Isolation and characterization of a monoclonal anti-P30 antibody resistant mutant of Toxoplasma gondii.
Of the possible iodine-labelled Toxoplasma gondii surface proteins, P30 (apparent Mr 30,000) is the principal one recognized by acute and convalescent anti-toxoplasma sera. This protein which comprises from 3 to 5% of the total parasite protein was used to raise a panel of parasiticidal monoclonal anti-P30 antibodies. One of these monoclonal antibodies was able to select a resistant mutant from a large population of chemically mutagenized wild-type P strain parasites. This mutant retained the wild type sensitivity to other non-P30 parasiticidal monoclonal antibodies as well as polyclonal anti-P30 rabbit sera. Analysis of surface radioiodinated wild type and mutant parasites showed that the mutant had a quantitative reduction in the amount of P30. A comparison of surface biotin labelled wild type and resistant parasites by two dimensional electrophoresis showed that the mutant lacked one and possibly two of several proteins that make up wild type P30. Western blot analysis indicated that the mutant was devoid of antigenically reactive P30. These findings further support the hypothesis that antigenic variants of T. gondii can be induced and may involve the major surface membrane antigens of the parasite.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000953:Antigens, Protozoan / Q000032:analysis* / Q000276:immunology; D000954:Antigens, Surface / Q000032:analysis / Q000276:immunology; D001345:Autoradiography; D004591:Electrophoresis, Polyacrylamide Gel; D007118:Immunoassay; D008565:Membrane Proteins / Q000032:analysis* / Q000276:immunology; D009154:Mutation; D015800:Protozoan Proteins; D014122:Toxoplasma / Q000032:analysis / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan; D000954:Antigens, Surface; D008565:Membrane Proteins; D015800:Protozoan Proteins; C047168:SAG1 antigen, Toxoplasma