PRODUCTION OF ANTIIDIOTYPIC MONOCLONAL-ANTIBODIES THAT MIMIC THE PHYTOTOXIN DOTHISTROMIN

Dothistromin (DOTH) is a toxin secreted by Dothistroma pini and is pre sent in needle lesions resulting from infection by this fungus. A mono clonal antibody (MAb 10C12) of high affinity for DOTH was conjugated t o bovine serum albumin (BSA) and used to immunize mice in order to pre pare monoclonal anti-idiotypic antibody mimics of dothistromin. One an ti-idiotypic antibody, 12C9G8, was equivalent to DOTH on a molar basis in inhibition of the binding of an anti-DOTH MAb to DOTH-protein conj ugates. Furthermore, in a Western blot analysis, 12C9G8 visualized the same protein profile in embryos as was observed in an approach which utilized an anti-DOTH MAb together with DOTH-mouse serum albumin (MSA) conjugate to identify potential DOTH binding sites. In a parallel imm unogold electron microscopy experiment, protein-containing vesicles we re labelled using both approaches.