RESUMEN
Pleural fluid (PF) immune response in anergic tuberculous pleural effusion (TPE) patients is poorly understood. This study aimed to compare PF biochemical parameters and chemokine levels between anergic and non-anergic TPE patients. Chemokine arrays, cytokine measurements, and flow cytometry were performed in 58 patients (TPE [non-anergic (n = 32) and anergic (n = 10)] and malignant pleural effusion (MPE) [n = 16]). PF adenosine deaminase 2 (ADA2) levels were significantly lower in anergic TPE patients than in non-anergic TPE patients (p = 0.048). Among the 40 chemokines tested, PF CCL27 levels were significantly higher in anergic TPE patients than in non-anergic TPE and MPE patients (p < 0.001). The percentage of CD4+CCR10+T cells in PF was higher in anergic TPE patients than in non-anergic TPE and MPE patients (p = 0.001). We reported here that CCL27/CCR10 interactions might contribute to pathophysiology in anergic TPE. PF CCL27 and CD4+CCR10+T cells may help in diagnosing TPE in patients with moderate elevation of PF ADA levels.
Asunto(s)
Adenosina Desaminasa/análisis , Quimiocina CCL27/análisis , Péptidos y Proteínas de Señalización Intercelular/análisis , Derrame Pleural/inmunología , Tuberculosis Pleural/inmunología , Adulto , Anciano , Anciano de 80 o más Años , Biomarcadores/análisis , Recuento de Linfocito CD4 , Linfocitos T CD4-Positivos/inmunología , Femenino , Citometría de Flujo , Humanos , Masculino , Persona de Mediana Edad , Derrame Pleural/diagnóstico , Derrame Pleural/microbiología , Valor Predictivo de las Pruebas , Estudios Prospectivos , Análisis por Matrices de Proteínas , Receptores CCR10/análisis , Tuberculosis Pleural/diagnóstico , Tuberculosis Pleural/microbiologíaAsunto(s)
Biomarcadores de Tumor/análisis , Antígenos CD4/análisis , Antígenos CD8/análisis , Antígenos Comunes de Leucocito/análisis , Linfoma Cutáneo de Células T/inmunología , Linfoma de Células T Periférico/inmunología , Receptores CCR10/análisis , Neoplasias Cutáneas/inmunología , Anciano , Antineoplásicos/uso terapéutico , Biomarcadores de Tumor/genética , Progresión de la Enfermedad , Resultado Fatal , Humanos , Linfoma Cutáneo de Células T/tratamiento farmacológico , Linfoma Cutáneo de Células T/genética , Linfoma Cutáneo de Células T/patología , Linfoma de Células T Periférico/tratamiento farmacológico , Linfoma de Células T Periférico/genética , Linfoma de Células T Periférico/patología , Masculino , Neoplasias Cutáneas/tratamiento farmacológico , Neoplasias Cutáneas/genética , Neoplasias Cutáneas/patología , Resultado del Tratamiento , Vorinostat/uso terapéuticoRESUMEN
Tumor cells in cutaneous T-cell lymphoma express limited numbers of chemokine receptors. We investigated the expression patterns of CXCR3, CCR3, CCR4 and CCR10 in mycosis fungoides, Sézary syndrome, lym-phomatoid papulosis and anaplastic large cell lymphoma in 121 skin biopsy samples. CXCR3 was expressed in 86% of mycosis fungoides cases but in no anaplastic large cell lymphoma cases. CCR3 was expressed in 73% of cases of CD30+ lymphoproliferative disorders such as lymphomatoid papulosis and anaplastic large cell lymphoma. Mycosis fungoides/Sézary syndrome patients with high CCR3 or CCR4 expression had a poorer survival prognosis than mycosis fungoides/Sézary syndrome patients whose tumor cells did not express these receptors. CCR10 was expressed in 50% of mycosis fungoides/Sézary syndrome cases and in 13% of cases with CD30+ lym-phoproliferative disorders. These results suggest that differential patterns of CXCR3, CCR3, CCR4 and CCR10 expression are useful for the diagnosis of cutaneous T-cell lymphoma. Moreover, expression of CCR3 or CCR4 suggests a poor prognosis in mycosis fungoides/Sézary syndrome.
Asunto(s)
Biomarcadores de Tumor/análisis , Micosis Fungoide/inmunología , Receptores CCR3/análisis , Receptores CCR4/análisis , Síndrome de Sézary/inmunología , Neoplasias Cutáneas/inmunología , Humanos , Micosis Fungoide/mortalidad , Micosis Fungoide/patología , Pronóstico , Receptores CCR10/análisis , Receptores CXCR3/análisis , Síndrome de Sézary/mortalidad , Síndrome de Sézary/patología , Neoplasias Cutáneas/mortalidad , Neoplasias Cutáneas/patología , Regulación hacia ArribaRESUMEN
Regulatory T cells (Tregs) have been suggested to play a role in the pathogenesis of atopic dermatitis (AD). However, alterations in the ability of Tregs remain to be determined. To investigate the expression of various surface receptors on CD4(+)CD25(high) regulatory T cells and to investigate their capacity for inhibiting the proliferation of CD4(+) CD25(-) effector T cells (Teffs). Peripheral blood samples were obtained from 15 patients with severe atopic dermatitis (AD) and 20 control subjects. FACs was then carried out to analyze the expression levels of FoxP3, CD152 (CTLA-4), CD39, CD73, CD223 (LAG-3), CCR4, CCR5, and CCR10 on Tregs. The proliferative responses of Teffs were assessed in the absence or presence of autologous Tregs and the TGF-ß1 and IL-10 levels in the culture supernatant and sera were detected by enzyme-linked immunosorbent assay (ELISA). The CD152, CD39, CD73, CCR4, and CCR5 expression levels on Tregs were higher in patients with severe AD than in the controls. Tregs showed an attenuated suppressive function of the proliferation of autologous Teffs in severe AD. The concentrations of IL-10 and TGF-ß in the culture supernatants of Tregs were lower in the AD group than in the control. The attenuated ability of Tregs to suppress Teff proliferation may be responsible for the autoimmune reaction of severe AD.
Asunto(s)
Dermatitis Atópica/sangre , Interleucina-10/metabolismo , Linfocitos T Reguladores/química , Linfocitos T Reguladores/inmunología , Factor de Crecimiento Transformador beta1/metabolismo , 5'-Nucleotidasa/análisis , Adulto , Antígenos CD/análisis , Apirasa/análisis , Antígeno CTLA-4/análisis , Estudios de Casos y Controles , Proliferación Celular , Células Cultivadas , Femenino , Factores de Transcripción Forkhead/análisis , Proteínas Ligadas a GPI/análisis , Humanos , Interleucina-10/sangre , Masculino , Receptores CCR10/análisis , Receptores CCR4/análisis , Receptores CCR5/análisis , Índice de Severidad de la Enfermedad , Factor de Crecimiento Transformador beta1/sangre , Proteína del Gen 3 de Activación de LinfocitosRESUMEN
The mechanisms by which CC chemokine receptor (CCR)7 ligands are selectively presented on lymphatic endothelium in the presence of inflammatory chemokines are poorly understood. The chemokine-scavenging receptor D6 is expressed on lymphatic endothelial cells (LEC) and contributes to selective presentation of CCR7 ligands by suppressing inflammatory chemokine binding to LEC surfaces. As well as preventing inappropriate inflammatory cell attachment to LECs, D6 is specifically involved in regulating the ability of LEC to discriminate between mature and immature dendritic cells (DCs). D6 overexpression reduces immature DC (iDC) adhesion to LECs, whereas D6 knockdown increases adhesion of iDCs that displace mature DCs. LEC D6 expression is regulated by growth factors, cytokines, and tumor microenvironments. In particular, interleukin-6 and interferon-γ are potent inducers, indicating a preferential role for D6 in inflamed contexts. Expression of the viral interleukin-6 homolog from Kaposi sarcoma-associated herpesvirus is also sufficient to induce significant D6 upregulation both in vitro and in vivo, and Kaposi sarcoma and primary effusion lymphoma cells demonstrate high levels of D6 expression. We therefore propose that D6, which is upregulated in both inflammatory and tumor contexts, is an essential regulator of inflammatory leukocyte interactions with LECs and is required for immature/mature DC discrimination by LECs.
Asunto(s)
Células Endoteliales/metabolismo , Receptores CCR10/genética , Receptores CCR10/fisiología , Animales , Células CHO , Comunicación Celular/genética , Comunicación Celular/inmunología , Diferenciación Celular/genética , Diferenciación Celular/inmunología , Células Cultivadas , Cricetinae , Cricetulus , Células Dendríticas/inmunología , Células Dendríticas/metabolismo , Células Dendríticas/fisiología , Células Endoteliales/inmunología , Células HEK293 , Humanos , Inflamación/genética , Inflamación/inmunología , Inflamación/metabolismo , Neoplasias/genética , Neoplasias/inmunología , Neoplasias/metabolismo , Receptores CCR10/análisis , Receptores CCR10/metabolismo , Transfección , Receptor de Quimiocina D6RESUMEN
Atypical chemokine receptors are a distinct subset of chemokine receptors able to modulate immune responses by acting as chemokine decoy/scavengers or transporters. Intracellular trafficking properties sustained by Gαi-independent signaling have emerged as a major determinant of their biological properties, which support continuous uptake, transport, and/or concentration, of the ligands. Here, we are providing methods to study both trafficking and signaling of this class of chemokine receptors focusing on the atypical chemokine receptor D6 that degrades inflammatory CC chemokines.
Asunto(s)
Citometría de Flujo/métodos , Microscopía Confocal/métodos , Receptores CCR10/análisis , Receptores CCR10/metabolismo , Animales , Quimiocinas CC/inmunología , Humanos , Immunoblotting/métodos , Proteínas de Unión al GTP Monoméricas/análisis , Proteínas de Unión al GTP Monoméricas/genética , Proteínas de Unión al GTP Monoméricas/metabolismo , Transporte de Proteínas , Receptores CCR10/inmunología , Transducción de Señal , Transfección/métodos , Receptor de Quimiocina D6RESUMEN
PURPOSE: The purpose of this study was to identify protein markers present in subjects with temporomandibular joint disorders (TMDs) and clicking compared with the levels in controls. MATERIALS AND METHODS: This was a pilot case-control study, and we report the preliminary results. Samples of joint aspirate collected from patients with TMDs and controls who had undergone surgery for a problem other than TMDs were analyzed using isobaric tags for relative and absolute quantitation (iTRAQ) and biotin-labeled-based protein arrays. The data obtained from these techniques were used to identify the proteins of interest, which were then quantitated using enzyme-linked immunosorbent assay (ELISA). The patient samples studied included joint aspirate collected clinically from the controls and patients and included samples from both the right and the left sides of each patient with a TMD. RESULTS: The 8 TMJ aspirate samples from 6 subjects included 5 aspirate samples from 4 patients and 3 from 2 controls. The greatest standardized protein concentration of endocrine gland-derived vascular endothelial growth factor/prokineticin-1 (EG-VEGF/PK1) and D6 was found in both joints of the controls compared with the levels from the joints of the patients. With 1 exception, the standardized protein concentration was significantly lower in the patients than in the controls. The lower levels of EG-VEGF/PK1 and D6 in the patients compared with the controls suggest that these cytokines might be possible biomarkers for TMDs. CONCLUSION: In the present pilot study, greater levels of EG-VEGF/PK1 and D6 were found in the controls than in the patients with TMDs. Proteomic analysis of the proteins present in the diseased joints compared with those in the controls might help to identify proteins present when pain or degeneration of the joint occurs. The proteomic information might be useful in the development of future therapies.
Asunto(s)
Biomarcadores/análisis , Proteoma/análisis , Trastornos de la Articulación Temporomandibular/diagnóstico , Activinas/análisis , Adolescente , Adulto , Anhidrasas Carbónicas/análisis , Estudios de Casos y Controles , Quimiocina CCL21/análisis , Femenino , Humanos , Factor II del Crecimiento Similar a la Insulina/análisis , Luxaciones Articulares/diagnóstico , Luxaciones Articulares/metabolismo , Metaloproteinasa 16 de la Matriz/análisis , Paracentesis , Peroxirredoxinas/análisis , Proyectos Piloto , Análisis por Matrices de Proteínas , Receptores CCR10/análisis , Líquido Sinovial/química , Disco de la Articulación Temporomandibular/patología , Trastornos de la Articulación Temporomandibular/metabolismo , Factor de Crecimiento Endotelial Vascular Derivado de Glándula Endocrina/análisis , Adulto Joven , Globinas alfa/análisis , Globinas beta/análisis , gamma-Globinas/análisis , Receptor de Quimiocina D6RESUMEN
CCL27 is one of the CC chemokines produced by epidermal keratinocytes and is suggested to be involved in the pathogenesis of inflammatory skin diseases. To clarify the contribution of CCL27 in skin inflammation, we created transgenic C57BL/6 mice that constitutively produce CCL27 in epidermal keratinocytes. These mice had high serum CCL27 levels and did not show any phenotypical change. Thus we stimulated these mice with various reagents by single and repeated application. Interestingly, only contact hypersensitivity to repeated application with fluorescein isothiocyanate was significantly enhanced in transgenic mice compared to non-transgenic mice. Under this condition, the numbers of inflammatory cells, CCR10-positive cells, CCR4-positive cells and cutaneous lymphocyte-associated antigen-positive cells were increased, and IL-4 mRNA expression was higher in the lesional skin of transgenic mice. Increased number of mast cells and higher serum IgE levels, which were similar to atopic dermatitis, were also observed. These results indicated that CCL27 modified inflammation by attracting CCR10-positive and CCR4-positive cells into the lesional skin, and may participate in the pathogenesis of Th2-shifted skin diseases such as atopic dermatitis.
Asunto(s)
Quimiocina CCL27/metabolismo , Dermatitis Alérgica por Contacto/inmunología , Células Th2/inmunología , Animales , Recuento de Células , Quimiocina CCL27/sangre , Quimiocina CCL27/genética , Quimiotaxis/efectos de los fármacos , Medios de Cultivo Condicionados/farmacología , Dermatitis Alérgica por Contacto/metabolismo , Dermatitis Alérgica por Contacto/patología , Epidermis/metabolismo , Expresión Génica/efectos de los fármacos , Inmunización/métodos , Inmunoglobulina E/sangre , Inmunoglobulina E/inmunología , Interleucina-4/genética , Queratinocitos/metabolismo , Leucocitos Mononucleares/química , Leucocitos Mononucleares/patología , Mastocitos/metabolismo , Mastocitos/patología , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Oxazolona/inmunología , Oxazolona/farmacología , Receptores CCR10/análisis , Receptores CCR4/análisis , Piel/metabolismo , Piel/patología , Células TH1/inmunología , Células TH1/metabolismo , Células Th2/metabolismoRESUMEN
Although the mucosal and the systemic immune compartments are structurally and functionally independent, they engage in cross-talk under specific conditions. To investigate this cross-talk, we vaccinated mice with tetanus toxoid together with cholera toxin with s.c. priming followed by intrarectal (IR) boosting. Interestingly, higher numbers of Ag-specific IgA and IgG Ab-secreting cells (ASCs) were detected in the lamina propria of the large intestine of mice vaccinated s.c.-IR. Ag-specific ASCs from the colon migrated to SDF-1alpha/CXCL12 and mucosae-associated epithelial chemokine/CCL28, suggesting that CXCR4(+) and/or CCR10(+) IgA ASCs found in the large intestine after s.c.-IR are of systemic origin. In the colonic patches-null mice, IgA ASCs in the large intestine were completely depleted. Furthermore, the accumulation of IgA ASCs in the colonic patches by inhibition of their migration with FTY720 revealed that colonic patches are the IgA class-switching site after s.c.-IR. Most interestingly, s.c.-IR induced numbers of Ag-specific IgA ASCs in the large intestine of TLR2(-/-), TLR4(-/-), MyD88(-/-), and TRIF(-/-) mice that were comparable with those of wild-type mice. Taken together, our results suggest the possibility that cross-talk could occur between the large intestine and the systemic immune compartments via the colonic patches without the assistance of innate immunity.