Skip to main content Accessibility help

During canine viscero-cutaneous leishmaniasis the anti-Hsp70 antibodies are specifically elicited by the parasite protein*

  • L. Quijada (a1), J. M. Requena (a1), M. Soto (a1) and C. Alonso (a1)


A Leishmania infantum cDNA library was screened with sera from dogs with viscero-cutaneous leishmaniasis. Sequence analysis of a positive clone isolated from the library revealed that it coded for the carboxyl-terminal region of a member of the 70-kDa heat-shock protein family. The full-length sequence of the L. infantum hsp70 gene was determined after isolation of genomic clones. This protein shows a high degree of sequence conservation with the homologous protein from other organisms. To test its antigenicity a recombinant Hsp70 protein fused to the maltose-binding protein was produced in Escherichia coli using the expression vector pMAL-cRI. By FAST-ELISA assays it was observed that while the complete recombinant protein was recognized by 100% of the sera, the 20 carboxyl-terminal amino acids of the protein were only recognized by 30% of those sera. Thus, although a B-cell epitope must be present within the carboxyl terminal end of the protein other antigenic determinant(s) must reside out of this region. The analysis of the cross-reactivity with mouse Hsp70 by Western blotting strongly suggests that the anti-Hsp70 antibodies generated by infection with L. infantum are directed at specific determinants of the L. infantum Hsp70. Thus, our results indicate that anti-Hsp70 autoantibodies are not induced during Leishmania infection.



Hide All
Ardeshir, F., Flint, J. E., Richman, S. J., Reese, R. T. (1987). A 75 kd merozoite surface protein of Plasmodium falciparum which is related to the 70 kd heat-shock proteins. EMBO Journal 6, 493–9.
Behr, C., Sarthou, J.-L., Rogier, C., Trape, J.-F., Dat, M. H. Q., Michel, J.-C., Aribot, G., Dieye, A., Claverie, J.-M., Druihle, P. & Dubois, P. (1992). Antibodies and reactive T cells against the malaria heat-shock protein Pf72/hsp70–1 and derived peptides in individuals continuously exposed to Plasmodium falciparum. Journal of Immunology 149, 3321–30.
Bianco, A. E., Favaloro, J. M., Burkot, T. R., Culvenor, P. E., Brown, G. V., Anders, R. F., Coppel, R. L. & Kemp, D. J. (1986). A repetitive antigen of Plasmodium falciparum that is homologous to heat shock protein 70 of Drosophila melanogaster. Proceedings of the National Academy of Sciences, USA 83, 8713–17.
Bock, J. H. & Lancer, P. J. (1993). Sequence and genomic organization of the hsp70 genes of Leishmania amazonensis. Molecular and Biochemical Parasitology 62, 187–98.
Davenport, M. P., McKenzie, K. R., Basten, A. & Britton, W. J. (1992). The variable C-terminal region of the Mycobacterium leprae 70-kilodalton heat shock protein is the target for humoral immune response. Infection and Immunity 60, 1170–7.
De Andrade, C. R., Kirchhoff, L. V., Donelson, J. E. & Otsu, K. (1992). Recombinant Leishmania Hsp90 and Hsp70 are recognized by sera from visceral leishmaniasis patients but not Chagas' disease patients. Journal of Clinical Microbiology 30, 330–5.
Devereux, J., Haeberli, P. & Smithies, O. (1984). A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Research 12, 387–95.
Dworniczak, B. & Miraclt, M.-E. (1987). Structure and expression of a human gene coding for a 71 kd heat shock ‘cognate’ protein. Nucleic Acids Research 15, 5181–97.
Engman, D. M., Dragon, E. A. & Donelson, J. E. (1990). Human humoral immunity to hsp70 during Trypanosoma cruzi infection. Journal of Immunology 144, 3987–91.
Garsia, R. J., Hellqvist, L., Booth, R. J., Radford, A. J., Britton, W. J., Astbury, L., Trent, R. J. & Basten, A. (1989). Homology of the 70-kilodalton antigens from Mycobacterium leprae and Mycobacterium bovis with the Mycobacterium tuberculosis 71-kilodalton antigen and with the conserved heat shock protein 70 of eucaryotes. Infection and Immunity 57, 204–12.
Hedstrom, R., Culpepper, J., Harrison, R. A., Agabian, N. & Newport, G. (1987). A major immunogen in Schistosoma mansoni infections is homologous to the heat-shock protein hsp70. Journal of Experimental Medicine 165, 1430–5.
Hedstrom, R., Culpepper, J., Schinski, V., Agabian, N. & Newport, G. (1988). Schistosome heat-shock proteins are immunologically distinct host-like antigens. Molecular and Biochemical Parasitology 29, 275–82.
Huynh, T. V., Young, R. A. & Davis, R. W. (1985). Construction and screening cDNA libraries in λgt10 and λgt11. In DNA Cloning: A Practical Approach (ed. Glover, D. M.), pp. 4978. Oxford: IRL Press.
Kumar, N., Zhao, Y., Graves, P., Folgar, J. P., Maloy, L. & Zheng, H. (1990). Human immune response directed against Plasmodium falciparum heat shock-related proteins. Infection and Immunity 58, 1408–14.
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227, 680–5.
Lathigra, R. B., Butcher, P. D., Garbe, T. R. & Young, D. B. (1991). Heat shock proteins as virulence factors of pathogens. Current Topics in Microbiology and Immunology 167, 125–43.
Lindquist, S. (1986). The heat shock response. Annual Reviews of Biochemistry 55, 1151–91.
MacFarlane, J., Blaxter, M. L., Bishop, R. P. & Kelly, J. M. (1990). Identification and characterisation of a Leishmania donovani antigen belonging to the 70-kDa heat-shock protein family. European Journal of Biochemistry 190, 377–84.
Maresca, B. & Kobayashi, G. S. (1994). Hsp70 in parasites: as an inducible protective protein and as an antigen. Experientia 50, 10067–74.
Mattei, D., Scherf, A., Bensaude, O. & Da Silva, L. P. (1989). A heat shock-like protein from the human malaria parasite Plasmodium falciparum induces autoantibodies. European Journal of Immunology 19, 1823–8.
McKenzie, K. R., Adams, E., Britton, W. J., Garsia, R. J. & Basten, A. (1991). Sequence and immunogenicity of the 70-kDa heat shock protein of Mycobacterium leprae. Journal of Immunology 147, 312–19.
Minota, S., Cameron, B., Welch, W. J. & Winfield, J. B. (1988). Autoantibodies to the constitutive 73-kD member of the hsp70 family of heat shock proteins in systemic lupus erythematosus. Journal of Experimental Medicine 168, 1475–80.
Minowada, G. & Welch, W. J. (1995). Clinical implications of the stress response. Journal of Clinical Investigations 95, 312.
Nafziger, D. A., Recinos, R. F., Hunter, C. A. & Donelson, J. E. (1991). Patients infected with Leishmania donovani chagasi can have antibodies that recognize heat shock and acidic ribosomal proteins of Trypanosoma cruzi. Molecular and Biochemical Parasitology 49, 325–8.
Requena, J. M., Lopez, M. C., Jimenez-Ruiz, A., Morales, G. & Alonso, C. (1989). Complete nucleotide sequence of the hsp70 gene of T. cruzi. Nucleic Acids Research 17, 797.
Requena, J. M., Soto, M., Guzman, F., Maekel, A., Noya, O., Patarroyo, M. E. & Alonso, C. (1993). Mapping of antigenic determinants of the T. cruzi hsp70 in chagasic and healthy individuals. Molecular Immunology 30, 1115–21.
Rothstein, N. M., Higashi, G., Yates, J. & Rajan, T. V. (1989). Onchocerca volvulus heat shock protein 70 is a major immunogen in amicrofilaremic individuals from a filariasis-endemic area. Molecular and Biochemical Parasitology 33, 229–36.
Selkirk, M. E., Denham, D. A., Partono, F. & Maizels, R. M. (1989). Heat shock cognate 70 is a prominent immunogen in Brugian filariasis. Journal of Immunology 143, 299308.
Shinnick, T. M. (1991). Heat shock proteins as antigens of bacterial and parasitic pathogens. Current Topics in Microbiology and Immunology 167, 145–60.
Srivastava, P. K., Udono, H., Blachere, N. E. & Li, Z. (1994). Heat shock proteins transfer peptides during antigen processing and CTL priming. Immunogenetics 39, 93–8.
Velazquez, J. M., Sonoda, S., Bugaisky, G. E. & Lindquist, S. (1983). Are Hs proteins present in cells that have not been heat shocked? Journal of Cell Biology 96, 286–90.
Wallace, G. R., Ball, A. E., MacFarlane, J., El Safi, S. H., Miles, M. A. & Kelly, J. M. (1992). Mapping of a visceral leishmaniasis-specific immunodominant B-cell epitope of Leishmania donovani hsp70. Infection and Immunity 60, 2688–93.
Winfield, J. B. & Jarjour, W. N. (1991). Stress proteins, autoimmunity, and autoimmune disease. Current Topics in Microbiology and Immunology 167, 161–89.
Young, D. B. (1992). Heat-shock proteins: immunity and autoimmunity. Current Opinion in Immunology 4, 396400.
Zhong, G. & Brunham, R. C. (1992). Antigenic analysis of the chlamydial 75-kilodalton protein. Infection and Immunity 60, 1221–34.



Altmetric attention score

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed