Characterisation of bovine transferrin receptor on normal activated and Theileria parva-transformed lymphocytes by a new monoclonal antibody

A murine IgM monoclonal antibody (mAb), IL-A77, has been generated that recognises the bovine transferrin receptor (TfR) and will be a useful tool to measure the activation state of bovine lymphocytes and macrophages. The antigen is detected on immature erythroid cells and proliferating lymphocytes. It is undetectable on resting lymphocytes, but appears within 24 h after stimulation with concanavalin A (ConA) or pokeweed mitogen (PWM). Immune precipitations of lysates of both labeled activated lymphocytes and bone marrow erythroid cells showed that, similar to human TfR, the bovine receptor is a disulfide-bonded dimer of two identical chains of M 97 000. A similar 97000 M protein was eluted from a column containing immobilised bovine transferrin (Tf) using conditions known to elute the human TfR, and this protein was recognised by mAb IL-A77, proving that it detected bovine TfR. Although the mAb inhibited binding of transferrin to its receptor, it did not block proliferation of Theileria parva-transformed or ConA-stimulated lymphocytes. When cells were metabolically labeled with 35 S-methionine, a second 90 000-M TfR band was detected in Theileria parva-transformed cells, but not in stimulated lymphocytes. This form of the TfR was not expressed on the cell surface. It may be an usual precursor of the receptor, a parasite-modified receptor or it may be of parasite origin and necessary for transfer of iron into the intracellular parasite