RESUMEN
Neutrophils play important roles in many inflammatory diseases. The migration of neutrophils to the inflammatory site is tightly regulated by specific chemokines, of which interleukin-8 (IL-8) and leukotriene B4 (LTB4 ) constitute key mediators by binding to the surface receptors CXCR1/2 and BLT1, respectively. Oligonucleotides (ODN) containing CpG motifs mediate potent immunomodulatory effects through binding to Toll-like receptor 9. So far, knowledge on how ODN can affect neutrophil migration during inflammation is lacking. This study demonstrates that several novel CpG ODN significantly down-regulate the surface expression of CXCR1/2 and BLT1. In addition, the ODN significantly blocked IL-8-induced and LTB4 -induced neutrophil migration in vitro, as well as leucocyte migration in vivo demonstrated in mice by intravital microscopy and in a model of airway inflammation. The down-regulation of CXCR1 is rapid, occurring 15 min after ODN stimulation, and can be mediated through an endosomally independent mechanism. Inhibition of the IL-8 and LTB4 pathways may provide new opportunities of therapeutic intervention using ODN to reduce neutrophil infiltration during inflammation.
Asunto(s)
Factores Inmunológicos/farmacología , Inmunomodulación , Infiltración Neutrófila/efectos de los fármacos , Neutrófilos/efectos de los fármacos , Oligonucleótidos/farmacología , Receptores de Interleucina-8A/biosíntesis , Receptores de Interleucina-8B/biosíntesis , Receptores de Leucotrieno B4/inmunología , Animales , Quimiotaxis de Leucocito/efectos de los fármacos , Quimiotaxis de Leucocito/inmunología , Islas de CpG/genética , Regulación hacia Abajo/inmunología , Femenino , Humanos , Inflamación/tratamiento farmacológico , Inflamación/inmunología , Interleucina-8/antagonistas & inhibidores , Interleucina-8/biosíntesis , Interleucina-8/inmunología , Antígeno de Macrófago-1/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Infiltración Neutrófila/inmunología , Neutrófilos/inmunología , Ovalbúmina , Receptores de Leucotrieno B4/antagonistas & inhibidores , Receptores de Leucotrieno B4/biosíntesis , Receptor Toll-Like 9/inmunología , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
BACKGROUND: Glucocorticoids (GCS) remain one of the mainstay treatments in the management of ulcerative colitis (UC) but up to a third of patients will ultimately fail to respond and progress to a more severe and difficult to manage disease state. Previous clinical studies suggest that the Toll-Like Receptor 9 (TLR9) agonist DIMS0150 not only induces production of key anti-inflammatory cytokines as IL-10 but interestingly also enhances steroid sensitivity in steroid refractory UC patients. We investigated, in the context of a clinical study, whether a pre-selection of steroid response genes could identify steroid refractory UC subjects most likely to respond to DIMS0150 treatment. METHODS: In a non-interventional pilot study, blood from steroid refractory UC patients and healthy volunteers was taken and thirty-four previously described steroid response genes were analysed by real time PCR analysis. To establish clinical utility of the identified biomarkers, a placebo controlled, randomized, double blinded study in active steroid dependent and steroid resistant UC patients on concomitant steroid therapies was used (EudraCT number: 2006-001846-15). RESULTS: We identified three potential biomarkers CD163, TSP-1 and IL-1RII whose response to steroids was significantly enhanced when DIMS0150 was applied. Thirty-four subjects were randomized to receive a single rectal administration of placebo or 30 mg of DIMS0150. Blood derived PBMCs were obtained prior to dosing and assayed for evidence of a steroid enhancing effect following steroid incubation in the presence of DIMS0150. Comparison to established steroid sensitivity marker IL-6 confirmed that clinical responders are steroid refractory UC patients. Upon study completion and un-blinding, the biomarker assay correctly predicted a clinical response in over 90% of the patients. CONCLUSION: Using specific steroid response biomarkers, GCS refractory UC patients most likely to benefit from DIMS0150 treatment could be identified and illustrates the usefulness of a personalized treatment approach.
Asunto(s)
Colitis Ulcerosa/tratamiento farmacológico , ADN/uso terapéutico , Glucocorticoides/uso terapéutico , Factores Inmunológicos/uso terapéutico , Receptor Toll-Like 9/agonistas , Administración Rectal , Adulto , Anciano , Antígenos CD/genética , Antígenos de Diferenciación Mielomonocítica/genética , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/genética , Biomarcadores , Estudios de Casos y Controles , Colitis Ulcerosa/genética , Método Doble Ciego , Femenino , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Pronóstico , Receptores de Superficie Celular/genética , Receptores Tipo II de Interleucina-1/genética , Resultado del Tratamiento , Adulto JovenRESUMEN
BACKGROUND & AIMS: Inflammatory bowel diseases (IBDs) and the irritable bowel syndrome (IBS) are heterogeneous disorders of the gastrointestinal tract and can profoundly affect the quality of life. Because many of the symptoms of IBD are similar to those of IBS, the former may be misdiagnosed. In addition, the 2 major forms of IBD, ulcerative colitis (UC) and Crohn's disease (CD), have overlapping nonspecific, pathologic features leading to difficulties in assessing colonic inflammation and hence the term IBD unclassified has been proposed. The aim of this study was to identify and assess the utility of a certain set of marker genes that could help to distinguish IBS from IBD, and further to discriminate between UC and CD. METHODS: Subtractive suppression hybridization was used to identify IBD-specific genes in colonic mucosal biopsy specimens. In quantitative polymerase chain reaction experiments, the differential expressions of identified genes then were analyzed using a classification algorithm and the possible clinical value of these marker genes was evaluated in a total of 301 patients in 3 stepwise studies. RESULTS: Seven marker genes were identified as differentially expressed in IBD, making it possible to discriminate between patients suffering from UC, CD, or IBS with area under the receiver-operating characteristic curves ranging from 0.915 to 0.999 (P < .0001) using the clinical diagnosis as gold standard. CONCLUSIONS: Expression profiling of relevant marker genes in colonic biopsy specimens from patients with IBD/IBS-like symptoms may enable swift and reliable determination of diagnosis, ultimately improving disease management.
Asunto(s)
Colitis Ulcerosa/genética , Enfermedad de Crohn/genética , Perfilación de la Expresión Génica , Marcadores Genéticos , Pruebas Genéticas , Síndrome del Colon Irritable/genética , Adulto , Anciano , Anciano de 80 o más Años , Algoritmos , Colitis Ulcerosa/diagnóstico , Colitis Ulcerosa/patología , Colon/química , Colon/patología , Enfermedad de Crohn/diagnóstico , Enfermedad de Crohn/patología , ADN Complementario/análisis , Diagnóstico Diferencial , Femenino , Perfilación de la Expresión Génica/métodos , Predisposición Genética a la Enfermedad , Genotipo , Humanos , Mucosa Intestinal/química , Mucosa Intestinal/patología , Síndrome del Colon Irritable/diagnóstico , Síndrome del Colon Irritable/patología , Masculino , Persona de Mediana Edad , Análisis de Secuencia por Matrices de Oligonucleótidos , Fenotipo , Proyectos Piloto , Valor Predictivo de las Pruebas , Estudios Prospectivos , ARN/análisis , Curva ROC , Reproducibilidad de los Resultados , Estudios Retrospectivos , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
BACKGROUND: Patients with chronic active ulcerative colitis (UC) are regarded as treatment failures and represent an area of high unmet medical need, as normally the only remaining option is colectomy. METHODS: We treated a total of eight chronic active severe UC outpatients with the immunomodulatory agent DIMS0150 as an add-on to current therapies. Seven patients received a single topical dose of 30 mg and one special case subject received three doses with 4 weeks between dosing occasions. All patients were classed as treatment failures and were elected for colectomy. Efficacy evaluation was determined in terms of colitis activity index, endoscopic improvement, and histologic disease activity assessed primarily at week 12 with a follow-up period of over 2 years. Glucocorticoid sensitivity was assayed by in vitro measurement of interleukin 6. RESULTS: All patients demonstrated a pronounced and rapid reduction in their colitis activity index within 1 week following a single intracolonic administration via colonoscope of the agent DIMS0150. Further improvements were evident at week 4, resulting in a clinical response rate for the single-dose treatment of 71%, with 43% in clinical remission. By week 12 the clinical response and remission rates had reached 82% and 71%, respectively. A follow-up period of over 2 years posttreatment indicated that all but one of the treated patients had avoided the need for colectomy, with the longest patient being in symptom-free remission for over 27 months. Treatment with DIMS0150 restored glucocorticoid sensitivity. CONCLUSIONS: DIMS0150 may have the potential to be an effective agent to treat chronic active UC patients with the prospect to avoid colectomy on a long-term basis and is currently the subject of a clinical phase III study (EudraCT number: 2011-003130-14).
Asunto(s)
Colitis Ulcerosa/tratamiento farmacológico , ADN/uso terapéutico , Glucocorticoides/uso terapéutico , Factores Inmunológicos/uso terapéutico , Prednisolona/uso terapéutico , Administración Tópica , Adulto , Anciano , Biomarcadores/sangre , Colitis Ulcerosa/metabolismo , Colon/metabolismo , Colonoscopía , Esquema de Medicación , Quimioterapia Combinada , Femenino , Estudios de Seguimiento , Humanos , Interleucina-6/sangre , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Receptor Toll-Like 9/metabolismo , Resultado del TratamientoRESUMEN
BACKGROUND AND AIMS: Inflammatory bowel disease (IBD) is a complex inflammatory disease of the gastrointestinal tract with unknown cause that lacks molecular markers for diagnosis. Crohn's disease (CD) and ulcerative colitis (UC) are the two major forms of IBD. The aim of this study was to investigate gene expression patterns in UC and characterize newly identified marker genes potentially linked to disease pathogenesis of UC. MATERIALS AND METHODS: Biopsies were taken from eight UC patients, from inflamed and non-inflamed parts of the colon. Gene expression was investigated by subtractive suppression hybridization (SSH), and further study of a selected gene was performed by Northern blot, immunohistochemistry, immunocytochemistry, and in vitro monocyte differentiation. RESULTS: Three hundred thirty-one differentially expressed genes were found and classified into functional groups. In this paper, we report one gene with unknown function to be differentially expressed in UC but not Crohn's disease by real-time reverse transcriptase polymerase chain reaction. Due to its predicted protein architecture, we call this gene Wiskott-Aldrich syndrome protein and FKBP-like (WAFL). Initial pilot experiments suggest WAFL to participate in innate immune functions. CONCLUSION: The SSH result supports the current view of UC to be a chronic inflammatory disorder with aberrant expression of epithelial barrier proteins, cell fate-related factors, and disturbed metabolism. The new gene, WAFL, reported in this study, appears to be conditionally regulated in myeloid cells. This indicates that WAFL may be connected to innate immune-host responses. As such, it represents an interesting, hitherto unknown player in IBD where there is a need for further elucidation on the molecular and cellular level.