Trypanosoma cruzi flagellar repetitive antigen expression by recombinant baculovirus: towards an improved diagnostic reagent for Chagas' disease.
We constructed a recombinant baculovirus that expressed part of a Trypanosoma cruzi flagellar repetitive antigen (FRA). Both cell- associated and secreted forms of recombinant FRA were detected in cultures of virus-infected Spodoptera frugiperda (Sf9) cells. These forms show a complex pattern after polyacrylamide gel electrophoresis and Western blot analysis using either an anti-FRA rabbit serum or human Chagasic sera. Competitive Western-blot experiments revealed that all bands react with the same antibodies as a bacterially-derived FRA. Polymerase chain reaction and Southern blots of the recombinant viral DNA also showed a complex pattern, suggesting the presence of more than one repeat unit in the viral genome. When tested against a panel of human sera from an endemic area for Chagas' disease, FRA recombinant-Sf9 culture supernatant showed the same reactivity as purified FRA produced in bacteria.
Department of Biochemistry and Molecular Biology, Inst. Oswaldo Cruz, Rio de Janeiro, Brazil.
C N
CN
dos Santos
M A
MA
Krieger
E
E
Almeida
J J
JJ
Lafaille
S
S
Goldenberg
R
R
Galler
D000818:Animals; D000953:Antigens, Protozoan / Q000276:immunology*; D016367:Baculoviridae / Q000276:immunology*; D001709:Biotechnology; D002478:Cells, Cultured; D014355:Chagas Disease / Q000175:diagnosis*; D004279:DNA, Viral / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D005407:Flagella / Q000276:immunology*; D007118:Immunoassay; D007313:Insecta / Q000166:cytology; D016133:Polymerase Chain Reaction; D014349:Trypanosoma cruzi; D014764:Viral Proteins / Q000032:analysis
D000953:Antigens, Protozoan; D004279:DNA, Viral; D014764:Viral Proteins