Immuno-reactive proteins from Mycobacterium immunogenum useful for serodiagnosis of metalworking fluid hypersensitivity pneumonitis.
Metalworking fluid-associated hypersensitivity pneumonitis (MWF-HP) is a pulmonary disease caused by inhaling microorganisms present in the metalworking fluids used in the industrial sector. Mycobacterium immunogenum is the main etiological agent. Among the clinical, radiological and biological tools used for diagnosis, serological tests are important. The aim of this study was to identify immunogenic proteins in M. immunogenum and to use recombinant antigens for serological diagnosis of MWF-HP. Immunogenic proteins were detected by two-dimensional Western blot and candidate proteins were identified by mass spectrometry. Recombinant antigens were expressed in Escherichia coli and tested by enzyme-linked immunosorbent assay (ELISA) with the sera of 14 subjects with MWF-HP and 12 asymptomatic controls exposed to M. immunogenum. From the 350 spots visualized by two-dimensional gel electrophoresis with M. immunogenum extract, 6 immunogenic proteins were selected to be expressed as recombinant antigens. Acyl-CoA dehydrogenase antigen allowed for the best discrimination of MWF-HP cases against controls with an area under the receiver operating characteristics (ROC) curve of 0.930 (95% CI=0.820-1), a sensitivity of 100% and a specificity of 83% for the optimum threshold. Other recombinant antigens correspond to acyl-CoA dehydrogenase FadE, cytosol aminopeptidase, dihydrolipoyl dehydrogenase, serine hydroxymethyltransferase and superoxide dismutase. This is the first time that recombinant antigens have been used for the serodiagnosis of hypersensitivity pneumonitis. The availability of recombinant antigens makes it possible to develop standardized serological tests which in turn could simplify diagnosis, thus making it less invasive.
Roussel
Sandrine
S
UMR/CNRS Chrono-Environnement 6249, University of Franche-Comté, France.
Rognon
Bénédicte
B
Barrera
Coralie
C
Reboux
Gabriel
G
Salamin
Karine
K
Grenouillet
Frédéric
F
Thaon
Isabelle
I
Dalphin
Jean-Charles
JC
Tillie-Leblond
Isabelle
I
Quadroni
Manfredo
M
Monod
Michel
M
Millon
Laurence
L
D000542:Alveolitis, Extrinsic Allergic / Q000175:diagnosis*; D000942:Antigens, Bacterial / Q000097:blood*; D001426:Bacterial Proteins / Q000032:analysis / Q000276:immunology; D019411:Clinical Laboratory Techniques / Q000379:methods*; D003001:Cloning, Molecular; D015180:Electrophoresis, Gel, Two-Dimensional; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D009161:Mycobacterium / Q000276:immunology*; D009784:Occupational Diseases / Q000175:diagnosis*; D020543:Proteome / Q000032:analysis; D012372:ROC Curve; D011994:Recombinant Proteins / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D020543:Proteome; D011994:Recombinant Proteins