Production and immunological characterization of a monoclonal antibody to Trichophyton quinckeanum: interaction with phosphorylcholine-bearing components.
A monoclonal antibody (Tq-1) that interacts with the phosphorylcholine (PC)-bearing antigens of Trichophyton quinckeanum was produced by fusion of myeloma cells (JKAg-8) with spleen cells of BALB/c mice immunized with an alum-precipitated fraction of T. quinckeanum cytoplasmic antigen. It was characterized as an IgM class antibody by immunodiffusion using anti-Ig heavy chain specific reagents, ELISA using immunoglobulin-specific peroxidase-conjugated antibodies, and by gel filtration chromatography; it showed high affinity for Staphylococcus aureus protein-A. Interaction of Tq-1 with PC-like antigens of T. quinckeanum was demonstrated by inhibition studies using ELISA, immunoblotting, immunoprecipitation and immuno electron microscopy techniques. The binding activity of Tq-1 antibody with a range of dermatophyte proteins was completely inhibited by prior incubation with PC hapten. Moreover, dermatophyte antigens reacting with the monoclonal antibody reacted strongly with sera from chronically infected mice. Dermatophyte antigens derived from both young (24 h) and old (20 d) cultures reacted with Tq-1 and this binding was inhibited by PC, suggesting that Tq-1 target antigen PC appears at an early stage during fungal growth and remains throughout its life.
Calderon
R A
RA
Department of Medical Microbiology, London School of Hygiene and Tropical Medicine, UK.
Shennan
G I
GI
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D000946:Antigens, Fungal / Q000032:analysis; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology*; D002794:Choline / Q000031:analogs & derivatives*; D004797:Enzyme-Linked Immunosorbent Assay; D006825:Hybridomas; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron; D010767:Phosphorylcholine / Q000276:immunology*; D018272:Porins; D014249:Trichophyton / Q000276:immunology* / Q000648:ultrastructure
D000911:Antibodies, Monoclonal; D000946:Antigens, Fungal; D001425:Bacterial Outer Membrane Proteins; D018272:Porins; C057699:porin protein, Neisseria; D010767:Phosphorylcholine; D002794:Choline