RESUMEN
In the present study we investigated the effect of bovine conglutinin on the phagocytic activity of leukocytes. We measured both the chemotactic activity of conglutinin and its effect on the internalization of zymosan particles and E. coli by granulocytes. We also assessed the binding of conglutinin to various microorganisms isolated from clinical cases in cattle. We showed that conglutinin binds strongly to the surface of yeast cells and to mannan-rich zymosan particles, while weak binding was observed in the case of the bacterial strains tested, including those whose O antigen is composed of mannan. Conglutinin (1-10 microg/ml) neither acts as a chemotactic factor for peripheral blood leukocytes nor affects ingestion of E. coli by granulocytes. However, as flow cytometry based assay showed, conglutinin (0.1-1 microg/ml) increased ingestion of zymosan expressed as mean fluorescence intensity (MFI) of positive cells.
Asunto(s)
Colectinas/farmacología , Granulocitos/efectos de los fármacos , Fagocitosis/efectos de los fármacos , Seroglobulinas/farmacología , Animales , Bacterias , Candida albicans , Bovinos , Quimiotaxis/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Granulocitos/fisiología , Humanos , Fagocitosis/fisiología , Unión Proteica , Zimosan/químicaRESUMEN
The aim of our study was to estimate both B7-H1 and B7-H4 molecules on immature myeloid and lymphoid dendritic cells in umbilical cord blood of healthy neonates in comparison with peripheral blood of healthy adults. Thirty nine healthy full-term neonates from physiological single pregnancies and 27 healthy adults were included in the study. The expression of B7-H1 and B7-H4 was revealed using the immunofluorescence method. Statistical analysis was performed using a non-parametric test (Mann-Whitney U-Test). The percentages of BDCA-1+ dendritic cells with B7-H1 and B7-H4 expressions were significantly higher in peripheral blood of healthy adults (p<0.00003). It was either observed that the percentage of BDCA-2+ dendritic cells with the expression of B7-H4 molecules was significantly higher in peripheral blood of healthy adults in comparison with umbilical cord blood (p<0.02). Decreased percentages of dendritic cells and co-stimulatory molecules indicate that neonates have immature immune system. Depletion of co-stimulatory B7-H1 and B7-H4 molecules enable appropriate development of immune response.
Asunto(s)
Antígenos CD/metabolismo , Antígeno B7-1/metabolismo , Células Dendríticas/metabolismo , Sangre Fetal/metabolismo , Linfocitos/metabolismo , Células Mieloides/metabolismo , Adulto , Antígeno B7-H1 , Células Dendríticas/inmunología , Citometría de Flujo , Humanos , Recién Nacido , Lectinas Tipo C/metabolismo , Linfocitos/inmunología , Glicoproteínas de Membrana/metabolismo , Células Mieloides/inmunología , Receptores Inmunológicos/metabolismo , Inhibidor 1 de la Activación de Células T con Dominio V-SetRESUMEN
Disturbed immunoregulation and an inappropriate immune response to gut microflora is assumed to be involved in the pathogenesis of inflammatory bowel disease (IBD). Physiologically dendritic cells (DCs) as the professional antigen presenting cells play a crucial role in the control of intestinal inflammation and immune tolerance. In order to evaluate their role in the IBD we analyzed the phenotypic and functional properties of monocyte-derived DCs (MoDCs) generated from UC and CD patients following stimulation with TNF-α, lipopolisaccharide E. coli or hydrocortisone. Thirty seven patients with moderate to severe inflammation (19 UC, 18 CD) were recruited to the study. Monocyte-derived dendritic cell immunophenotypes and their endocytic ability were analysed by flow cytometry and confocal microscopy, IL-6, IL-10, IL-12 and IL-23 secretion were investigated by ELISA. Both unstimulated and stimulated MoDCs generated from IBD patients had more mature phenotype and secreted elevated concentrations of proinflammatory cytokines as compared to a control group. The addition of LPS E. coli to culture media was associated with enhanced dendritic cell activation and maturation as compared to DCs stimulated only with TNF-α. This may suggest altered dendritic cell interactions with intestinal microflora in inflammatory bowel disease. Hydrocortisone decreases the numbers of mature dendritic cells and the proinflammatory cytokine concentrations in all cell culture types that may explain the efficacy of steroid therapy in inflammatory bowel disease.
Asunto(s)
Colitis Ulcerosa/inmunología , Enfermedad de Crohn/inmunología , Células Dendríticas/inmunología , Monocitos/inmunología , Adulto , Antígenos/inmunología , Antígenos/metabolismo , Colitis Ulcerosa/metabolismo , Colitis Ulcerosa/patología , Enfermedad de Crohn/metabolismo , Enfermedad de Crohn/patología , Citocinas/inmunología , Citocinas/metabolismo , Células Dendríticas/metabolismo , Células Dendríticas/patología , Escherichia coli/inmunología , Femenino , Humanos , Hidrocortisona/inmunología , Hidrocortisona/metabolismo , Inmunofenotipificación/métodos , Lipopolisacáridos/inmunología , Lipopolisacáridos/metabolismo , Masculino , Persona de Mediana Edad , Monocitos/metabolismo , Monocitos/patología , Factor de Necrosis Tumoral alfa/inmunología , Factor de Necrosis Tumoral alfa/metabolismo , Adulto JovenRESUMEN
PPAR-γ plays a role in the development of immune response, particularly in inflammation. The inflammatory reaction may be stimulated or suppressed by the presence of PPAR ligands. Some researchers suggest positive influence of the PPAR-γ agonist on suppression of the intestinal inflammatory process, yet there has not been much evidence showing that the antagonist of PPAR-γ can affect the inflammatory process. The aim of the present study was to define the mechanism by which PPAR-γ ligands affect the course of experimentally induced colitis in rats. Colitis was induced in rats by rectal administration of TNBS (trinitrobenzene sulfonate). Rosiglitazone was administrated to animals at the dose of 8 mg/kg four times via an intra-gastric probe. Biphenol-A-diglicydyl ether (BADGE) was administrated intraperitoneally at the dose of 120 mg/kg, three times every second day. One group of animals received rosiglitazone together with BADGE before the induction of inflammation. Histological and ELISA examinations of large intestine samples were performed. Levels of IL-1ß, IL-6, TNF-α cytokines were determined in serum and homogenates. Rats exposed to rosiglitazone had higher body weight yet lower large intestine weight. Histological findings showed less ulceration, lower expression of crypts' loss and smaller oedema. Animals, which did not receive rosiglitazone, and those receiving it together with BADGE, developed more severe inflammatory changes. Rosiglitazone decreased the expression of inflammatory cytokines, such as IL-6 and TNF-α, both in serum and in intestinal homogenates. BADGE used with TNBS did not increase the expression of inflammatory cytokines; however, applied together with rosiglitazone, it caused inflammation similar to that observed among rats with experimentally induced colitis. Rosiglitazone reduces inflammation by decreasing the expression of IL-6 and TNF-α. BADGE administered with rosiglitazone blocks the activity of PPAR-γ and abolishes the protective effects of PPAR-γ agonist.
Asunto(s)
Colitis/tratamiento farmacológico , Compuestos Epoxi/farmacología , PPAR gamma/agonistas , PPAR gamma/antagonistas & inhibidores , Tiazolidinedionas/farmacología , Animales , Compuestos de Bencidrilo , Colitis/inmunología , Colitis/metabolismo , Colitis/patología , Citocinas/biosíntesis , Citocinas/metabolismo , Intestino Grueso/metabolismo , Intestino Grueso/patología , PPAR gamma/inmunología , PPAR gamma/metabolismo , Ratas , Ratas Wistar , RosiglitazonaRESUMEN
Recently, we described that vaccination with allogeneic dendritic cells (DCs) pulsed with tumor cell lysate generated specific CD8+ T cell response in patients with B-cell chronic lymphocytic leukemia (B-CLL). In the present study, the potential of autologous DCs pulsed ex vivo with tumor cell lysates to stimulate antitumor immunity in patients with B-CLL in early stages was evaluated. Twelve patients at clinical stage 0-2 as per Rai were vaccinated intradermally up to eight times with a mean number of 7.4 x 10(6) DCs pulsed with B-CLL cell lysate. We observed a decrease of peripheral blood leukocytes and CD19+/CD5+ leukemic cells in five patients, three patients showed a stable disease and four patients progressed despite DC vaccination. A significant increase of specific cytotoxic CD8+ T lymphocytes against the leukemia-associated antigens RHAMM or fibromodulin was detected in four patients after DC vaccination. In patients with a clinical response, an increase of interleukin 12 (IL-12) serum levels and a decrease of the frequency of CD4+CD25(+)FOXP3+ T regulatory cells were observed. Taken together, the study demonstrated that vaccination with autologous DC in CLL patients is feasible and safe. Immunological and to some extend hematological responses could be noted, justifying further investigation on this immunotherapeutical approach.