title: Identification of Antigenic Proteins from Lichtheimia corymbifera for Farmer's Lung Disease Diagnosis.
abstract: The use of recombinant antigens has been shown to improve both the sensitivity and the standardization of the serological diagnosis of Farmer's lung disease (FLD). The aim of this study was to complete the panel of recombinant antigens available for FLD serodiagnosis with antigens of Lichtheimia corymbifera, known to be involved in FLD. L. corymbifera proteins were thus separated by 2D electrophoresis and subjected to western blotting with sera from 7 patients with FLD and 9 healthy exposed controls (HEC). FLD-associated immunoreactive proteins were identified by mass spectrometry based on a protein database specifically created for this study and subsequently produced as recombinant antigens. The ability of recombinant antigens to discriminate patients with FLD from controls was assessed by ELISA performed with sera from FLD patients (n = 41) and controls (n = 43) recruited from five university hospital pneumology departments of France and Switzerland. Forty-one FLD-associated immunoreactive proteins from L. corymbifera were identified. Six of them were produced as recombinant antigens. With a sensitivity and specificity of 81.4 and 77.3% respectively, dihydrolipoyl dehydrogenase was the most effective antigen for discriminating FLD patients from HEC. ELISA performed with the putative proteasome subunit alpha type as an antigen was especially specific (88.6%) and could thus be used for FLD confirmation. The production of recombinant antigens from L. corymbifera represents an additional step towards the development of a standardized ELISA kit for FLD diagnosis.
authors.affiliation: UMR6249 Chrono-Environnement, University of Bourgogne Franche-Comté, Besançon, France.|Parasitology-Mycology Department, University Hospital of Besançon, Besançon, France.
authors.forename: Bénédicte
authors.initials: B
authors.lastname: Rognon
authors.affiliation: UMR6249 Chrono-Environnement, University of Bourgogne Franche-Comté, Besançon, France.|Parasitology-Mycology Department, University Hospital of Besançon, Besançon, France.
authors.forename: Coralie
authors.initials: C
authors.lastname: Barrera
authors.affiliation: Department of Dermatology, University Hospital, Lausanne, Switzerland.
authors.forename: Michel
authors.initials: M
authors.lastname: Monod
authors.affiliation: UMR6249 Chrono-Environnement, University of Bourgogne Franche-Comté, Besançon, France.|Parasitology-Mycology Department, University Hospital of Besançon, Besançon, France.
authors.forename: Benoit
authors.initials: B
authors.lastname: Valot
authors.affiliation: UMR6249 Chrono-Environnement, University of Bourgogne Franche-Comté, Besançon, France.|Parasitology-Mycology Department, University Hospital of Besançon, Besançon, France.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Roussel
authors.affiliation: Protein Analysis Facility, Center for Integrative Genomics, University of Lausanne, Lausanne, Switzerland.
authors.forename: Manfredo
authors.initials: M
authors.lastname: Quadroni
authors.affiliation: Pneumology Department, University Hospital of Rennes, Rennes, France.|UMR1085, IRSET, Rennes 1 University, Rennes, France.
authors.forename: Stephane
authors.initials: S
authors.lastname: Jouneau
authors.affiliation: Pneumology Department, University Hospital North, Saint-Etienne, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Court-Fortune
authors.affiliation: Pneumology Department, University Hospital Gabriel Montpied, Clermont-Ferrand, France.
authors.forename: Denis
authors.initials: D
authors.lastname: Caillaud
authors.affiliation: Department of Pulmonary Medicine, Pourtalès Hospital, Neuchatel, Switzerland.
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Fellrath
authors.affiliation: UMR6249 Chrono-Environnement, University of Bourgogne Franche-Comté, Besançon, France.|Department of Respiratory Disease, University Hospital of Besançon, Besançon, France.
authors.forename: Jean-Charles
authors.initials: JC
authors.lastname: Dalphin
authors.affiliation: UMR6249 Chrono-Environnement, University of Bourgogne Franche-Comté, Besançon, France.|Parasitology-Mycology Department, University Hospital of Besançon, Besançon, France.
authors.forename: Gabriel
authors.initials: G
authors.lastname: Reboux
authors.affiliation: UMR6249 Chrono-Environnement, University of Bourgogne Franche-Comté, Besançon, France.|Parasitology-Mycology Department, University Hospital of Besançon, Besançon, France.
authors.forename: Laurence
authors.initials: L
authors.lastname: Millon
mesh_terms: D000946:Antigens, Fungal / Q000145:classification / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D015153:Blotting, Western; D016022:Case-Control Studies; D030541:Databases, Genetic; D015180:Electrophoresis, Gel, Two-Dimensional; D004797:Enzyme-Linked Immunosorbent Assay; D005203:Farmer's Lung / Q000175:diagnosis* / Q000382:microbiology; D005260:Female; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D008297:Male; D013058:Mass Spectrometry; D009090:Mucorales / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D012331:RNA, Fungal / Q000302:isolation & purification / Q000378:metabolism; D012333:RNA, Messenger / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D017423:Sequence Analysis, RNA
chemical_list: D000946:Antigens, Fungal; D007074:Immunoglobulin G; D012331:RNA, Fungal; D012333:RNA, Messenger; D011994:Recombinant Proteins