Phenotypic variation and diversity among Trichomonas vaginalis isolates and correlation of phenotype with trichomonal virulence determinants

Abstract

The extent and nature of heterogeneity among representative Trichomonas vaginalis isolates were evaluated by flow cytofluorometry analysis. Monoclonal antibody and monospecific antiserum to an immunodominant trichomonad surface glycoprotein with a molecular weight of 267,000 (267K glycoprotein) were used to evaluate fresh isolates (JHH and RU375) and long-term grown isolates (NYH286, IR78, and JH31A) of T. vaginalis. Isolates NYH286, JH31A, and JHH were made up of heterogeneous staining (positive [pos] phenotype) and nonstaining (negative [neg] phenotype) populations of trichomonads, whereas RU375 and IR78 were all neg phenotype parasites. Flow cytofluorometric patterns of agar clones derived from single organisms of heterogeneous isolates such as NYH286 showed populations which were either homogeneous pos or neg and also showed clones which were heterogeneous in nature containing both phenotypes. Fluorescence-activated cell sorting was also accomplished, and subpopulations of defined pos or neg phenotype were purified. Flow cytofluorometry evaluation of all isolates for an extended period revealed a phenotypic variation among all heterogeneous isolates and also for all clones and subpopulations derived from the heterogeneous isolates. On the other hand, IR78 and RU375 did not undergo phenotypic variation and have remained neg for greater than 4 years. Parasites which were nonreactive with either monoclonal antibody or monospecific antiserum to the 267K glycoprotein in flow cytofluorometry did not possess the antigen on their surface. This was determined by radioimmunoprecipitation assays using extracts of iodinated trichomonads. Finally, neg phenotype parasites yielded enhanced rates of contact-dependent cytotoxicity of host cell monolayers as compared with the pos phenotype trichomonads.