Research Paper | Published:

Trypanosoma Cruzi Flagellar Repetitive Antigen Expression by Recombinant Baculovirus: Towards an Improved Diagnostics Reagent for Chagas' Disease

Bio/Technologyvolume 10pages14741477 (1992) | Download Citation



We constructed a recombinant baculovirus that expressed part of a Trypanosorna 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.

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  1. 1

    Chagas, C. 1909. Nova tripanosomiase humana. Estudos sobre a morfolojia e o cyclo evolutivo do Schyzotrypanum cruzi n.gen., agente etiolojico de nowa morbidade do homem. Mem. Inst. Oswaldo Cruz 1: 159–218.

  2. 2

    Topical Diseases. Progress in Research, 1989–1990. Tenth Programme Report of the UNDP/World Bank/WHO Special Programme for Research and Training in Tropical Diseases (TDR). WHO, Geneva 1991.

  3. 3

    Schmunis, G.A. 1985. Chagas' disease in blood transfusion, p. 127. In: Infection, Immunity and Blood Transfusion. Alan R. Liss, Inc., NY.

  4. 4

    Moncayo, A. and Luquetti, A.O. 1990. Multicentre double blind study for evaluation of Trypanosoma cruzi defined antigens as diagnostic reagents. Mem. Inst. Oswaldo Cruz 85: 489–495.

  5. 5

    Lafaille, J.J., Linss, J., Krieger, M.A., Souto-Padron, T., de Souza, W., and Goldenberg, S. . 1989. Structure and expression of two Trypanosoma cruzi genes encoding antigenic proteins bearing repetitve epitopes. Mol. Biochem. Parasitol. 35: 127–136.

  6. 6

    Almeida, E., Krieger, M.A., Carvalho, M.R., Oelemann, W. and Goldenberg, S. 1990. Use of recombinant antigens for the diagnosis of Chagas' disease and blood-bank screening. Mem. Inst. Oswaldo Cruz 85: 513–517.

  7. 7

    Krieger, M.A., Almeida, E.C., Oelemann, W., Lafaille, J.J., Pereira, J.B., Krieger, H., Carvalho, M.R. and Goldenberg, S. 1992. Use of recombinant antigens for the accurate immunodiagnosis of Chagas' disease. Am. J. Trop. Med. Hyg. 46: 427–434.

  8. 8

    Matsuura, Y., Possee, R.D., Overton, H.A. and Bishop, D.H. 1987. Baculovirus expression vectors: The requirements for high level expression of proteins, including glycoproteins. J. Gen. Virol. 68: 1233–1250.

  9. 9

    Sanger, F., Nicklen, S. and Coulson, A.R. 1977. DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. USA 74: 5463–5467.

  10. 10

    de la Cruz, V.F., Lal, A.A. and McCutchan, T.F. 1987. Sequence variation in putative functional domains of the circumsporozoite protein of Plasmodium falciparum. J. Biol. Chem. 262: 11935–11939.

  11. 11

    Summers, M.D. and Smith, G.E. 1987. A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures. Texas Agricultural Experiment Station. Bulletin No 1555.

  12. 12

    Rice, C.M., Franke, C.A., Strauss, H.J. and Hruby, D.E. 1985. Expression of Sindbis virus structural proteins via recombinant vaccinia virus: synthesis, processing and incorporation into mature Sindbis virions. J. Virol. 56: 227–239.

  13. 13

    Maniatis, T., Fritsch, E.F. and Sambrook, J. 1982. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, New York.

  14. 14

    Saiki, R.K., Scharf, S., Falooma, F., Mullins, B.K., Horn, G., Erlich, H. and Arnhein, N. 1985. Enzymatic amplification of β-globin genomic sequences and restriction site analysis for diagnosis of sickle cell amemia. Science 230: 1350–1354.

  15. 15

    Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.

  16. 16

    Towbin, H., Staehelin, T. and Gordon, J. 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci. USA 76: 4350–4354.

  17. 17

    Krieger, M.A., Salles, J.M., Almeida, E., Linss, J., Bonaldo, M.C. and Goldenberg, S. 1990. Expression and polymorphism of a trypanosoma antigen encoding a cytoplasmic repetitive antigen. Exp. Parasitol. 70: 247–254.

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  1. Claudia N. Duarte dos Santos: Corresponding author.


  1. Department of Biochemistry and Molecular Biology, Inst. Oswaldo Cruz, Av. Brasil 4365, Rio de Janeiro, Brazil, 21045

    • Claudia N. Duarte dos Santos
    • , Marco A. Krieger
    • , Elza Almeida
    • , Juan J. Lafaille
    • , Samuel Goldenberg
    •  & Ricardo Galler


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