title: Negative immunodiffusion test results obtained with sera of paracoccidioidomycosis patients may be related to low-avidity immunoglobulin G2 antibodies directed against carbohydrate epitopes.
abstract: Immunodiffusion (ID) is the serologic test most frequently used for the diagnosis and posttherapy follow-up of patients with paracoccidioidomycosis (PCM). The ID test is highly specific (100%), but its sensitivity is relatively low (90%), leading to false-negative results. The aim of this study was to determine the profiles of antibodies in sera from patients with proven PCM and with negative results in the ID test (IDneg) versus positive results in the ID test (IDpos). We analyzed 46 sera from patients with active PCM for total immunoglobulin G (IgG) and IgG subclass responses to Paracoccidioides brasiliensis gp43 antigen (treated or not treated with sodium metaperiodate) by enzyme-linked immunosorbent assay and immunoblotting. Immunoblotting showed that both IDneg and IDpos sera recognized predominantly the gp43 fraction of the P. brasiliensis antigen used in the ID test. IDneg sera contain low-avidity antibodies, low levels of specific IgG (total) and IgG1, and high levels of IgG2 compared with IDpos sera. The antibodies present in IDneg sera were predominantly directed against carbohydrate epitopes, since treatment with sodium metaperiodate resulted in a significant decrease in antibody reactivity. These data suggest that the lack of reactivity of sera from PCM patients in the ID test may be related to the production of low-avidity IgG2 antibodies directed against carbohydrate epitopes.
authors.affiliation: Department of Clinical Pathology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), Campinas, Brazil.
authors.forename: Andréia R
authors.initials: AR
authors.lastname: Neves
authors.forename: Ronei L
authors.initials: RL
authors.lastname: Mamoni
authors.forename: Cláudio L
authors.initials: CL
authors.lastname: Rossi
authors.forename: Zoilo P
authors.initials: ZP
authors.lastname: de Camargo
authors.forename: Maria Heloísa S L
authors.initials: MH
authors.lastname: Blotta
mesh_terms: D000818:Animals; D000908:Antibodies, Fungal / Q000097:blood / Q000276:immunology*; D000946:Antigens, Fungal / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005656:Fungal Proteins / Q000276:immunology; D006023:Glycoproteins / Q000276:immunology; D006801:Humans; D015151:Immunoblotting; D005779:Immunodiffusion / Q000379:methods; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D007132:Immunoglobulin Isotypes; D010228:Paracoccidioides / Q000276:immunology; D010229:Paracoccidioidomycosis / Q000175:diagnosis* / Q000276:immunology*; D012680:Sensitivity and Specificity
chemical_list: C103180:43 kDa protein, Paracoccidioides; D000908:Antibodies, Fungal; D000946:Antigens, Fungal; D005656:Fungal Proteins; D006023:Glycoproteins; D007074:Immunoglobulin G; D007132:Immunoglobulin Isotypes