Monoclonal antibody to a major surface glycoprotein immunogen differentiates isolates and subpopulations of Trichomonas vaginalis.
To produce monoclonal antibodies (MAbs) to highly immunogenic membrane proteins of Trichomonas vaginalis NYH286, the sera of subcutaneously infected BALB/c mice were first monitored for antibody to trichomonad surface proteins. The sera possessed antibody to one major surface protein by 7 days and antibody to numerous other trichomonad membrane proteins by 4 weeks postinfection. A hybridoma was then generated that synthesized an MAb, designated C20A3, which reacted to a parasite-derived glycoprotein possessing a molecular weight of 267,000 (267K glycoprotein). The immunogen corresponded to the single high-molecular-weight immunogenic surface protein recognized by 7-day mouse antisera. The MAb differentiated T. vaginalis isolates by a whole-cell enzyme-linked immunosorbent assay and by indirect immunofluorescence, using either fixed or live organisms. All isolates, however, possessed C20A3-reactive material when tested by enzyme-linked immunosorbent assay, using detergent extracts of the isolates incubated with MAb-coated microtiter well plates. The epitope was accessible to antibody binding on live T. vaginalis organisms expressing the major immunogen, and the 267K glycoprotein was readily removed from the parasite membranes by trypsinizing the intact trichomonads. The antigen incorporated radiolabeled glucose, mannose, and acetate. Also, an unlabeled 267K glycoprotein on nitrocellulose blots was detected by 125I-concanavalin A and 125I-wheat germ agglutinin, confirming the glycoprotein nature of the immunogen. Finally, of seven isolates used in this study, one possessed a cross-reactive 170K, rather than 267K, antigen. The data reinforce the idea that antigenic heterogeneity among T. vaginalis isolates may be a function of the presence or absence of high-molecular-weight glycoprotein immunogens from trichomonal membranes.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000917:Antibody Formation; D000953:Antigens, Protozoan / Q000008:administration & dosage / Q000276:immunology*; D006023:Glycoproteins / Q000276:immunology; D006801:Humans; D007163:Immunosorbent Techniques; D051379:Mice; D008970:Molecular Weight; D014245:Trichomonas Infections / Q000276:immunology*; D014246:Trichomonas vaginalis / Q000145:classification / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan; D006023:Glycoproteins