Gênica/métodos , Masculino , Feminino , Autoantígenos/imunologia , Autoantígenos/metabolismo , Autoantígenos/genética , Colágeno Tipo XVII , Células Mieloides/metabolismo , Células Mieloides/imunologia , Células Estromais/metabolismo , Células Estromais/imunologiaRESUMO
OBJECTIVES: Alveolar echinococcosis (AE) represents one of the deadliest helminthic infections, characterized by an insidious onset and high lethality. METHODS: This study utilized the Gene Expression Omnibus (GEO) database, applied Weighted Correlation Network Analysis (WGCNA) and Differential Expression Analysis (DEA), and employed the Matthews Correlation Coefficient (MCC) to identify CCL17 and CCL19 as key genes in AE. Immunohistochemistry and immunofluorescence co-localization techniques were used to examine the expression of CCL17 and CCL19 in liver tissue lesions of AE patients. Additionally, a mouse model of multilocular echinococcus larvae infection was developed to study the temporal expression patterns of these genes, along with liver fibrosis and inflammatory responses. RESULTS: The in vitro model simulating echinococcal larva infection mirrored the hepatic microenvironment post-infection with multilocular echinococcal tapeworms. Quantitative RT-PCR analysis showed that liver fibrosis occurred in AE patients, with proximal activation and increased expression of CCL17 and CCL19 over time post-infection. Notably, expression peaked during the late stages of infection. Similarly, F4/80, a macrophage marker, exhibited corresponding trends in expression. Upon stimulation of normal hepatocytes by vesicular larvae in cellular experiments, there was a significant increase in CCL17 and CCL19 expression at 12 h post-infection, mirroring the upregulation observed with F4/80. CONCLUSION: CCL17 and CCL19 facilitate macrophage aggregation via the chemokine pathway and their increased expression correlates with the progression of infection, suggesting their potential as biomarkers for AE progression.
Assuntos
Biomarcadores , Quimiocina CCL17 , Quimiocina CCL19 , Progressão da Doença , Animais , Humanos , Camundongos , Biomarcadores/metabolismo , Quimiocina CCL19/metabolismo , Quimiocina CCL17/metabolismo , Quimiocina CCL17/genética , Equinococose/metabolismo , Cirrose Hepática/parasitologia , Cirrose Hepática/metabolismo , Cirrose Hepática/patologia , Modelos Animais de Doenças , Fígado/parasitologia , Fígado/metabolismo , Fígado/patologia , Equinococose Hepática/metabolismo , Equinococose Hepática/parasitologia , Feminino , Masculino , Hepatócitos/metabolismo , Hepatócitos/parasitologiaRESUMO
BACKGROUND: Atopic dermatitis (AD) is a common and recurrent inflammatory disease with strong genetic susceptibility. The abnormal production of chemokines plays an important role in the occurrence and development of AD. METHODS: A comprehensive online literature search was performed in databases of China National Knowledge Infrastructure, Wanfang, VIP China Science and Technology Journal Database, China Biomedical Literature Database, PubMed, Embase and Cochrane Library to retrieve relevant articles published from January 2000 to October 2022. The odds ratio (OR) with its 95% confidence interval (CI) was employed to calculate this relationship. RESULTS: A total of 7 studies were finally screened out, including 1316 AD patients and 1099 controls. There were 3 studies for CC chemokine ligand 5 (CCL5) polymorphisms, 2 for CCL11 polymorphisms, and 2 for CCL17 polymorphisms, respectively. The meta-analysis revealed a significant association between the CCL5â -â 403G/A polymorphism and AD under the allelic model (A vs G: ORâ =â 1.25, 95% CIâ =â 1.02-1.52, Pâ =â .03), heterozygous model (AG vs GG: ORâ =â 1.40, 95% CIâ =â 1.08-1.80, Pâ =â .01) and dominant model (AAâ +â AG vs GG: ORâ =â 1.38, 95% CIâ =â 1.08-1.76, Pâ =â .01) in a fixed-effect model. The allelic model (G vs C: ORâ =â 1.46, 95% CIâ =â 1.07-1.98, Pâ <â .01) and dominant model (GGâ +â GC vs CC: ORâ =â 1.74, 95% CIâ =â 1.23-2.47, Pâ <â .001) of the CCL5â -â 28C/G polymorphism were also associated with an increased risk of AD. However, this significant association was not found in other alleles and genotypes (Pâ >â .05). CONCLUSION: Our results show that the A allele, AG and AAâ +â AG genotypes of the CCL5â -â 403G/A polymorphism, the G allele and GGâ +â GC genotype of the CCL5â -â 28C/G polymorphism are risk factors for AD. Future studies with large population are still needed to further explore those correlations.
Assuntos
Quimiocina CCL11 , Quimiocina CCL17 , Quimiocina CCL5 , Dermatite Atópica , Humanos , Quimiocina CCL11/genética , Quimiocina CCL17/genética , Quimiocina CCL5/genética , Dermatite Atópica/genética , Predisposição Genética para Doença , Genótipo , Ligantes , Polimorfismo de Nucleotídeo Único , Fatores de RiscoRESUMO
Background/Aims: Chemokines are known to play critical roles mediating inflammation in many pathophysiological processes. The aim of this study was to investigate the role of chemokine receptor CCR4 and its ligands CCL17 and CCL22 in human morbid obesity. Methods: Circulating levels of CCL17 and CCL22 were measured in 60 morbidly obese patients (mean age, 45 ± 1 years; body mass index/BMI, 44 ± 1 kg/m2) who had undergone bariatric bypass surgery, and 20 control subjects. Paired subcutaneous (SCAT) and visceral adipose tissue (VCAT) from patients were analysed to measure expression of CCR4 and its ligands by RT-PCR, western blot and immunohistochemical analysis. The effects of CCR4 neutralization ex vivo on leukocyte-endothelial cells were also evaluated. Results: Compared with controls, morbidly obese patients presented higher circulating levels of CCL17 (p=0.029) and CCL22 (p<0.001) and this increase was positively correlated with BMI (p=0.013 and p=0.0016), and HOMA-IR Index (p=0.042 and p< 0.001). Upregulation of CCR4, CCL17 and CCL22 expression was detected in VCAT in comparison with SCAT (p<0.05). Using the parallel-plate flow chamber model, blockade of endothelial CCR4 function with the neutralizing antibody anti-CCR4 in morbidly obese patients significantly reduced leucocyte adhesiveness to dysfunctional endothelium, a key event in atherogenesis. Additionally, CCL17 and CCL22 increased activation of the ERK1/2 mitogen-activated protein kinase signalling pathway in human aortic endothelial cells, which was significantly reduced by CCR4 inhibition (p=0.016 and p<0.05). Conclusion: Based on these findings, pharmacological modulation of the CCR4 axis could represent a new therapeutic approach to prevent adipose tissue dysfunction in obesity.
Assuntos
Células Endoteliais , Obesidade Mórbida , Humanos , Adulto , Pessoa de Meia-Idade , Células Endoteliais/metabolismo , Obesidade Mórbida/complicações , Obesidade Mórbida/cirurgia , Quimiocina CCL17/genética , Quimiocinas , Transdução de Sinais , Receptores de Quimiocinas/metabolismo , Quimiocina CCL22/genéticaRESUMO
BACKGROUND: Chemokines are critical players in the local immune responses to tumors. CCL17 (thymus and activation-regulated chemokine, TARC) and CCL22 (macrophage-derived chemokine, MDC) can attract CCR4-bearing cells involving the immune landscape of cancer. However, their direct roles and functional states in tumors remain largely unclear. METHODS: We analyzed the lymphoma-related scRNA-seq and bulk RNA-seq datasets and identified the CCL17/CCL22-CCR4 axis as the unique participant of the tumor microenvironment. Then we edited the A20 lymphoma cell line to express CCL17 and CCL22 and assessed their function using three mouse models (Balb/C mouse, Nude mouse, and NSG mouse). In addition, we retrospectively checked the relationship between the CCL17/CCL22-CCR4 axis and the survival rates of cancer patients. RESULTS: The active CCL17/CCL22-CCR4 axis is a distinctive feature of the Hodgkin lymphoma microenvironment. CCR4 is widely expressed in immune cells but highly exists on the surface of NK, NKT, and Treg cells. The tumor model of Balb/C mice showed that CCL17 acts as an anti-tumor chemokine mediated by activated T cell response. In addition, the tumor model of Nude mice showed that CCL17 recruits NK cells for inhibiting lymphoma growth and enhances the NK-cDC1 interaction for resisting IL4i1-mediated immunosuppression. Interestingly, CCL17-mediated antitumor immune responses depend on lymphoid lineages but not mainly myeloid ones. Furthermore, we found CCL17/CCL22-CCR4 axis cannot be regarded as biomarkers of poor prognosis in most cancer types from the TCGA database. CONCLUSION: We provided direct evidence of antitumor functions of CCL17 mediated by the recruitment of conventional T cells, NKT cells, and NK cells. Clinical survival outcomes of target gene (CCL17, CCL22, and CCR4) expression also identified that CCL17/CCL22-CCR4 axis is not a marker of poor prognosis.
Assuntos
Quimiocina CCL17 , Quimiocinas , Humanos , Camundongos , Animais , Quimiocina CCL17/genética , Quimiocina CCL17/metabolismo , Camundongos Nus , Estudos Retrospectivos , Linfócitos/metabolismo , Receptores CCR4/genética , Receptores CCR4/metabolismo , L-Aminoácido OxidaseRESUMO
BACKGROUND: T cell lymphoma is a complex and highly aggressive clinicopathological entity with a poor outcome. The angioimmunoblastic T-cell lymphoma (AITL) tumor immune microenvironment is poorly investigated. METHODS: Here, to the best of our knowledge, spatial transcriptomics was applied for the first time to study AITL. RESULTS: Using this method, we observed that AITL was surrounded by cells bearing immune-suppressive markers. CCL17 and CCL22, the dominant ligands for CCR4, were up-regulated, while the expression of natural killer (NK) cell and CD8+ cytotoxic T lymphocyte (CTL) markers decreased. Colocalization of Treg cells with the CD4+ TFH-GC region was also deduced from the bioinformatic analysis. The results obtained with spatial transcriptomics confirm that AITL has a suppressive immune environment. Chemotherapy based on the CHOP regimen (cyclophosphamide, doxorubicin, vincristine plus prednisone) induced complete remission (CR) in this AITL patient. However, the duration of remission (DoR) remains a concern. CONCLUSIONS: This study demonstrates that AITL has an immune suppressive environment and suggests that anti-CCR4 therapy could be a promising treatment for this lethal disease.
Assuntos
Linfadenopatia Imunoblástica , Linfoma de Células T , Quimiocina CCL17/genética , Quimiocina CCL17/uso terapêutico , Quimiocina CCL22/genética , Quimiocina CCL22/uso terapêutico , Ciclofosfamida/uso terapêutico , Doxorrubicina/uso terapêutico , Humanos , Linfadenopatia Imunoblástica/tratamento farmacológico , Linfadenopatia Imunoblástica/genética , Linfadenopatia Imunoblástica/patologia , Linfoma de Células T/tratamento farmacológico , Linfoma de Células T/genética , Linfoma de Células T/patologia , Prednisona/uso terapêutico , Transcriptoma , Microambiente Tumoral/genética , Vincristina/uso terapêuticoRESUMO
Abacavir (ABC)-induced hypersensitivity (AHS) is strongly associated with human leukocyte antigen (HLA)-B*57 : 01 expression. Previous studies have demonstrated the feasibility of applying the HLA-transgenic mouse model in this context. ABC-induced adverse reactions were observed in HLA-B*57 : 01 transgenic (B*57 : 01-Tg) mice. Moreover, regulating immune tolerance could result in severe AHS that mimics symptoms observed in the clinical setting, which were modeled in CD4+ T cell-depleted programmed death-1 receptor (PD-1) knockout B*57 : 01-Tg (B*57 : 01-Tg/PD-1-/-) mice. Here, we aimed to examine whether thymus and activation-regulated chemokine (TARC)/CCL17 level can be used as a biomarker for AHS. Serum TARC levels increased in HLA-B*57 : 01-transgenic mice following oral administration of ABC; this increase was associated with the severity of skin toxicity. In ABC-fed CD4+ T cell-depleted B*57 : 01-Tg/PD-1-/- mice, TARC was detected in the epidermal keratinocytes of the ear. Skin toxicity was characterized by the infiltration of CD8+ T cells partially expressing C-C chemokine receptor type 4, which is the primary receptor for TARC. In vivo TARC neutralization effectively alleviated the symptoms of ear skin redness and blood vessel dilatation. Moreover, TARC neutralization suppressed the infiltration of CD8+ T cells to the ear skin but did not affect the ABC-induced adaptive immune response. Therefore, TARC was involved in ABC-induced skin toxicity and contributed to the recruitment of CD8+ T cells to skin. This evidence suggests that serum TARC level may be a functional biomarker for AHS.
Assuntos
Linfócitos T CD8-Positivos , Quimiocina CCL17 , Dermatite Atópica , Animais , Linfócitos T CD8-Positivos/imunologia , Quimiocina CCL17/genética , Quimiocinas , Ciclopropanos/efeitos adversos , Didesoxiadenosina/efeitos adversos , Didesoxiadenosina/análogos & derivados , Antígenos HLA-B/genética , Humanos , Camundongos , Camundongos Transgênicos , Receptor de Morte Celular Programada 1RESUMO
Inflammatory pain, such as hypersensitivity resulting from surgical tissue injury, occurs as a result of interactions between the immune and nervous systems with the orchestrated recruitment and activation of tissue-resident and circulating immune cells to the site of injury. Our previous studies identified a central role for Ly6Clow myeloid cells in the pathogenesis of postoperative pain. We now show that the chemokines CCL17 and CCL22, with their cognate receptor CCR4, are key mediators of this response. Both chemokines are up-regulated early after tissue injury by skin-resident dendritic and Langerhans cells to act on peripheral sensory neurons that express CCR4. CCL22, and to a lesser extent CCL17, elicit acute mechanical and thermal hypersensitivity when administered subcutaneously; this response abrogated by pharmacological blockade or genetic silencing of CCR4. Electrophysiological assessment of dissociated sensory neurons from naïve and postoperative mice showed that CCL22 was able to directly activate neurons and enhance their excitability after injury. These responses were blocked using C 021 and small interfering RNA (siRNA)-targeting CCR4. Finally, our data show that acute postoperative pain is significantly reduced in mice lacking CCR4, wild-type animals treated with CCR4 antagonist/siRNA, as well as transgenic mice depleted of dendritic cells. Together, these results suggest an essential role for the peripheral CCL17/22:CCR4 axis in the genesis of inflammatory pain via direct communication between skin-resident dendritic cells and sensory neurons, opening therapeutic avenues for its control.
Assuntos
Células de Langerhans/metabolismo , Dor Pós-Operatória/etiologia , Dor Pós-Operatória/metabolismo , Receptores CCR4/metabolismo , Células Receptoras Sensoriais/metabolismo , Potenciais de Ação , Animais , Biomarcadores , Quimiocina CCL17/genética , Quimiocina CCL17/metabolismo , Quimiocina CCL22/genética , Quimiocina CCL22/metabolismo , Modelos Animais de Doenças , Suscetibilidade a Doenças , Perfilação da Expressão Gênica , Células de Langerhans/imunologia , Camundongos , Dor Pós-Operatória/diagnóstico , Transdução de SinaisRESUMO
Chemokines and adhesion molecules are major inflammatory mediators of chronic and recurrent vernal keratoconjunctivitis (VKC). Sulforaphane (SFN) is a natural plant extract that is known to have anti-inflammatory and antioxidant properties. SFN is demonstrated to be effective against a variety of human diseases. The current investigation examines the effects and the molecular mechanisms of SFN on cytokine-induced human corneal fibroblasts (HCFs) expression of adhesion molecules and chemokines. HCFs were exposed to both interleukin (IL)-4 and tumor necrosis factor (TNF)-α in the absence or presence of SFN treatment. The levels of thymus- and activation-regulated chemokine (TARC) and eotaxin-1 in culture supernatants were evaluated using enzyme-linked immunosorbent assay (ELISA). Reverse transcription-polymerase chain reaction analysis (RT-PCR) enabled quantification of mRNA levels of vascular cell adhesion molecule (VCAM)-1, eotaxin-1, and TARC along with cytokine receptors. An immunoblotting assay was used to evaluate the activities of VCAM-1, nuclear factor-kappa B (NF-κB), mitogen-activated protein kinases (MAPKs), signal transducer and activator of transcription factor (STAT)6 pathways, along with the expression of the cytokine receptors including IL-4 receptor (R)α, IL-13Rα1, TNFRI, as well as TNFRII. SFN inhibited TARC and eotaxin-1 release in HCFs stimulated by TNF-α and IL-4 in a manner dependent on dose and time. SFN suppressed transcriptions of TARC, eotaxin-1, and VCAM-1. Furthermore, the mRNA and protein expression levels of IL-4Rα, TNFRI, and TNFRII were also attenuated by SFN exposure, however, those of IL-13Rα1 remained unaffected. In addition, SFN downregulated the expression of VCAM-1 and the phosphorylation of MAPKs, IκBα, and STAT6. These results suggest that SFN inhibited cytokine-stimulated TARC, eotaxin-1 secretion as well as VCAM-1 expression in HCFs, with these effects likely occurring as a result of cytokine receptor inhibition and attenuation of MAPK, NF-κB, and STAT6 signaling. SFN may therefore have therapeutic potential in VKC treatment.
Assuntos
Quimiocinas/genética , Ceratócitos da Córnea/efeitos dos fármacos , Citocinas/antagonistas & inibidores , Isotiocianatos/farmacologia , Proteínas Quinases Ativadas por Mitógeno/metabolismo , NF-kappa B/metabolismo , Fator de Transcrição STAT1/metabolismo , Sulfóxidos/farmacologia , Molécula 1 de Adesão de Célula Vascular/genética , Anticarcinógenos/farmacologia , Sobrevivência Celular , Células Cultivadas , Quimiocina CCL11/genética , Quimiocina CCL17/genética , Ceratócitos da Córnea/metabolismo , Citocinas/farmacologia , Ensaio de Imunoadsorção Enzimática , Expressão Gênica , Regulação da Expressão Gênica/fisiologia , Humanos , Fosforilação , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Transdução de SinaisRESUMO
CCL17, a chemotactic cytokine produced by macrophages, is known to promote inflammatory and fibrotic effects in multiple organs, but its role in mediating renal fibrosis is unclear. In our study cohort of 234 chronic kidney disease (CKD) patients and 65 healthy controls, human cytokine array analysis revealed elevated CCL17 expression in CKD that correlated negatively with renal function. The area under the receiver operating characteristic curve of CCL17 to predict the development of CKD stages 3b-5 was 0.644 (p < 0.001), with the optimal cut-off value of 415.3 ng/mL. In vitro over-expression of CCL17 in HK2 cells had no effect on cell viability, but increased cell motility and the expression of α-SMA, vimentin and collagen I, as shown by western blot analysis. In a unilateral ureteral obstruction (UUO) mouse model, we observed significantly increased interstitial fibrosis and renal tubule dilatation by Masson's Trichrome and H&E staining, and markedly increased expression of CCL17, vimentin, collagen I, and α-SMA by IHC stain, qRTPCR, and western blotting. CCL17 induced renal fibrosis by promoting the epithelial-mesenchymal transition, resulting in ECM accumulation. CCL17 may be a useful biomarker for predicting the development of advanced CKD.
Assuntos
Quimiocina CCL17/metabolismo , Biomarcadores/metabolismo , Estudos de Casos e Controles , Ciclo Celular , Movimento Celular/genética , Sobrevivência Celular/genética , Quimiocina CCL17/genética , Transição Epitelial-Mesenquimal/genética , Fibrose , Taxa de Filtração Glomerular , Humanos , Rim/metabolismo , Rim/patologia , Rim/fisiopatologia , Modelos Biológicos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Insuficiência Renal Crônica/genética , Insuficiência Renal Crônica/patologia , Insuficiência Renal Crônica/fisiopatologia , Regulação para Cima/genética , Obstrução Ureteral/genética , Obstrução Ureteral/patologiaRESUMO
Tumors and infectious agents both benefit from an immunosuppressive environment. Cutibacterium acnes (C. acnes) is a bacterium in the normal skin microbiota, which has the ability to survive intracellularly in macrophages and is significantly more common in prostate cancer tissue compared with normal prostate tissue. This study investigated if prostate cancer tissue culture positive for C. acnes has a higher infiltration of regulatory T-cells (Tregs) and if macrophages stimulated with C. acnes induced the expression of immunosuppressive genes that could be linked to an increase of Tregs in prostate cancer. Real-time PCR and enzyme-linked immunosorbent spot assay (ELISA) were used to examine the expression of immunosuppressive genes in human macrophages stimulated in vitro with C. acnes, and associations between the presence of C. acnes and infiltration of Tregs were investigated by statistically analyzing data generated in two previous studies. The in vitro results demonstrated that macrophages stimulated with C. acnes significantly increased their expression of PD-L1, CCL17, and CCL18 mRNA and protein (p <0.05). In the cohort, Tregs in tumor stroma and tumor epithelia were positively associated with the presence of C. acnes (P = 0.0004 and P = 0.046, respectively). Since the macrophages stimulated with C. acnes in vitro increased the expression of immunosuppressive genes, and prostate cancer patients with prostatic C. acnes infection had higher infiltration of Tregs than their noninfected counterparts, we suggest that C. acnes may contribute to an immunosuppressive tumor environment that is vital for prostate cancer progression. IMPORTANCE In an immune suppressive tumor microenvironment constituted by immunosuppressive cells and immunosuppressive mediators, tumors may improve their ability to give rise to a clinically relevant cancer. In the present study, we found that C. acnes might contribute to an immunosuppressive environment by recruiting Tregs and by increasing the expression of immunosuppressive mediators such as PD-L1, CCL17, and CCL18. We believe that our data add support to the hypothesis of a contributing role of C. acnes in prostate cancer development. If established that C. acnes stimulates prostate cancer progression it may open up avenues for targeted prostate cancer treatment.
Assuntos
Tolerância Imunológica/imunologia , Macrófagos/imunologia , Propionibacteriaceae/imunologia , Neoplasias da Próstata/imunologia , Neoplasias da Próstata/microbiologia , Linfócitos T Reguladores/imunologia , Antígeno B7-H1/biossíntese , Antígeno B7-H1/genética , Quimiocina CCL17/biossíntese , Quimiocina CCL17/genética , Quimiocinas CC/biossíntese , Quimiocinas CC/genética , ELISPOT , Humanos , Tolerância Imunológica/genética , Masculino , Microbiota/imunologia , Neoplasias da Próstata/patologia , Evasão Tumoral/imunologia , Microambiente Tumoral/imunologiaRESUMO
An inhibitor of the histone methyltransferase enhancer of zeste homologue 2 (EZH2), tazemetostat, has been developed for the treatment of B-cell lymphoma, but its mechanisms of action are not fully elucidated. We screened for genes targeted by tazemetostat in eleven B-cell lymphoma cell lines and found that tazemetostat significantly increased the expression of chemokine (C-C motif) ligand 17 (CCL17)/thymus- and activation-regulated chemokine (TARC) in all, which codes for a chemokine that is a hallmark of Hodgkin/Reed-Sternberg (H/RS) cells in Hodgkin lymphoma. Notably, gene set enrichment analysis demonstrated a positive correlation between the genes upregulated by tazemetostat in five follicular lymphoma (FL) cell lines and those reported to be overexpressed in H/RS cells. The CCL17 promoter region was enriched in repressive histone modification H3K27me3, and tazemetostat induced H3K27 demethylation and activated gene transcription. CCL17 protein secretion was also induced by EZH2 inhibition, which was further enhanced by concurrent CpG stimulation. In vitro transwell migration assay demonstrated that CCL17 produced by tazemetostat-treated B cells enhanced the recruitment of T cells, which had the potential to exert antilymphoma response. Analysis of publicly available human lymphoma databases showed that CCL17 gene expression was inversely correlated with the EZH2 activation signature and significantly paralleled the CD4+ and CD8+ T-cell-rich signature in FL and germinal center B-cell-like diffuse large B-cell lymphoma. Our findings indicate that tazemetostat can potentially activate antilymphoma response by upregulating CCL17 expression in B-cell lymphoma cells and promote T-cell recruitment, which provides a rationale for its combination with immunotherapy.
Assuntos
Benzamidas/farmacologia , Compostos de Bifenilo/farmacologia , Quimiocina CCL17/metabolismo , Proteína Potenciadora do Homólogo 2 de Zeste/antagonistas & inibidores , Linfócitos do Interstício Tumoral , Linfoma Difuso de Grandes Células B/metabolismo , Morfolinas/farmacologia , Piridonas/farmacologia , Linhagem Celular Tumoral , Movimento Celular , Quimiocina CCL17/genética , Bases de Dados Factuais , Humanos , Histona Desmetilases com o Domínio Jumonji/genética , Linfoma Difuso de Grandes Células B/tratamento farmacológico , Linfoma Difuso de Grandes Células B/genética , Linfoma Difuso de Grandes Células B/patologia , Regiões Promotoras Genéticas , Células de Reed-Sternberg , Microambiente Tumoral/efeitos dos fármacos , Microambiente Tumoral/imunologia , Regulação para CimaRESUMO
Skullcapflavone II (SFII), a flavonoid derived from Scutellaria baicalensis, has been reported to have anti-inflammatory properties. However, its therapeutic potential for skin inflammatory diseases and its mechanism are unknown. Therefore, this study aimed to investigate the effect of SFII on TNF-α/IFN-γ-induced atopic dermatitis (AD)-associated cytokines, such as thymus- and activation-regulated chemokine (TARC) and macrophage-derived chemokine (MDC). Co-stimulation with TNF-α/IFN-γ in HaCaT cells is a well-established model for induction of pro-inflammatory cytokines. We treated cells with SFII prior to TNF-α/IFN-γ-stimulation and confirmed that it significantly inhibited TARC and MDC expression at the mRNA and protein levels. Additionally, SFII also inhibited the expression of cathepsin S (CTSS), which is associated with itching in patients with AD. Using specific inhibitors, we demonstrated that STAT1, NF-κB, and p38 MAPK mediate TNF-α/IFN-γ-induced TARC and MDC, as well as CTSS expression. Finally, we confirmed that SFII significantly suppressed TNF-α/IFN-γ-induced phosphorylation of STAT1, NF-κB, and p38 MAPK. Taken together, our study indicates that SFII inhibits TNF-α/IFN-γ-induced TARC, MDC, and CTSS expression by regulating STAT1, NF-κB, and p38 MAPK signaling pathways.
Assuntos
Catepsinas/biossíntese , Quimiocina CCL17/biossíntese , Quimiocina CCL22/biossíntese , Flavonoides/farmacologia , Interferon gama/farmacologia , Fator de Necrose Tumoral alfa/farmacologia , Catepsinas/genética , Sobrevivência Celular/efeitos dos fármacos , Quimiocina CCL17/genética , Quimiocina CCL22/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Células HaCaT , Humanos , Queratinócitos/metabolismo , NF-kappa B/metabolismo , Fator de Transcrição STAT1/metabolismo , Transdução de Sinais , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
Lupus nephritis is a severe organ manifestation in systemic lupus erythematosus leading to kidney failure in a subset of patients. In lupus-prone mice, controlled infection with Plasmodium parasites protects against the progression of autoimmune pathology including lethal glomerulonephritis. Here, we demonstrate that parasite-induced protection was not due to a systemic effect of infection on autoimmunity as previously assumed, but rather to specific alterations in immune cell infiltrates into kidneys and renal draining lymph nodes. Infection of lupus-prone mice with a Plasmodium parasite did not reduce the levels or specificities of autoreactive antibodies, vasculitis, immune complex-induced innate activation, or hypoxia. Instead, infection uniquely reduced kidney-infiltrating CCL17-producing bone marrow-derived type 2 inflammatory dendritic cells (iDC2s). Bone marrow reconstitution experiments revealed that infection with Plasmodium caused alterations in bone marrow cells that hindered the ability of DC2s to infiltrate the kidneys. The essential role for CCL17 in lupus nephritis was confirmed by in vivo depletion with a blocking antibody, which reduced kidney pathology and immune infiltrates, while bypassing the need for parasitic infection. Therefore, infiltration into the kidneys of iDC2s, with the potential to prime local adaptive responses, is an essential regulated event in the transition from manageable glomerulonephritis to lethal tubular injury.
Assuntos
Quimiocina CCL17/imunologia , Células Dendríticas/imunologia , Nefrite Lúpica/prevenção & controle , Malária/imunologia , Plasmodium yoelii/imunologia , Animais , Quimiocina CCL17/genética , Modelos Animais de Doenças , Nefrite Lúpica/genética , Nefrite Lúpica/imunologia , Malária/genética , Malária/patologia , Camundongos , Camundongos KnockoutAssuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Doença de Hodgkin/tratamento farmacológico , Doença de Hodgkin/patologia , Adolescente , Adulto , Bleomicina/uso terapêutico , Quimiocina CCL17/genética , Quimiocina CCL17/metabolismo , Ciclofosfamida/uso terapêutico , Doxorrubicina/uso terapêutico , Etoposídeo/uso terapêutico , Feminino , Perfilação da Expressão Gênica/métodos , Doença de Hodgkin/genética , Humanos , Antígeno Ki-1/genética , Antígeno Ki-1/metabolismo , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Valor Preditivo dos Testes , Prednisona/uso terapêutico , Procarbazina/uso terapêutico , Receptor alfa de Fator de Crescimento Derivado de Plaquetas/genética , Receptor alfa de Fator de Crescimento Derivado de Plaquetas/metabolismo , Taxa de Sobrevida , Transcriptoma , Resultado do Tratamento , Vincristina/uso terapêutico , Adulto JovemRESUMO
Group 2 innate lymphoid cells (ILC2s) play an important role in the pathophysiology of asthma via the robust production of type 2 cytokines. Recent studies have demonstrated that TLR7 (Toll-like receptor 7) signaling skews toward a type 1 inflammatory response in asthma, which may lead to the development of novel treatment strategies. However, the effect of TLR7 signaling on ILC2-dependent nonallergic eosinophilic inflammation remains unclear. In this study, we investigated the effects of R848, a TLR7 agonist, in a mouse model of IL-33-induced eosinophilic airway inflammation. Intranasal administration of R848 decreased infiltration of airway eosinophils and ILC2s, mucus production in epithelial cells, and type 2 cytokine production. Flow cytometric analysis identified an increased number of interstitial macrophages (IMs) expressing a high level of TLR7 in the lung upon IL-33 stimulation. IL-33-induced IMs also expressed high levels of alternatively activated (M2)-type genes and chemokines (CCL17 and CCL24). However, R848 stimulation modified these gene expressions and elicited the production of IL-27. Coculture experiments revealed that IL-33-induced IMs directly suppressed ILC2 activation in response to R848. In addition, the inhibitory effects of R848 on ILC2-induced type 2 inflammation were defective in WSX-1-deficient mice lacking the IL-27 receptor. Taken together, these findings indicate that R848 stimulates IL-33-induced IMs to suppress ILC2-mediated type 2 airway inflammation via IL-27. These findings highlight the therapeutic potential of TLR7 agonists and/or IL-27 cascades in nonallergic asthma.
Assuntos
Imidazóis/farmacologia , Imunidade Inata/efeitos dos fármacos , Interleucinas/imunologia , Pulmão/imunologia , Linfócitos/imunologia , Macrófagos/imunologia , Glicoproteínas de Membrana/agonistas , Transdução de Sinais/efeitos dos fármacos , Receptor 7 Toll-Like/agonistas , Animais , Asma/genética , Asma/imunologia , Asma/patologia , Quimiocina CCL17/genética , Quimiocina CCL17/imunologia , Quimiocina CCL24/genética , Quimiocina CCL24/imunologia , Eosinófilos/imunologia , Eosinófilos/patologia , Imunidade Inata/genética , Inflamação/genética , Inflamação/imunologia , Interleucina-33/genética , Interleucina-33/imunologia , Interleucinas/genética , Pulmão/patologia , Linfócitos/patologia , Macrófagos/patologia , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Knockout , Receptores de Interleucina/deficiência , Receptores de Interleucina/imunologia , Transdução de Sinais/genética , Transdução de Sinais/imunologia , Receptor 7 Toll-Like/genética , Receptor 7 Toll-Like/imunologiaRESUMO
Melanoma differentiation associated gene-9 (MDA-9), Syntenin-1, or syndecan binding protein is a differentially regulated prometastatic gene with elevated expression in advanced stages of melanoma. MDA-9/Syntenin expression positively associates with advanced disease stage in multiple histologically distinct cancers and negatively correlates with patient survival and response to chemotherapy. MDA-9/Syntenin is a highly conserved PDZ-domain scaffold protein, robustly expressed in a spectrum of diverse cancer cell lines and clinical samples. PDZ domains interact with a number of proteins, many of which are critical regulators of signaling cascades in cancer. Knockdown of MDA-9/Syntenin decreases cancer cell metastasis, sensitizing these cells to radiation. Genetic silencing of MDA-9/Syntenin or treatment with a pharmacological inhibitor of the PDZ1 domain, PDZ1i, also activates the immune system to kill cancer cells. Additionally, suppression of MDA-9/Syntenin deregulates myeloid-derived suppressor cell differentiation via the STAT3/interleukin (IL)-1ß pathway, which concomitantly promotes activation of cytotoxic T lymphocytes. Biologically, PDZ1i treatment decreases metastatic nodule formation in the lungs, resulting in significantly fewer invasive cancer cells. In summary, our observations indicate that MDA-9/Syntenin provides a direct therapeutic target for mitigating aggressive breast cancer and a small-molecule inhibitor, PDZ1i, provides a promising reagent for inhibiting advanced breast cancer pathogenesis.
Assuntos
Antineoplásicos/farmacologia , Neoplasias da Mama/tratamento farmacológico , Interleucina-1beta/genética , Neoplasias Pulmonares/tratamento farmacológico , Oxidiazóis/farmacologia , Pirimidinas/farmacologia , Sinteninas/genética , Animais , Antineoplásicos/síntese química , Neoplasias da Mama/genética , Neoplasias da Mama/imunologia , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Quimiocina CCL11/genética , Quimiocina CCL11/imunologia , Quimiocina CCL17/genética , Quimiocina CCL17/imunologia , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Interleucina-10/genética , Interleucina-10/imunologia , Interleucina-1alfa/genética , Interleucina-1alfa/imunologia , Interleucina-1beta/antagonistas & inibidores , Interleucina-1beta/imunologia , Subunidade p19 da Interleucina-23/genética , Subunidade p19 da Interleucina-23/imunologia , Interleucina-5/genética , Interleucina-5/imunologia , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/imunologia , Neoplasias Pulmonares/secundário , Camundongos , Camundongos Endogâmicos BALB C , Oxidiazóis/síntese química , Pirimidinas/síntese química , Transdução de Sinais , Sinteninas/antagonistas & inibidores , Sinteninas/imunologia , Linfócitos T Citotóxicos/efeitos dos fármacos , Linfócitos T Citotóxicos/imunologia , Linfócitos T Citotóxicos/patologia , Carga Tumoral/efeitos dos fármacos , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
3,4,5Trihydroxycinnamic acid (THCA) exhibits antiinflammatory activity in acute or chronic inflammatory disorders, such as acute lung injury and asthma. The present study investigated the antiinflammatory activity of THCA in a tumor necrosis factorα/interferonγ (TI) mixturestimulated human keratinocyte cell line. The results of ELISA and reverse transcriptionquantitative PCR revealed that THCA reduced the secretion and mRNA expression levels of interleukin (IL)6; IL8; thymus and activationregulated chemokine; macrophagederived chemokine; regulated upon activation, normal T cell expressed and secreted; and monocyte chemoattractant protein1 in TI mixturestimulated HaCaT cells. In addition, the results of western blot analysis demonstrated that THCA exerted inhibitory activity on the activation of AKT, ERK and nuclear factorκB in TI mixturestimulated HaCaT cells. Furthermore, THCA upregulated the expression levels of heme oxygenase1 and NAD(P)H:quinone oxidoreductase 1, and the activation of nuclear factor erythroid 2related factor 2 in HaCaT cells. These results demonstrated that THCA may exhibit antiinflammatory activity in activated HaCaT cells.