RESUMEN
Long non-coding RNA (lncRNA) LINC00472 has a close connection with the development of tumors. The aim was to explore the role of LINC00472 on NSCLC cell biological function in vivo and its potential mechanisms. The mRNA levels of LncRNA 00472 and microRNA-23a-3p, were determined by RT-qPCR. Cell Counting Kit-8, cell scratches and western blot assays were used to analyze the proliferation, migration and level of apoptosis-associated proteins. Luciferase reporter assay validates the binding between LINC00472/CCL22 and miR-23a-3p. LINC00472 and CCL22 were lowly expressed in NSCLC tissues and cells, while miR-23a-3p expression was upregulated. LINC00472 overexpression significantly depressed NSCLC cell cellular behavior, whereas promoting cell death. MiR-23a-3p could reverse these above-mentioned biological behavior changes caused by LINC00472 overexpression. Additionally, LINC00472 increased CCL22 expression through sponging miR-23a-3p. Knocking down CCL22 antagonized the inhibitory effect of LINC00472 on NSCLC cell survival. LINC00472 may reduce the cellular growth, and accelerate death of NSCLC through increasing CCL22 expression by targeting miR-23a-3p.
Asunto(s)
Apoptosis , Carcinoma de Pulmón de Células no Pequeñas , Movimiento Celular , Proliferación Celular , Quimiocina CCL22 , Regulación Neoplásica de la Expresión Génica , Neoplasias Pulmonares , MicroARNs , ARN Largo no Codificante , Carcinoma de Pulmón de Células no Pequeñas/genética , Carcinoma de Pulmón de Células no Pequeñas/patología , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Humanos , MicroARNs/genética , MicroARNs/metabolismo , ARN Largo no Codificante/genética , ARN Largo no Codificante/metabolismo , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patología , Neoplasias Pulmonares/metabolismo , Proliferación Celular/genética , Línea Celular Tumoral , Quimiocina CCL22/genética , Quimiocina CCL22/metabolismo , Apoptosis/genética , Movimiento Celular/genética , Progresión de la Enfermedad , Masculino , Femenino , AnimalesRESUMEN
CCL22 is a macrophage-derived immunosuppressive chemokine that recruits regulatory T cells through the CCL22:CCR4 axis, playing an important role in homeostatic and inflammatory responses. A CCL22-overexpressing human induced pluripotent stem cell line (CNNDi001-A-2) was generated by lentiviral transduction to further study the function of CCL22. The cell line was confirmed to have normal proliferation and pluripotency and could be further differentiated into islet cells for cell replacement therapy in diabetes.
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Células Madre Pluripotentes Inducidas , Islotes Pancreáticos , Humanos , Células Madre Pluripotentes Inducidas/metabolismo , Quimiocina CCL22/genética , Quimiocina CCL22/metabolismo , Línea Celular , Linfocitos T Reguladores/metabolismoRESUMEN
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.
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Células Endoteliales , Obesidad Mórbida , Humanos , Adulto , Persona de Mediana Edad , Células Endoteliales/metabolismo , Obesidad Mórbida/complicaciones , Obesidad Mórbida/cirugía , Quimiocina CCL17/genética , Quimiocinas , Transducción de Señal , Receptores de Quimiocina/metabolismo , Quimiocina CCL22/genéticaRESUMEN
OBJECTIVES: Chemokines have been suggested to play significant roles in the progression of malignant cancers. This study aimed to identify the chemokines related to malignant progression in thyroid carcinoma. METHODS: The mRNA expression levels of 52 chemokines were compared between differentiated thyroid cancer (DTC) samples and normal thyroid tissues from The Cancer Genome Atlas database; survival analysis was then performed on the basis of differentially expressed chemokines. A retrospective study was conducted on the level of differentially expressed chemokines in 76 DTC patients. Functional pathway analysis was performed to explore chemokine-related regulatory mechanisms. RESULTS: We identified 20 chemokines with differentially expressed mRNA levels through publicly available data. High levels of CCL22 and CCL26 were found to be related with metastasis in clinical DTC samples. High levels of CCL22 were found to be significantly related to poor prognosis in DTC patients. Pathway analyses revealed that cytokines might affect cancer progression through cytokine-cytokine receptor and cytokine-interleukin interactions. CONCLUSIONS: CCL22 and CCL26 could serve as prognostic biomarkers in thyroid carcinoma.
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Adenocarcinoma , Neoplasias de la Tiroides , Humanos , Estudios Retrospectivos , Neoplasias de la Tiroides/patología , Biomarcadores , Adenocarcinoma/secundario , Quimiocinas/genética , ARN Mensajero , Pronóstico , Quimiocina CCL22/genética , Quimiocina CCL26RESUMEN
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.
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Linfadenopatía Inmunoblástica , Linfoma de Células T , Quimiocina CCL17/genética , Quimiocina CCL17/uso terapéutico , Quimiocina CCL22/genética , Quimiocina CCL22/uso terapéutico , Ciclofosfamida/uso terapéutico , Doxorrubicina/uso terapéutico , Humanos , Linfadenopatía Inmunoblástica/tratamiento farmacológico , Linfadenopatía Inmunoblástica/genética , Linfadenopatía Inmunoblástica/patología , Linfoma de Células T/tratamiento farmacológico , Linfoma de Células T/genética , Linfoma de Células T/patología , Prednisona/uso terapéutico , Transcriptoma , Microambiente Tumoral/genética , Vincristina/uso terapéuticoRESUMEN
Chronic lymphoproliferative disorder of natural killer cells (CLPD-NK) is characterized by clonal expansion of natural killer (NK) cells where the underlying genetic mechanisms are incompletely understood. In the present study, we report somatic mutations in the chemokine gene CCL22 as the hallmark of a distinct subset of CLPD-NK. CCL22 mutations were enriched at highly conserved residues, mutually exclusive of STAT3 mutations and associated with gene expression programs that resembled normal CD16dim/CD56bright NK cells. Mechanistically, the mutations resulted in ligand-biased chemokine receptor signaling, with decreased internalization of the G-protein-coupled receptor (GPCR) for CCL22, CCR4, via impaired ß-arrestin recruitment. This resulted in increased cell chemotaxis in vitro, bidirectional crosstalk with the hematopoietic microenvironment and enhanced NK cell proliferation in vivo in transgenic human IL-15 mice. Somatic CCL22 mutations illustrate a unique mechanism of tumor formation in which gain-of-function chemokine mutations promote tumorigenesis by biased GPCR signaling and dysregulation of microenvironmental crosstalk.
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Quimiocina CCL22 , Células Asesinas Naturales , Trastornos Linfoproliferativos , Animales , Quimiocina CCL22/genética , Células Asesinas Naturales/patología , Activación de Linfocitos , Trastornos Linfoproliferativos/genética , Trastornos Linfoproliferativos/metabolismo , Trastornos Linfoproliferativos/patología , Ratones , MutaciónRESUMEN
Somatic mutations in CCL22 were found in a distinct subset of CLPD-NK and drive disease development.
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Trastornos Linfoproliferativos , Quimiocina CCL22/genética , Enfermedad Crónica , Humanos , Células Asesinas Naturales , Trastornos Linfoproliferativos/genética , MutaciónRESUMEN
Long noncoding RNA (LncRNA) is a new type of regulatory RNA. LncRNA HOX antisense intergenic RNA (HOTAIR), as an oncogene in non-small cell lung cancer (NSCLC), is one of the key determinants of tumor progression. However, its possible molecular mechanism and the immunomodulatory pathway involved in NSCLC are still unclear. This study aims to explore whether HOTAIR promotes proliferation, migration and invasion of the NSCLC cells by inhibiting the expression of C-C Motif Chemokine Ligand 22 (CCL22). We collected 30 clinical samples of cancer and adjacent normal tissues from the patients with NSCLC, using real-time quantitative polymerase chain reaction (RT-qPCR) to detect the LncRNA HOTAIR and CCL22 mRNA expression in tissues. Immunohistochemistry was used to detect the protein expression of CCL22 in cancer and adjacent normal tissues. Cell experiments were conducted to verify that LncRNA HOTAIR regulates the expression of CCL22 and participates in the progress of NSCLC. The antisense oligonucleotide (ASO) probe interfering with LncRNA HOTAIR and the interference fragment of CCL22 (si-CCL22) were constructed. A549 cells were co-transfected with ASO-HOTAIR and si-CCL22. We used RT-qPCR to detect the expression of LncRNA HOTAIR and CCL22 mRNA in the cells, enzyme-linked immunosorbent assay (ELISA) used to detect the CCL22 protein level in the cell supernatant. 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was applied to detect cell proliferation, the Flow cytometry to detect cell apoptosis. Finally, the Transwell test was utilized to detect cell migration and invasion. In conclusion, this study suggests that HOTAIR may promote proliferation, migration and invasion of the NSCLC cells by inhibiting CCL22 expression, which may play a key role in NSCLC cell immunity.
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Biomarcadores de Tumor/metabolismo , Carcinoma de Pulmón de Células no Pequeñas/patología , Movimiento Celular , Proliferación Celular , Quimiocina CCL22/metabolismo , Regulación Neoplásica de la Expresión Génica , ARN Largo no Codificante/genética , Apoptosis , Biomarcadores de Tumor/genética , Carcinoma de Pulmón de Células no Pequeñas/genética , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Quimiocina CCL22/genética , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , Invasividad Neoplásica , Pronóstico , Células Tumorales CultivadasRESUMEN
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.
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Células de Langerhans/metabolismo , Dolor Postoperatorio/etiología , Dolor Postoperatorio/metabolismo , Receptores CCR4/metabolismo , Células Receptoras Sensoriales/metabolismo , Potenciales de Acción , Animales , Biomarcadores , Quimiocina CCL17/genética , Quimiocina CCL17/metabolismo , Quimiocina CCL22/genética , Quimiocina CCL22/metabolismo , Modelos Animales de Enfermedad , Susceptibilidad a Enfermedades , Perfilación de la Expresión Génica , Células de Langerhans/inmunología , Ratones , Dolor Postoperatorio/diagnóstico , Transducción de SeñalRESUMEN
BACKGROUND: Asthma is a heterogeneous disease and different phenotypes based on clinical parameters have been identified. However, the molecular subgroups of asthma defined by gene expression profiles of induced sputum have been rarely reported. METHODS: We re-analyzed the asthma transcriptional profiles of the dataset of GSE45111. A deep bioinformatics analysis was performed. We classified 47 asthma cases into different subgroups using unsupervised consensus clustering analysis. Clinical features of the subgroups were characterized, and their biological function and immune status were analyzed using Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) and single sample Gene Set Enrichment Analysis (ssGSEA). Weighted gene co-expression network analysis (WGCNA) and protein-protein interaction (PPI) network were performed to identify key gene modules and hub genes. RESULTS: Unsupervised consensus clustering of gene expression profiles in asthma identified two distinct subgroups (Cluster I/II), which were significantly associated with eosinophilic asthma (EA) and paucigranulocytic asthma (PGA). The differentially expressed genes (DEGs) between the two subgroups were primarily enriched in immune response regulation and signal transduction. The ssGSEA suggested the different immune infiltration and function scores between the two clusters. The WGCNA and PPI analysis identified three hub genes: THBS1, CCL22 and CCR7. ROC analysis further suggested that the three hub genes had a good ability to differentiate the Cluster I from the Cluster II. CONCLUSIONS: Based on the gene expression profiles of the induced sputum, we identified two asthma subgroups, which revealed different clinical characteristics, gene expression patterns, biological functions and immune status. The transcriptional classification confirms the molecular heterogeneity of asthma and provides a framework for more in-depth research on the mechanisms of asthma.
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Asma/genética , Quimiocina CCL22/genética , Análisis por Conglomerados , Bases de Datos Genéticas , Perfilación de la Expresión Génica , Humanos , Inflamación/genética , Receptores CCR7/genética , Trombospondina 1/genéticaRESUMEN
Toxoplasma gondii is an intracellular protozoan pathogen of humans that can cross the placenta and result in adverse pregnancy outcomes and long-term birth defects. The mechanisms used by T. gondii to cross the placenta are unknown, but complex interactions with the host immune response are likely to play a role in dictating infection outcomes during pregnancy. Prior work showed that T. gondii infection dramatically and specifically increases the secretion of the immunomodulatory chemokine CCL22 in human placental cells during infection. Given the important role of this chemokine during pregnancy, we hypothesized that CCL22 induction was driven by a specific T. gondii-secreted effector. Using a combination of bioinformatics and molecular genetics, we have now identified T. gondii GRA28 as the gene product required for CCL22 induction. GRA28 is secreted into the host cell, where it localizes to the nucleus, and deletion of the GRA28 gene results in reduced CCL22 placental cells as well as a human monocyte cell line. The impact of GRA28 on CCL22 production is also conserved in mouse immune and placental cells both in vitro and in vivo. Moreover, parasites lacking GRA28 are impaired in their ability to disseminate throughout the animal, suggesting a link between CCL22 induction and the ability of the parasite to cause disease. Overall, these data demonstrate a clear function for GRA28 in altering the immunomodulatory landscape during infection of both placental and peripheral immune cells and show a clear impact of this immunomodulation on infection outcome. IMPORTANCE Toxoplasma gondii is a globally ubiquitous pathogen that can cause severe disease in HIV/AIDS patients and can also cross the placenta and infect the developing fetus. We have found that placental and immune cells infected with T. gondii secrete significant amounts of a chemokine (called CCL22) that is critical for immune tolerance during pregnancy. In order to better understand whether this is a response by the host or a process that is driven by the parasite, we have identified a T. gondii gene that is absolutely required to induce CCL22 production in human cells, indicating that CCL22 production is a process driven almost entirely by the parasite rather than the host. Consistent with its role in immune tolerance, we also found that T. gondii parasites lacking this gene are less able to proliferate and disseminate throughout the host. Taken together, these data illustrate a direct relationship between CCL22 levels in the infected host and a key parasite effector and provide an interesting example of how T. gondii can directly modulate host signaling pathways in order to facilitate its growth and dissemination.
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Quimiocina CCL22/metabolismo , Placenta/parasitología , Complicaciones Parasitarias del Embarazo/metabolismo , Proteínas Protozoarias/metabolismo , Toxoplasma/metabolismo , Toxoplasmosis/metabolismo , Animales , Quimiocina CCL22/genética , Femenino , Interacciones Huésped-Parásitos , Humanos , Ratones , Ratones Endogámicos BALB C , Placenta/metabolismo , Embarazo , Complicaciones Parasitarias del Embarazo/genética , Complicaciones Parasitarias del Embarazo/parasitología , Proteínas Protozoarias/genética , Toxoplasma/genética , Toxoplasmosis/genética , Toxoplasmosis/parasitologíaRESUMEN
OBJECTIVE: Tongue and mouth floor squamous cell carcinoma (T/MF SCC) exhibits a high rate of local recurrence and cervical lymph node metastasis. The effect of the tumor microenvironment on T/MF SCC remains unclear. MATERIALS AND METHODS: Transcriptome and somatic mutation data of patients with T/MF SCC were obtained from HNSC projects of the Cancer Genome Atlas. Immune infiltration quantification in early- (clinical stage I-II) and advanced-stage (clinical stage III-IV) T/MF SCC was performed using single sample Gene Set Enrichment Analysis and MCPcounter. Differentially expressed gene data were filtered, and their function was assessed through Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses. Kaplan-Meier survival curve analysis and Cox regression model were conducted to evaluate the survival of patients with the CCL22 signature. Maftools was used to present the overview of somatic mutations. RESULTS: In T/MF SCC, T helper (Th)2 cell counts were significantly increased in patients with early-stage disease compared to those with advanced-stage disease. Expression of the Th2 cell-related chemokine, CCL22, was downregulated in patients with advanced-stage T/MF SCC. Univariate and multivariate Cox analyses revealed that CCL22 was a good prognostic factor in T/MF SCC. A nomogram based on the expression of CCL22 was constructed to serve as a prognostic indicator for T/MF SCC. NOTCH1 mutations were found at a higher rate in patients with advanced-stage T/MF SCC than in those with early-stage T/MF SCC, resulting in the inhibition of the activation of the NOTCH1-Th2 cell differentiation pathway. The expression levels of CCL22, GATA-3, and IL4 were higher in patients with early-stage T/MF SCC than in those with advanced-stage T/MF SCC. CONCLUSION: In T/MF SCC, high expression of CCL22 may promote the recruitment of Th2 cells and help predict a better survival. Mutations in NOTCH1 inhibit the differentiation of Th2 cells, facilitating tumor progression through a decrease in Th2 cell recruitment and differentiation.
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Carcinoma de Células Escamosas/etiología , Carcinoma de Células Escamosas/metabolismo , Quimiocina CCL22/genética , Neoplasias de la Boca/etiología , Neoplasias de la Boca/metabolismo , Receptor Notch1/genética , Células Th2/inmunología , Células Th2/metabolismo , Adulto , Anciano , Carcinoma de Células Escamosas/mortalidad , Carcinoma de Células Escamosas/patología , Quimiotaxis de Leucocito/genética , Quimiotaxis de Leucocito/inmunología , Biología Computacional/métodos , Femenino , Factor de Transcripción GATA3/genética , Factor de Transcripción GATA3/metabolismo , Perfilación de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Humanos , Estimación de Kaplan-Meier , Recuento de Linfocitos , Linfocitos Infiltrantes de Tumor , Masculino , Persona de Mediana Edad , Suelo de la Boca/metabolismo , Suelo de la Boca/patología , Neoplasias de la Boca/mortalidad , Neoplasias de la Boca/patología , Mutación , Estadificación de Neoplasias , Pronóstico , Modelos de Riesgos ProporcionalesRESUMEN
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.
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Catepsinas/biosíntesis , Quimiocina CCL17/biosíntesis , Quimiocina CCL22/biosíntesis , Flavonoides/farmacología , Interferón gamma/farmacología , Factor de Necrosis Tumoral alfa/farmacología , Catepsinas/genética , Supervivencia Celular/efectos de los fármacos , Quimiocina CCL17/genética , Quimiocina CCL22/genética , Regulación de la Expresión Génica/efectos de los fármacos , Células HaCaT , Humanos , Queratinocitos/metabolismo , FN-kappa B/metabolismo , Factor de Transcripción STAT1/metabolismo , Transducción de Señal , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
BACKGROUND: In Mali, cutaneous leishmaniasis (CL) and filariasis are co-endemic. Previous studies in animal models of infection have shown that sand fly saliva enhance infectivity of Leishmania parasites in naïve hosts while saliva-specific adaptive immune responses may protect against cutaneous and visceral leishmaniasis. In contrast, the human immune response to Phlebotomus duboscqi (Pd) saliva, the principal sand fly vector in Mali, was found to be dichotomously polarized with some individuals having a Th1-dominated response and others having a Th2-biased response. We hypothesized that co-infection with filarial parasites may be an underlying factor that modulates the immune response to Pd saliva in endemic regions. METHODOLOGY/PRINCIPAL FINDINGS: To understand which cell types may be responsible for polarizing human responses to sand fly saliva, we investigated the effect of salivary glands (SG) of Pd on human monocytes. To this end, elutriated monocytes were cultured in vitro, alone, or with SG, microfilariae antigen (MF ag) of Brugia malayi, or LPS, a positive control. The mRNA expression of genes involved in inflammatory or regulatory responses was then measured as were cytokines and chemokines associated with these responses. Monocytes of individuals who were not exposed to sand fly bites (mainly North American controls) significantly upregulated the production of IL-6 and CCL4; cytokines that enhance leishmania parasite establishment, in response to SG from Pd or other vector species. This selective inflammatory response was lost in individuals that were exposed to sand fly bites which was not changed by co-infection with filarial parasites. Furthermore, infection with filarial parasites resulted in upregulation of CCL22, a type-2 associated chemokine, both at the mRNA levels and by its observed effect on the frequency of recruited monocytes. CONCLUSIONS/SIGNIFICANCE: Together, our data suggest that SG or recombinant salivary proteins from Pd alter human monocyte function by upregulating selective inflammatory cytokines.
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Brugia Malayi/inmunología , Proteínas de Insectos/inmunología , Monocitos/parasitología , Phlebotomus/inmunología , Saliva/inmunología , Inmunidad Adaptativa , Animales , Células Cultivadas , Quimiocina CCL22/genética , Quimiocina CCL22/metabolismo , Coinfección , Enfermedades Endémicas , Filariasis/complicaciones , Filariasis/inmunología , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/inmunología , Inmunidad Celular , Leishmaniasis Cutánea/complicaciones , Leishmaniasis Cutánea/inmunología , Lipopolisacáridos/toxicidad , Malí , Monocitos/fisiología , ARN Mensajero , Proteínas Recombinantes , Glándulas Salivales , Linfocitos T Colaboradores-InductoresRESUMEN
Atopic dermatitis (AD) is a chronic inflammatory skin disease that is characterized by an impaired skin barrier and intense itchiness, which decreases the individual's quality of life. No fully effective therapeutic agents have prevailed for AD due to an insufficient grasp of the complex etiology. Ellagic acid (EA), a natural compound, has anti-inflammatory properties in chronic diseases. The effects of EA on AD have not yet been explored. The present study investigated the effects of EA on TNF-α/IFN-γ-stimulated HaCaT keratinocytes and house dust mite-induced AD-like skin lesions in NC/Nga mice. Treatment with EA suppressed inflammatory responses in keratinocytes by regulating critical inflammatory signaling pathways, such as mitogen-activated protein kinases and signal transducers and activators of transcription. In vivo studies using a DfE-induced AD mouse model showed the effects of EA administration through ameliorated skin lesions via decremented histological inflammatory reactions. These results suggest that EA could be a potential therapeutic alternative for the treatment of AD by inhibiting inflammatory signaling pathways.
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Antiinflamatorios/farmacología , Dermatitis Atópica/tratamiento farmacológico , Dermatophagoides farinae/química , Ácido Elágico/farmacología , Proteínas Quinasas Activadas por Mitógenos/genética , Factor de Transcripción STAT1/genética , Factor de Transcripción STAT3/genética , Animales , Antígenos Dermatofagoides/administración & dosificación , Quimiocina CCL17/genética , Quimiocina CCL17/inmunología , Quimiocina CCL22/genética , Quimiocina CCL22/inmunología , Quimiocina CCL5/genética , Quimiocina CCL5/inmunología , Mezclas Complejas/administración & dosificación , Citocinas/genética , Citocinas/inmunología , Dermatitis Atópica/inducido químicamente , Dermatitis Atópica/genética , Dermatitis Atópica/inmunología , Dermatophagoides farinae/inmunología , Modelos Animales de Enfermedad , Femenino , Regulación de la Expresión Génica , Células HaCaT , Humanos , Interferón gamma/antagonistas & inhibidores , Interferón gamma/farmacología , Ratones , Proteínas Quinasas Activadas por Mitógenos/inmunología , Factor de Transcripción STAT1/inmunología , Factor de Transcripción STAT3/inmunología , Transducción de Señal , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Factor de Necrosis Tumoral alfa/farmacología , Linfopoyetina del Estroma TímicoRESUMEN
Antibody affinity maturation depends on positive selection in germinal centres (GCs) of rare B cell clones that acquire higher-affinity B cell receptors via somatic hypermutation, present more antigen to follicular helper T (TFH) cells and, consequently, receive more contact-dependent T cell help1. As these GC B cells and TFH cells do not maintain long-lasting contacts in the chaotic GC environment2-4, it is unclear how sufficient T cell help is cumulatively focused onto those rare clones. Here we show that, upon stimulation of CD40, GC B cells upregulate the chemokine CCL22 and to a lesser extent CCL17. By engaging the chemokine receptor CCR4 on TFH cells, CCL22 and CCL17 can attract multiple helper cells from a distance, thus increasing the chance of productive help. During a GC response, B cells that acquire higher antigen-binding affinities express higher levels of CCL22, which in turn 'highlight' these high-affinity GC B cells. Acute increase or blockade of TFH cells helps to rapidly increase or decrease CCL22 expression by GC B cells, respectively. Therefore, a chemokine-based intercellular reaction circuit links the amount of T cell help that individual B cells have received recently to their subsequent ability to attract more help. When CCL22 and CCL17 are ablated in B cells, GCs form but B cells are not affinity-matured efficiently. When competing with wild-type B cells in the same reaction, B cells lacking CCL22 and CCL17 receive less T cell help to maintain GC participation or develop into bone-marrow plasma cells. By uncovering a chemokine-mediated mechanism that highlights affinity-improved B cells for preferential help from TFH cells, our study reveals a principle of spatiotemporal orchestration of GC positive selection.
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Quimiocina CCL22/metabolismo , Centro Germinal/citología , Centro Germinal/inmunología , Animales , Linfocitos B/citología , Linfocitos B/inmunología , Células Cultivadas , Quimiocina CCL17/deficiencia , Quimiocina CCL17/genética , Quimiocina CCL22/deficiencia , Quimiocina CCL22/genética , Femenino , Humanos , Masculino , Ratones , Tonsila Palatina/citología , Receptores CCR4/deficiencia , Receptores CCR4/genética , Linfocitos T Colaboradores-Inductores/citología , Linfocitos T Colaboradores-Inductores/inmunología , Regulación hacia ArribaRESUMEN
CCL22 is a key mediator of leukocyte trafficking in inflammatory immune responses, allergy, and cancer. It acts by attracting regulatory T cells and Th2 cells via their receptor CCR type 4 (CCR4). Beyond its role in inflammation, CCL22 is constitutively expressed at high levels in lymphoid organs during homeostasis, where it controls immunity by recruiting regulatory T cells to dendritic cells (DCs). In this study, we aimed to identify the mechanisms responsible for constitutive CCL22 expression. We confirmed that CD11c+ DCs are the exclusive producers of CCL22 in secondary lymphatic organs during homeostasis. We show that in vitro both murine splenocytes and human PBMCs secrete CCL22 spontaneously without any further stimulation. Interestingly, isolated DCs alone, however, are unable to produce CCL22, but instead require T cell help. In vitro, only the coculture of DCs with T cells or their supernatants resulted in CCL22 secretion, and we identified T cell-derived GM-CSF as the major inducer of DC-derived CCL22 expression. In vivo, Rag1 -/- mice, which lack functional T cells, have low CCL22 levels in lymphoid organs, and this can be restored by adoptive transfer of wild-type T cells or administration of GM-CSF. Taken together, we uncover T cell-derived GM-CSF as a key inducer of the chemokine CCL22 and thus, to our knowledge, identify a novel role for this cytokine as a central regulator of immunity in lymphatic organs. This knowledge could contribute to the development of new therapeutic interventions in cancer and autoimmunity.
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Quimiocina CCL22/inmunología , Células Dendríticas/inmunología , Regulación de la Expresión Génica/inmunología , Factor Estimulante de Colonias de Granulocitos y Macrófagos/inmunología , Linfocitos T Reguladores/inmunología , Animales , Antígenos CD11/genética , Antígenos CD11/inmunología , Quimiocina CCL22/genética , Células Dendríticas/citología , Hormona Liberadora de Gonadotropina/análogos & derivados , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Ratones Noqueados , Linfocitos T Reguladores/citología , Células Th2/citología , Células Th2/inmunologíaRESUMEN
PURPOSE: Radiation therapy elicits profound alterations in gene expression in tumor cells. This study aims to determine the dynamic changes in the expression of immunity-associated genes in nasopharyngeal carcinoma (NPC) cells upon radiation therapy. METHODS AND MATERIALS: The study was performed using NPC patient-derived tumor xenograft tumors, cell lines, CCR4+ CD8 T cells sorted from peripheral blood mononuclear cells of healthy volunteers, and TCGA-derived bulk RNA-seq or single-cell RNA-seq (scRNA-seq) data sets. Patient-derived tumor xenograft tumors or cell lines were irradiated and collected for bulk RNA sequencing or for CCL22 expression and release detection. Malignant phenotypes and radiosensitivity were assessed in cells with or without overexpression of CCL22 or recombinant CCL22 treatment in the presence or absence of irradiation. TCGA data sets were used for uncovering CCR4 status in subtypes of T cells. CCL22 in supernatants, cell lysates, or serum samples was measured with enzyme-linked immunosorbent assay. RESULTS: CCL22 was significantly increased in the irradiated patient-derived tumor xenograft tumors, the supernatants and cell lysates collected from irradiated NPC cell lines, and the serum of patients who received radiation therapy. No alterations of malignant phenotypes were found in tumor cells with CCL22 overexpression or recombinant CCL22 treatment. Kaplan-Meier analysis revealed that CCL22 or its receptor CCR4 positively correlated with cytotoxic T lymphocyte signatures, and high expression of CCL22 or CCR4 was associated with better prognosis for patients with NPC. scRNA-seq data set-based analysis demonstrated that CCR4 was expressed in multiple subtypes of T cells, including effector CD8 T cells. Chemotaxis assay indicated that CCR4+ CD8 T cells could be recruited by CCL22 treatment. CONCLUSION: The radiation-enhanced release of CCL22 from NPC cells promotes migration of CCR4 + effector CD8 T cells, which might partially be associated with radiation therapy-mediated antitumor immunity.
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Linfocitos T CD8-positivos/inmunología , Quimiocina CCL22/genética , Regulación Neoplásica de la Expresión Génica/efectos de la radiación , Carcinoma Nasofaríngeo/inmunología , Carcinoma Nasofaríngeo/radioterapia , Receptores CCR4/metabolismo , Animales , Linfocitos T CD8-positivos/citología , Linfocitos T CD8-positivos/metabolismo , Línea Celular Tumoral , Transformación Celular Neoplásica , Humanos , Ratones , Carcinoma Nasofaríngeo/genéticaRESUMEN
BACKGROUND: Macrophages (MÑ) can be M1/M2 polarized by Th1/2 signals, respectively. M2-like MÑ are thought to be important in asthma pathogenesis, and M1-like in anti-infective immunity, however their roles in virus-induced asthma exacerbations are unknown. Our objectives were (i) to assess polarised MÑ phenotype responses to rhinovirus (RV) infection in vitro and (ii) to assess MÑ phenotypes in healthy subjects and people with asthma before and during experimental RV infection in vivo. METHODS: We investigated characteristics of polarized/unpolarized human monocyte-derived MÑ (MDM, from 3-6 independent donors) in vitro and evaluated frequencies of M1/M2-like bronchoalveolar lavage (BAL) MÑ in experimental RV-induced asthma exacerbation in 7 healthy controls and 17 (at baseline) and 18 (at day 4 post infection) people with asthma. FINDINGS: We observed in vitro: M1-like but not M2-like or unpolarized MDM are potent producers of type I and III interferons in response to RV infection (P<0.0001), and M1-like are more resistant to RV infection (P<0.05); compared to M1-like, M2-like MDM constitutively produced higher levels of CCL22/MDC (P = 0.007) and CCL17/TARC (P<0.0001); RV-infected M1-like MDM were characterized as CD14+CD80+CD197+ (P = 0.002 vs M2-like, P<0.0001 vs unpolarized MDM). In vivo we found reduced percentages of M1-like CD14+CD80+CD197+ BAL MÑ in asthma during experimental RV16 infection compared to baseline (P = 0.024). INTERPRETATION: Human M1-like BAL MÑ are likely important contributors to anti-viral immunity and their numbers are reduced in patients with allergic asthma during RV-induced asthma exacerbations. This mechanism may be one explanation why RV-triggered clinical and pathologic outcomes are more severe in allergic patients than in healthy subjects. FUNDING: ERC FP7 Advanced grant 233015, MRC Centre Grant G1000758, Asthma UK grant 08-048, NIHR Biomedical Research Centre funding scheme, NIHR BRC Centre grant P26095, the Predicta FP7 Collaborative Project grant 260895, RSF grant 19-15-00272, Megagrant No 14.W03.31.0024.
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Asma/inmunología , Interferones/genética , Macrófagos Alveolares/inmunología , Infecciones por Picornaviridae/complicaciones , Asma/etiología , Asma/virología , Células Cultivadas , Quimiocina CCL17/genética , Quimiocina CCL17/metabolismo , Quimiocina CCL22/genética , Quimiocina CCL22/metabolismo , Células HeLa , Humanos , Interferones/metabolismo , Macrófagos Alveolares/virología , Infecciones por Picornaviridae/inmunología , Receptores Inmunológicos/genética , Receptores Inmunológicos/metabolismoRESUMEN
The HBV-initiated hepatocellular carcinoma (HCC) frequently develops from or accompanies long-term chronic hepatitis, inflammation, and cirrhosis, and has a poor prognosis. Sorafenib, an orally active multi-kinase inhibitor, currently the most common approved drug for first-line systemic treatment of advanced HCC, only improves overall survival of three months, suggesting the need for new therapeutic strategies. In this study, we identified that sorafenib selectively resisted in immune competent C57BL/6 mice but not nude mice. The chemokines CCL22 and CCL17 were upregulated by sorafenib, which elevated dramatically higher in HBV-associated HCC. Mechanically, sorafenib accelerates CCL22 expression via TNF-α-RIP1-NF-κB signaling pathway. Blocking CCL22 signaling with antagonist C-021 and sorafenib treated in combination can inhibit tumor growth and enhance the antitumor response, whereas no significant differences in tumor burden were observed in nude mice upon addition of C-021. These findings strongly suggest that CCL22 signaling pathway strongly contributes to sorafenib resistance in HBV-associated HCC, indicating a potential therapeutic strategy for immunological chemotherapy complementing first-line agents against HBV-associated HCC.