title: Immunization with a tubulin-rich preparation from Trypanosoma brucei confers broad protection against African trypanosomosis.
abstract: Tubulin from Trypanosoma brucei was purified to near homogeneity using a protocol which involved treatment with urea with subsequent renaturation and was then used to immunize mice. Renatured tubulin further purified by SDS-PAGE (denatured), synthetic tubulin peptides (STP), and rat brain tubulin (RbTub) were also used. Immunized mice were challenged with T. brucei, Trypanosoma congolense or Trypanosoma rhodesiense. Renatured T. brucei tubulin (nTbTub) induced protection in all mice tested, of which 60-80% (n = 81) was complete and the remainder partial. Denatured T. brucei tubulin (dTbTub), STP, or RbTub induced lower antibody levels than nTbTub and did not offer protection. However, in culture, the antibodies against dTbTub or STP killed trypanosomes although at lower dilutions than nTbTub, but those against RbTub did not. In Western blots anti-trypanosome antibodies recognized the tubulin of all the trypanosome species investigated but not vertebrate tubulin, whereas the anti-RbTUB antibodies recognized both trypanosome and vertebrate tubulin. Of the five mice given passive immunity by the transfer of anti-nTbTub serum, four were completely protected and one partially protected. These data suggest that tubulin is the relevant immunogen in the preparation used and could therefore be a promising target for the development of a parasite-specific, broad spectrum vaccine.
authors.affiliation: Molecular Biology Laboratory, Faculty of Veterinary Medicine, Makerere University, PO Box 7062, Kampala, Uganda. glubega@imul.com
authors.forename: George W
authors.initials: GW
authors.lastname: Lubega
authors.forename: Denis K
authors.initials: DK
authors.lastname: Byarugaba
authors.forename: Roger K
authors.initials: RK
authors.lastname: Prichard
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000096:biosynthesis / Q000097:blood; D000918:Antibody Specificity; D000953:Antigens, Protozoan / Q000276:immunology / Q000302:isolation & purification; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D007106:Immune Sera / Q000276:immunology; D007114:Immunization; D007116:Immunization, Passive; D007274:Injections, Intraperitoneal; D007279:Injections, Subcutaneous; D051379:Mice; D010455:Peptides / Q000276:immunology; D015800:Protozoan Proteins / Q000276:immunology* / Q000302:isolation & purification; D016052:Protozoan Vaccines / Q000008:administration & dosage; D011817:Rabbits; D051381:Rats; D014346:Trypanosoma brucei brucei / Q000276:immunology*; D014353:Trypanosomiasis, African / Q000276:immunology / Q000517:prevention & control*; D014404:Tubulin / Q000276:immunology* / Q000302:isolation & purification
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D007106:Immune Sera; D010455:Peptides; D015800:Protozoan Proteins; D016052:Protozoan Vaccines; D014404:Tubulin