Immunological and biochemical analysis of glycosylated surface antigens and lipophosphoglycan of Tritrichomonas foetus.
Immunoaffinity-purified TF1.17 adhesin antigen was compared biochemically and antigenically to Tritrichomonas foetus (TF) lipophosphoglycan (LPG) and a soluble glycosylated antigen (SGA) released from T. foetus and implicated in pathogenesis and immunity. The monoclonal antibodies (Mabs TF1.15 and TF1.17) specific for a glycosylated TF1.17 antigen were previously shown to prevent adhesion of the T. foetus parasites to bovine vaginal epithelial cells and to mediate killing by bovine complement. SGA was isolated from T. foetus-conditioned buffer and purified by octyl-Sepharose hydrophobic column chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of SGA showed a major SGA1 component (approximately 190 kDa) and a minor SGA2 component (50-70 kDa), which migrated close to TF-LPG and TF1.17. The carbohydrate and lipid compositional analyses of affinity-purified TF1.17 and SGA2 by high-performance liquid chromatography (HPLC) and gas-liquid chromatography revealed the presence of monosaccharides and fatty acids as found in TF-LPG. All antigens contained terminal fucose as determined by alpha-fucosidase digestion followed by HPLC. ELISA and western blots were used to further characterize these glycosylated antigens and to analyze their relationships. The Mabs TF1.15 and TF1.17 reacted very strongly to TF-LPG and SGA2. as well as TF1.17 antigen, indicating that these molecules share common epitopes. These Mabs did not react with the SGA1 component either in ELISA and western blot analyses. Also, the monosaccharide composition of SGA1 was very different from the other three antigen, suggesting SGA1 was different from LPG, SGA2 and TF1.17. Although LPG reacted with Mabs to native TF1.17 antigen, LPG did not induce an immune response in cattle with the same route and adjuvant used to produce strong antibody responses to the native antigen. The latter response suggests that the tightly bound peptide present in the immunoaffinity-purified antigen is necessary for induction of a response to (an) epitope(s) in TF-LPG and TF1.17. Furthermore, vaginal fluid from T. foetus-infected heifers and serum from a cow with a T. foetus-associated pyometra recognized both TF1.17 and TF-LPG in western blots. These results suggest that T. foetus LPG and SGA2 are related to TF1.17 antigen, which was previously shown to play an important role in the pathogenesis and host response in bovine trichomoniasis.
Singh
B N
BN
Department of Biochemistry and Molecular Biology, SUNY UpState Medical University, Syracuse, New York 13210, USA.
BonDurant
R H
RH
Campero
C M
CM
Corbeil
L B
LB
D000818:Animals; D000953:Antigens, Protozoan / Q000737:chemistry* / Q000276:immunology; D000954:Antigens, Surface / Q000737:chemistry* / Q000276:immunology; D002417:Cattle; D002418:Cattle Diseases / Q000209:etiology; D005227:Fatty Acids / Q000032:analysis; D005260:Female; D006023:Glycoproteins / Q000737:chemistry* / Q000276:immunology; D006028:Glycosphingolipids / Q000737:chemistry* / Q000276:immunology; D006031:Glycosylation; D009005:Monosaccharides / Q000032:analysis; D011528:Protozoan Infections / Q000209:etiology; D011529:Protozoan Infections, Animal; D012749:Sexually Transmitted Diseases / Q000453:epidemiology / Q000662:veterinary; D012995:Solubility; D016835:Tritrichomonas foetus / Q000276:immunology*; D014611:Vaccination
D000953:Antigens, Protozoan; D000954:Antigens, Surface; D005227:Fatty Acids; D006023:Glycoproteins; D006028:Glycosphingolipids; D009005:Monosaccharides; C008290:lipophosphonoglycan