RESUMEN
BACKGROUND: This study investigated whether a dot immunoassay (DIA) can provide simultaneous detection of anti-tissue transglutaminase (tTG), anti-deamidated gliadin (DG) and total IgA antibodies, as required in the work-up of celiac disease (CD) patients. METHODS: Celiac disease patients (n=111) consecutively diagnosed from 2001 to 2011 at the Children's Hospital and Institute of Immunology (Technical University Dresden) were tested for anti-tTG, anti-DG and total IgA by enzyme-linked immunosorbent assay (ELISA) and DIA retrospectively. Blood donors (n=45) and non-CD individuals with low IgA serum levels (n=8) were included as controls. Antibodies to endomysial antigens (EmA) were assessed by indirect immunofluorescence (IIF). RESULTS: Four (3.6%) of 111 CD patients demonstrated an IgA deficiency with total IgA below 50 mg/L by ELISA. Total IgA of the 107 IgA-non-deficient CD patients varied from 70 to 6000 mg/L. All four IgA-deficient CD patients were detected by a reduced reaction control of DIA and demonstrated positive anti-tTG or anti-DG IgG by DIA or ELISA. Detection of anti-tTG and anti-DG by DIA and ELISA showed a very good agreement (IgA: κ=0.972, 0.856, respectively; IgG: 0.921, 0.895, respectively). CONCLUSIONS: Immunodot assay is a reliable and easy-to-use technique for the detection of IgA-deficient CD patients. Simultaneous assessment of anti-tTG and anti-DG IgA antibodies, and IgA deficiency by DIA can improve the efficacy of CD serology.
Asunto(s)
Enfermedad Celíaca/diagnóstico , Deficiencia de IgA/diagnóstico , Inmunoensayo , Adolescente , Adulto , Anciano , Enfermedad Celíaca/inmunología , Niño , Preescolar , Femenino , Humanos , Deficiencia de IgA/inmunología , Lactante , Masculino , Persona de Mediana EdadRESUMEN
Anti-double-stranded (ds)DNA antibodies are serological markers of systemic lupus erythematosus (SLE). Of all anti-dsDNA antibody detection methods, the Crithidia luciliae immunofluorescence test (CLIFT) is thought to have the highest specificity for SLE. However, the clinical application is hampered by the low diagnostic sensitivity. A CLIFT with modified assay buffer (mCLIFT) was developed and compared with conventional CLIFT, using sera from 110 patients with SLE, 89 anti-dsDNA ELISA-positive patients with other diseases (non-SLE group A), 157 non-SLE patients with undetectable anti-dsDNA antibodies by ELISA (non-SLE group B), 77 disease controls (non-SLE group C), and 50 healthy blood donors. Out of the 110 anti-dsDNA antibody ELISA-positive SLE patients, 84 (76.4%) demonstrated a positive kinetoplast staining, using the mCLIFT, compared to only 42.3%, using the conventional CLIFT. The diagnostic specificity of mCLIFT was 100% with healthy blood donors and 98.1% with the non-SLE group C (anti-nuclear antibodies negative; no signs or symptoms of an autoimmune disease) included. In the non-SLE groups A and B with various other autoimmune diseases or symptoms of a possible autoimmune disease, positive mCLIFT results were obtained in 33.7% and 3.2%, respectively. In conclusion, by modification of the assay buffer, a significant increase in sensitivity of the CLIFT could be observed while retaining the high specificity for SLE. Further investigation is required to check whether the CLIFT-positive non-SLE patients develop SLE and whether anti-dsDNA antibodies detected by the mCLIFT represent a pathogenetic and diagnostic subgroup of autoantibodies that may improve the early diagnosis of SLE or SLE-overlap syndromes.
Asunto(s)
Anticuerpos Antinucleares/sangre , Crithidia/inmunología , Técnica del Anticuerpo Fluorescente/métodos , Lupus Eritematoso Sistémico/diagnóstico , Animales , Antígenos de Protozoos/inmunología , Ensayo de Inmunoadsorción Enzimática , Humanos , Microscopía Fluorescente , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
INTRODUCTION: Complex perioperative immunodysfunction occurs in patients with renal cell carcinoma undergoing surgery. Here, we report on the effect of preoperative treatment with interferon-alpha2a (IFN-alpha2a). MATERIALS AND METHODS: 30 patients with a renal tumour received preoperative IFN-alpha2a for 6 days beginning 1 week before nephrectomy, 30 did not. Parameters of cellular and humoral immunity were measured in venous blood at various intervals using flow cytometry and ELISA. Endpoints included effects on immune parameters, toxicity, and survival. RESULTS: Toxicity was grade 1 in 52%, 2 in 30%, and 3 in 4%. During IFN-alpha2a administration, leukocytes, monocytes, granulocytes, B-cell marker CD19, activation markers, CD4+CD25+ regulatory T-cells, and vascular endothelial growth factor (VEGF) dropped significantly, but no difference was observed in T-cell and natural killer (NK)-cell markers, and IL-10. Postoperatively, T-cell and activation markers decreased in both groups, but CD4, CD28, IL-6, IL-10, and HLA-DR alterations were significantly less accentuated in patients who had been treated with IFN-alpha2a. After a median follow-up of 23 months, survival did not differ between the groups (p = 0.54). CONCLUSIONS: Perioperative immunodysfunction can be modulated by preoperative administration of IFN- alpha2a. IFN-alpha2a decreased the level of VEGF and CD4+CD25+ regulatory T-cells implicating a potential combination with tyrosine kinase inhibitors and vaccines.
Asunto(s)
Antineoplásicos/uso terapéutico , Carcinoma de Células Renales/tratamiento farmacológico , Carcinoma de Células Renales/cirugía , Síndromes de Inmunodeficiencia/tratamiento farmacológico , Interferón-alfa/uso terapéutico , Neoplasias Renales/tratamiento farmacológico , Neoplasias Renales/cirugía , Terapia Neoadyuvante , Adulto , Anciano , Anciano de 80 o más Años , Formación de Anticuerpos/efectos de los fármacos , Antineoplásicos/toxicidad , Antígenos CD4/sangre , Carcinoma de Células Renales/inmunología , Carcinoma de Células Renales/patología , Relación Dosis-Respuesta a Droga , Esquema de Medicación , Femenino , Humanos , Inmunidad Celular/efectos de los fármacos , Síndromes de Inmunodeficiencia/inmunología , Interferón alfa-2 , Interferón-alfa/toxicidad , Subunidad alfa del Receptor de Interleucina-2/sangre , Neoplasias Renales/inmunología , Neoplasias Renales/patología , Masculino , Persona de Mediana Edad , Estadificación de Neoplasias , Complicaciones Posoperatorias/inmunología , Proteínas Recombinantes , Linfocitos T Reguladores/efectos de los fármacos , Factor A de Crecimiento Endotelial Vascular/sangreRESUMEN
Inhibitors of alanyl-aminopeptidase e.g. phebestin increase the expression of transforming growth factor (TGF)-beta1 in mononuclear cells. We investigated whether phebestin also produced this effect in CD4+CD25+ T-cells and whether phebestin-treated CD4+CD25+ T-cells were capable of ameliorating acute colitis in mice. The suppressive activity of mouse CD4+CD25+ T-cells was assessed in vitro by co-culture with splenocytes. mRNA expression associated with the suppressive phenotype was determined in vitro and in vivo. The in vivo role of phebestin-exposed CD4+CD25+ T-cells was studied in sodium dextran sulfate-induced acute colitis in mice. The proliferation of activated effector T-cells or splenocytes in vitro was inversely correlated with the number of CD4+CD25+ T-cells. Phebestin pre-treatment substantially enhanced the suppressive activity of these cells and increased expression levels of TGF-beta1 and FoxP3. Furthermore, transfer of CD4+CD25+ T-cells exposed to phebestin for a short time ex vivo significantly reduced the mouse colitis disease activity index. We conclude that aminopeptidase inhibitors support the suppressive activity as well as TGF-beta1 and FoxP3 expression of natural regulatory T-cells.
Asunto(s)
Antígenos CD13/antagonistas & inhibidores , Antígenos CD4/metabolismo , Colitis/enzimología , Factores de Transcripción Forkhead/genética , Subunidad alfa del Receptor de Interleucina-2/metabolismo , Linfocitos T Reguladores/enzimología , Factor de Crecimiento Transformador beta1/genética , Animales , Colitis/patología , Femenino , Factores de Transcripción Forkhead/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Tolerancia Inmunológica/efectos de los fármacos , Activación de Linfocitos/efectos de los fármacos , Ratones , Ratones Endogámicos BALB C , Oligopéptidos/farmacología , Fenotipo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Linfocitos T Reguladores/citología , Linfocitos T Reguladores/efectos de los fármacos , Factor de Crecimiento Transformador beta1/metabolismoRESUMEN
Recent studies suggest the importance of prothrombotic and proinflammatory cascades in vascular thrombus formation. However, the impact of platelet CD40 and CD40 ligand (CD40L) expression and its relation to inflammatory markers in atrial clot formation have not yet been determined. Therefore, we studied a total of 40 patients. A total of 20 patients with persistent atrial fibrillation (AF) and 20 matched patients with sinus rhythm (SR) were included to quantify platelet surface expression of CD40/CD40L, serum levels of intercellular adhesion molecule-1 (ICAM), vascular adhesion molecule-1 (VCAM), high-sensitivity C-reactive protein (hsCRP), and monocyte chemoattractant protein-1 (MCP-1). Using fluorescence-activated cell sorting analysis, baseline CD40 expression (antibody binding capacity [ABC]) was increased during AF (AF: 7776 +/- 8.46 ABC vs. SR: 7753 +/- 7.32 ABC; P < 0.05), whereas CD40L expression was not different. In contrast to the effect of adenosine diphosphate, ex vivo stimulation with thrombin receptor activating peptide (TRAP) increased CD40 and CD40L expression in both groups. MCP-1, hsCRP, ICAM, and VCAM levels were significantly increased during AF, reaching highest levels in patients with atrial thrombi. Importantly, VCAM and MCP-1 were independent predictors for atrial thrombi (P < 0.05) using multivariate analysis. In contrast to declining levels of hsCRP, levels of ICAM, VCAM, MCP-1, and platelet CD40 expression remained elevated 5 weeks after successful electrical direct current cardioversion (CV). In conclusion, prothrombogenic markers are substantially elevated in patients with AF, reaching highest levels in patients with AF and atrial thrombi. Interestingly, amounts of adhesion molecules and platelet CD40 levels remain elevated even 5 weeks after successful CV, which may imply a persistently increased risk for atrial thrombus formation. In addition to hsCRP, MCP-1 and VCAM may serve as new biomarkers, which may help to identify patients with an increased risk for thromboembolic events.
Asunto(s)
Fibrilación Atrial/metabolismo , Plaquetas/metabolismo , Antígenos CD40/biosíntesis , Ligando de CD40/biosíntesis , Moléculas de Adhesión Celular/biosíntesis , Mediadores de Inflamación/sangre , Adenosina Difosfato/farmacología , Fibrilación Atrial/complicaciones , Biomarcadores/metabolismo , Femenino , Estudios de Seguimiento , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Masculino , Persona de Mediana Edad , Factores de Riesgo , Trombosis/etiología , Trombosis/metabolismo , Factores de TiempoAsunto(s)
Enfermedades Autoinmunes/terapia , Antígenos CD13/metabolismo , Colitis/terapia , Dipeptidil Peptidasa 4/metabolismo , Inflamación/terapia , Animales , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/fisiología , Colitis/inducido químicamente , Colitis/inmunología , Colitis/patología , Modelos Animales de Enfermedad , Femenino , Humanos , Activación de Linfocitos , Ratones , Ratones Endogámicos BALB C , Inhibidores de Proteasas/metabolismo , Receptores de Interleucina-2/inmunologíaRESUMEN
The human CD30-positive anaplastic large (T-) cell lymphoma cell line, KARPAS-299 (DSM ACC31), was established from blast cells in the peripheral blood from a case of non-Hodgkin lymphoma in 1988. We describe the mRNA and surface expression in KARPAS-299 cells of a panel of markers highly restricted to human natural regulatory T-cells and associated with their suppressive activity, including FOXP3, CD25, IL-10, TGF-beta1, CD62L, and Lag-3. Results obtained from co-culturing human peripheral blood leukocytes with KARPAS-299 cells assigned a suppressive phenotype to the latter ones. In conclusion, KARPAS-299 cells show characteristics typical of natural regulatory T-cells and, thus, represent a valuable model for studying regulatory T-cell function, which may also facilitate drug development aimed at the modulation of regulatory T-cell activity for the pharmacological therapy of, for example, autoimmune diseases.
Asunto(s)
Linfocitos T Reguladores/metabolismo , Línea Celular Tumoral , Técnicas de Cocultivo , Factores de Transcripción Forkhead/metabolismo , Humanos , Fenotipo , ARN Mensajero/genética , Factor de Crecimiento Transformador beta/metabolismo , Factor de Crecimiento Transformador beta1RESUMEN
STUDY OBJECTIVE: The aim of this study was to investigate the influence of 2 established anesthetic techniques: total intravenous anesthesia and balanced inhalation anesthesia (BAL) on the perioperative-induced changes of peripheral blood mononuclear cells (PBMCs), changes in lymphocyte subsets, and the balance of proinflammatory and anti-inflammatory cytokines. DESIGN: This is a prospective, randomized, clinical comparison study. SETTINGS: This study was set at a university hospital. PATIENTS: This study involved 50 patients with American Society of Anesthesiologists physical status I who were scheduled for elective minimal invasive partial diskectomy. INTERVENTIONS: There was no intervention involved in this study. MEASUREMENTS: Changes in differential counts, lymphocyte subsets, and proliferation rates were determined before surgery and in the early postoperative period. Plasma concentrations of proinflammatory cytokines (IL-2, IL-6, IL-12, interferon gamma) and anti-inflammatory cytokines (IL-10, IL-1RA, transforming growth factor beta), and plasma concentrations of cortisol, epinephrine, and norepinephrine were measured before, during, and after surgery. MAIN RESULTS: Absolute number of CD3+, CD4+, and CD8+, and expression of HLA-DR and activation marker CD25+, CD26+, and CD69+ decreased more in response to surgery after BAL. Changes in distribution of T-lymphocyte cells seem to be in part related to severe postoperative pain. Plasma concentration of IL-6 significantly increased during and after surgery with BAL without relation to pain. CONCLUSION: Anesthetic management may have varying influences on the postoperative immune response. Surgery-induced inflammatory response and alteration in cell-mediated immunity seem to be more pronounced after BAL. These effects were attributed to the enhanced stress response after BAL.
Asunto(s)
Anestesia por Inhalación , Anestesia Intravenosa , Citocinas/sangre , Inmunidad Celular/fisiología , Adulto , Proliferación Celular , Discectomía , Método Doble Ciego , Epinefrina/sangre , Femenino , Humanos , Hidrocortisona/sangre , Recuento de Leucocitos , Activación de Linfocitos/fisiología , Recuento de Linfocitos , Subgrupos Linfocitarios/inmunología , Masculino , Persona de Mediana Edad , Mitógenos/farmacología , Monocitos/inmunología , Norepinefrina/sangre , Dolor Postoperatorio/inmunologíaRESUMEN
Inhibitors of the enzymatic activity of alanyl-aminopeptidases severely affect growth and typical functions of human peripheral T cells both in vitro and in vivo. The most prominent changes observed include the activation of cellular signal transduction pathways such as MAP kinases Erk1/2 or the Wnt-pathway, a decrease of production and release of "pro-inflammatory" cytokines (IL-2, IL-12) and, most importantly, an induction of expression and release of the immunosuppressive cytokine, TGF-beta1. Similar effects on T cell proliferation and function have been observed in response to inhibition of DPIV, which is strongly suggestive of a functional synergism of APN and DPIV. In support of this hypothesis evidence is provided showing that the simultaneous application of inhibitors of DPIV and APN further enhances the anti-inflammatory and immunosuppressive effects provoked by the inhibition of APN or DPIV alone. Therefore, the simultaneous inhibition of these enzymes represents a promising strategy for the pharmacological therapy of T cell mediated diseases such as autoimmune disease, inflammation, allergy, and allograft rejection.