Characterization of surface polypeptides on different life-cycle stages of Theileria annulata

Abstract

We describe the characterisation of polypeptides located on the surface of Theileria annulata sporozoites, macroschizonts, piroplasms and infected lymphoblastoid cells using surface iodination techniques. The sporozoite stage exhibited a complex profile of surface polypeptides. However, using data from experiments with defined monoclonal antibodies, the sporozoite surface appeared to be composed of several distinct groups of related polypeptides. Analysis of the macroschizont detected seven surface polypeptides, while eight polypeptides were identified for the piroplasm stage. On the basis of molecular weight comparisons, one of the surface polypeptides appeared to be common to the sporozoite, macroschizont and piroplasm. Stage cross-reactive monoclonals failed to immunoprecipitate a surface-radiolabelled polypeptide, and this prohibited the characterisation of a stage common surface antigen. From the surface labelling studies of Theileria-infected and uninfected lymphoblastoid cell lines, we concluded that infection results in major changes at the surface of the host cell, including both the appearance and loss of specific polypeptides. By employing monoclonal antibodies which detect infection-associated determinants, and a polyclonal antiserum raised against a glycoprotein fraction of an infected cell lysate, surface-labelled polypeptides were specifically immunoprecipitated from extracts of infected cells. The polypeptide detected by monoclonal antibody 4H5 was characterised as an infection-associated glycoprotein which varies in molecular mass when immunoprecipitated from different infected cell lines. The identification of infection-associated glycoproteins on the surface of the lymphoblastoid cell suggests that these molecules may be recognised by the cytotoxic T cells of immune animals.