title: Monoclonal antibodies to hyphal exoantigens derived from the opportunistic pathogen Aspergillus terreus.
abstract: Aspergillus terreus has been difficult to identify in cases of aspergillosis, and clinical identification has been restricted to the broad identification of aspergillosis lesions in affected organs or the detection of fungal carbohydrates. As a result, there is a clinical need to identify species-specific biomarkers that can be used to detect invasive A. terreus disease. Monoclonal antibodies (MAbs) were developed to a partially purified preparation of cytolytic hyphal exoantigens (HEA) derived from A. terreus culture supernatant (CSN). Twenty-three IgG1 isotype murine MAbs were developed and tested for cross-reactivity against hyphal extracts of 54 fungal species. Sixteen MAbs were shown to be specific for A. terreus. HEA were detected in conidia, hyphae, and in CSN of A. terreus. HEA were expressed in high levels in the hyphae during early stages of A. terreus growth at 37°C, whereas at room temperature the expression of HEA peaked by days 4 to 5. Expression kinetics of HEA in CSN showed a lag, with peak levels at later time points at room temperature and 37°C than in hyphal extracts. Serum spiking experiments demonstrated that human serum components do not inhibit detection of the HEA epitopes by MAb enzyme-linked immunosorbent assay (ELISA). Immunoprecipitation and proteomic analysis demonstrated that MAbs 13E11 and 12C4 immunoprecipitated a putative uncharacterized leucine aminopeptidase (Q0CAZ7), while MAb 19B2 recognized a putative dipeptidyl-peptidase V (DPP5). Studies using confocal laser scanning microscopy showed that the uncharacterized leucine aminopeptidase mostly localized to extracellular matrix structures while dipeptidyl-peptidase V was mostly confined to the cytoplasm.
authors: Allergy and Clinical Immunology Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Centers for Disease Control and Prevention, 1095 Willowdale Road, Morgantown, WV 26505, USA.
authors: Ajay P
authors: AP
authors: Nayak
authors: Brett J
authors: BJ
authors: Green
authors: Erika
authors: E
authors: Janotka
authors: Justin M
authors: JM
authors: Hettick
authors: Sherri
authors: S
authors: Friend
authors: Steve J
authors: SJ
authors: Vesper
authors: Detlef
authors: D
authors: Schmechel
authors: Donald H
authors: DH
authors: Beezhold
mesh_terms: D000818:Animals; D000908:Antibodies, Fungal / Q000096:biosynthesis / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology; D000946:Antigens, Fungal / Q000032:analysis / Q000276:immunology*; D001228:Aspergillosis / Q000175:diagnosis* / Q000276:immunology / Q000382:microbiology; D001230:Aspergillus / Q000145:classification / Q000276:immunology*; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / Q000032:analysis / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D025301:Hyphae / Q000276:immunology*; D047468:Immunoprecipitation; D007931:Leucyl Aminopeptidase / Q000032:analysis / Q000276:immunology; D051379:Mice; D018613:Microscopy, Confocal; D009894:Opportunistic Infections / Q000175:diagnosis* / Q000276:immunology / Q000382:microbiology; D040901:Proteomics; D011817:Rabbits; D013045:Species Specificity
chemical_list: D000908:Antibodies, Fungal; D000911:Antibodies, Monoclonal; D000946:Antigens, Fungal; D007931:Leucyl Aminopeptidase; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases