title: Degradation of bovine complement C3 by trichomonad extracellular proteinase.
abstract: Bovine trichomoniasis is a local infection of the reproductive tract making interaction with mucosal host defenses crucial. Since the parasite is susceptible to killing by bovine complement, we investigated the role of the third component of complement (C3) in host parasite interactions. Bovine C3 was purified by anionic and cationic exchange chromatography. The purified protein was characterized by immunoreactivity, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and peptide sequencing of the amino terminus of the beta chain. When purified bovine C3 was incubated for varying time periods with trichomonad extracellular proteinases, SDS-PAGE gels revealed digestion of the alpha chain to small fragments. Such degradation in vivo would prevent formation of C3b and completion of the complement cascade, resulting in evasion of killing. To evaluate the relevance of this data, we determined whether C3 was present in bovine genital secretions. With a quantitative ELISA assay, C3 could be demonstrated in both uterine and vaginal washes. To our knowledge, this is the first demonstration of bovine C3 in genital secretions. The C3 concentration increased significantly in vaginal secretions by 8 and 10 weeks in heifers infected with Tritrichomonas foetus. An increase was also seen in uterine secretions of infected heifers, but sample numbers were insufficient for statistical analysis. Transcription of the major extracellular cysteine proteinase (TFCP8) was demonstrated in T. foetus cells from uterine secretions of infected heifers by RT-PCR and Southern blotting. The results indicate that C3 may be important in genital defense and that trichomonad extracellular proteinases may play a role in evasion of complement-mediated killing.
authors.affiliation: Department of Comparative Medicine, University of Tennessee, Knoxville, TN 37901-1071, USA.
authors.forename: S A
authors.initials: SA
authors.lastname: Kania
authors.forename: S L
authors.initials: SL
authors.lastname: Reed
authors.forename: J W
authors.initials: JW
authors.lastname: Thomford
authors.forename: R H
authors.initials: RH
authors.lastname: BonDurant
authors.forename: K
authors.initials: K
authors.lastname: Hirata
authors.forename: R R
authors.initials: RR
authors.lastname: Corbeil
authors.forename: M J
authors.initials: MJ
authors.lastname: North
authors.forename: L B
authors.initials: LB
authors.lastname: Corbeil
mesh_terms: D000818:Animals; D015139:Blotting, Southern / Q000662:veterinary; D002417:Cattle; D002418:Cattle Diseases / Q000276:immunology* / Q000378:metabolism / Q000469:parasitology; D003176:Complement C3 / Q000276:immunology / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005260:Female; D011528:Protozoan Infections / Q000276:immunology* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016053:RNA, Protozoan / Q000737:chemistry / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D016835:Tritrichomonas foetus / Q000201:enzymology*; D014599:Uterus / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D014621:Vagina / Q000276:immunology / Q000378:metabolism / Q000469:parasitology
chemical_list: D003176:Complement C3; D012333:RNA, Messenger; D016053:RNA, Protozoan; D003546:Cysteine Endopeptidases