Hostname: page-component-cd9895bd7-fscjk Total loading time: 0 Render date: 2024-12-27T12:15:23.622Z Has data issue: false hasContentIssue false
  • English
  • Français

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

Published online by Cambridge University Press:  06 April 2009

L. Quijada ,
J. M. Requena ,
M. Soto  and
C. Alonso
L. Quijada
Affiliation:
Centra de Biología Molecular ‘Severo Ochoa’ (CSIC-UAM), Universidad Autónoma de Madrid, Cantoblanco, Madrid 28049, Spain
J. M. Requena
Affiliation:
Centra de Biología Molecular ‘Severo Ochoa’ (CSIC-UAM), Universidad Autónoma de Madrid, Cantoblanco, Madrid 28049, Spain
M. Soto
Affiliation:
Centra de Biología Molecular ‘Severo Ochoa’ (CSIC-UAM), Universidad Autónoma de Madrid, Cantoblanco, Madrid 28049, Spain
C. Alonso
Affiliation:
Centra de Biología Molecular ‘Severo Ochoa’ (CSIC-UAM), Universidad Autónoma de Madrid, Cantoblanco, Madrid 28049, Spain
Get access Rights & Permissions [Opens in a new window]

Summary

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.

Keywords

Leishmania infantumviscero-cutaneous leishmaniasishumoral responserecombinant proteinsamino acid sequence.
Type
Research Article
Information
Parasitology , Volume 112 , Issue 3 , March 1996 , pp. 277 - 284
Copyright
Copyright © Cambridge University Press 1996

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

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.CrossRefGoogle Scholar
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.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle Scholar
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.CrossRefGoogle Scholar
Devereux, J., Haeberli, P. & Smithies, O. (1984). A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Research 12, 387–95.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle Scholar
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.CrossRefGoogle ScholarPubMed
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. 49–78. Oxford: IRL Press.Google Scholar
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.CrossRefGoogle ScholarPubMed
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227, 680–5.CrossRefGoogle ScholarPubMed
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.Google ScholarPubMed
Lindquist, S. (1986). The heat shock response. Annual Reviews of Biochemistry 55, 1151–91.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle Scholar
Maresca, B. & Kobayashi, G. S. (1994). Hsp70 in parasites: as an inducible protective protein and as an antigen. Experientia 50, 10067–74.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle Scholar
Minowada, G. & Welch, W. J. (1995). Clinical implications of the stress response. Journal of Clinical Investigations 95, 3–12.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle Scholar
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.CrossRefGoogle Scholar
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, 299–308.CrossRefGoogle ScholarPubMed
Shinnick, T. M. (1991). Heat shock proteins as antigens of bacterial and parasitic pathogens. Current Topics in Microbiology and Immunology 167, 145–60.Google ScholarPubMed
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.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle Scholar
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.CrossRefGoogle ScholarPubMed
Winfield, J. B. & Jarjour, W. N. (1991). Stress proteins, autoimmunity, and autoimmune disease. Current Topics in Microbiology and Immunology 167, 161–89.Google ScholarPubMed
Young, D. B. (1992). Heat-shock proteins: immunity and autoimmunity. Current Opinion in Immunology 4, 396–400.CrossRefGoogle ScholarPubMed
Zhong, G. & Brunham, R. C. (1992). Antigenic analysis of the chlamydial 75-kilodalton protein. Infection and Immunity 60, 1221–34.CrossRefGoogle ScholarPubMed