Background: The autoimmune subclass of chronic idiopathic urticaria (CU) has been characterized by the occurrence of biologically relevant IgG antibodies against the IgE molecule or the αchain of the high-affinity Fc...Background: The autoimmune subclass of chronic idiopathic urticaria (CU) has been characterized by the occurrence of biologically relevant IgG antibodies against the IgE molecule or the αchain of the high-affinity Fc receptor (FcRIα) on basophils and mast cells. These antibodies are usually detected by autologous serum skin testing and confirmed by histamine release studies, immunoblotting, or enzyme-linked immunosorbent assay, but not always. Objectives: To detect autoantibodies to the FcRIαin sera of CU patients by a modified serum-induced basophil activation testmeasured by flowcytometry (FCM) and to evaluate the relationship between the in vitro functional test, the autologous serum skin test (ASST), and the serum levels of IgE, eosinophil cationic protein (ECP) and antithyroid antibodies. Methods: Sera of 30 patients with CU and 26 patients with systemic autoimmune diseases (systemic lupus erythematosus, dermatomyositis) were tested for CD63 activation marker expression on basophils by FCM. Leucocytes from two highly sensitized atopic donors (DA1. DA2) and one non-atopic donor (DNA) were incubated with patients’sera and double-labelled with anti-IgE and anti-CD63 antibodies. Subsequently, the percentage of CD63-expressing basophils was determined by using FCM. In all CU patients an ASST was carried out and the serum IgE, and ECP levels and antithyroid antibodies were evaluated. Results: Twelve patients had a positive ASST and 14 patients a positive CD63 expression assay. There was a strong correlation between the ASST and CD63 assay. Sera from patients with systemic autoimmune diseases did not raise positive CD63 expression on basophils. There was a moderate negative correlation between the occurrence of atopic serum markers (IgE, ECP) and the ability of sera to induce CD63 expression on basophil cells of DA2 (P < 0.05). The female sex was preponderant and antithyroid antibodies were more frequent. Conclusions: Our new technical observation demonstrates that basophils of highly sensitized atopic donors can be successfully used without priming with IL-3 for the in-vitro flow cytofluorimetric diagnosis of CU. With this investigation the characterization of the autoimmune origin of CU is based on an objective in vitro technique.展开更多
Background: Antibodies directed to the α subunit of the high affinity IgE receptor and the IgE molecule are proposed to be of pathogenetic relevance in a group of patients with chronic urticaria (CU). The diagnosis o...Background: Antibodies directed to the α subunit of the high affinity IgE receptor and the IgE molecule are proposed to be of pathogenetic relevance in a group of patients with chronic urticaria (CU). The diagnosis of autoimmune chronic urticaria (ACU) is difficult; the autologous serum skin test (ASST) seems to be a useful screening test, but reliable, additional confirmatory methods are needed. Objectives: To assess the diagnostic value of a modified serum-induced basophil activation test, the CD63 expression assay, in the diagnosis of ACU by comparing the results of the CD63 assay with the results of the histamine release (HR) test, the ASST and serum levels of soluble CD40 ligand (sCD40L). Methods: Using basophils from an atopic (DA) and a nonatopic (DNA) donor the activity of sera of 72 patientswith CU were measured in HR assay by enzyme-linked immunosorbent assay and in CD63 expression assay by flow cytometry. An ASST was carried out in all patients; in 30 of the 72 patients sCD40L was detected and correlationswere derived between the different assays. Sera of 20 normal controls and 26 patients with systemic autoimmune diseases were also tested in the HR assay and in the CD63 expression assay. Results: Histamine-releasing activity was detected in the sera of 51% (DA) and 32% (DNA) of CU patients and 57% (DA) and 28% (DNA) of sera upregulated CD63 expression on the surface of basophils from the different donors. There was a significant correlation between the HR and the CD63 assays carried out on both donors, but the ASST showed a strong correlation with the HR assay only for basophils from the DA. The serum level of sCD40L was significantly higher in patients with CU compared with controls, but the difference between the autoimmune and the nonautoimmune groups was not significant. Conclusions: The CD63 expression assay seems to be a reliable functional test in the diagnosis of ACU, particularly if highly sensitive donor basophils are used, but the determination of the sCD40L serum level was not sufficient to differentiate between the autoimmune and the nonautoimmune patient groups.展开更多
文摘Background: The autoimmune subclass of chronic idiopathic urticaria (CU) has been characterized by the occurrence of biologically relevant IgG antibodies against the IgE molecule or the αchain of the high-affinity Fc receptor (FcRIα) on basophils and mast cells. These antibodies are usually detected by autologous serum skin testing and confirmed by histamine release studies, immunoblotting, or enzyme-linked immunosorbent assay, but not always. Objectives: To detect autoantibodies to the FcRIαin sera of CU patients by a modified serum-induced basophil activation testmeasured by flowcytometry (FCM) and to evaluate the relationship between the in vitro functional test, the autologous serum skin test (ASST), and the serum levels of IgE, eosinophil cationic protein (ECP) and antithyroid antibodies. Methods: Sera of 30 patients with CU and 26 patients with systemic autoimmune diseases (systemic lupus erythematosus, dermatomyositis) were tested for CD63 activation marker expression on basophils by FCM. Leucocytes from two highly sensitized atopic donors (DA1. DA2) and one non-atopic donor (DNA) were incubated with patients’sera and double-labelled with anti-IgE and anti-CD63 antibodies. Subsequently, the percentage of CD63-expressing basophils was determined by using FCM. In all CU patients an ASST was carried out and the serum IgE, and ECP levels and antithyroid antibodies were evaluated. Results: Twelve patients had a positive ASST and 14 patients a positive CD63 expression assay. There was a strong correlation between the ASST and CD63 assay. Sera from patients with systemic autoimmune diseases did not raise positive CD63 expression on basophils. There was a moderate negative correlation between the occurrence of atopic serum markers (IgE, ECP) and the ability of sera to induce CD63 expression on basophil cells of DA2 (P < 0.05). The female sex was preponderant and antithyroid antibodies were more frequent. Conclusions: Our new technical observation demonstrates that basophils of highly sensitized atopic donors can be successfully used without priming with IL-3 for the in-vitro flow cytofluorimetric diagnosis of CU. With this investigation the characterization of the autoimmune origin of CU is based on an objective in vitro technique.
文摘Background: Antibodies directed to the α subunit of the high affinity IgE receptor and the IgE molecule are proposed to be of pathogenetic relevance in a group of patients with chronic urticaria (CU). The diagnosis of autoimmune chronic urticaria (ACU) is difficult; the autologous serum skin test (ASST) seems to be a useful screening test, but reliable, additional confirmatory methods are needed. Objectives: To assess the diagnostic value of a modified serum-induced basophil activation test, the CD63 expression assay, in the diagnosis of ACU by comparing the results of the CD63 assay with the results of the histamine release (HR) test, the ASST and serum levels of soluble CD40 ligand (sCD40L). Methods: Using basophils from an atopic (DA) and a nonatopic (DNA) donor the activity of sera of 72 patientswith CU were measured in HR assay by enzyme-linked immunosorbent assay and in CD63 expression assay by flow cytometry. An ASST was carried out in all patients; in 30 of the 72 patients sCD40L was detected and correlationswere derived between the different assays. Sera of 20 normal controls and 26 patients with systemic autoimmune diseases were also tested in the HR assay and in the CD63 expression assay. Results: Histamine-releasing activity was detected in the sera of 51% (DA) and 32% (DNA) of CU patients and 57% (DA) and 28% (DNA) of sera upregulated CD63 expression on the surface of basophils from the different donors. There was a significant correlation between the HR and the CD63 assays carried out on both donors, but the ASST showed a strong correlation with the HR assay only for basophils from the DA. The serum level of sCD40L was significantly higher in patients with CU compared with controls, but the difference between the autoimmune and the nonautoimmune groups was not significant. Conclusions: The CD63 expression assay seems to be a reliable functional test in the diagnosis of ACU, particularly if highly sensitive donor basophils are used, but the determination of the sCD40L serum level was not sufficient to differentiate between the autoimmune and the nonautoimmune patient groups.
