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

Parasitology. 1996 Mar;112 ( Pt 3):277-84. doi: 10.1017/s0031182000065793.

Abstract

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.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Protozoan / biosynthesis*
  • Antibody Formation
  • Antibody Specificity
  • Autoantibodies / biosynthesis*
  • Blotting, Western
  • Cloning, Molecular
  • Conserved Sequence
  • DNA, Complementary
  • Dog Diseases*
  • Dogs
  • Gene Library
  • Genes, Protozoan
  • HSP70 Heat-Shock Proteins / biosynthesis
  • HSP70 Heat-Shock Proteins / genetics
  • HSP70 Heat-Shock Proteins / immunology*
  • Leishmania infantum* / genetics
  • Leishmania infantum* / immunology
  • Leishmaniasis, Cutaneous / immunology
  • Leishmaniasis, Cutaneous / veterinary*
  • Leishmaniasis, Visceral / immunology
  • Leishmaniasis, Visceral / veterinary*
  • Mice
  • Molecular Sequence Data
  • Protozoan Proteins / biosynthesis
  • Protozoan Proteins / genetics
  • Protozoan Proteins / immunology*
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / immunology
  • Sequence Homology, Amino Acid

Substances

  • Antibodies, Protozoan
  • Autoantibodies
  • DNA, Complementary
  • HSP70 Heat-Shock Proteins
  • Protozoan Proteins
  • Recombinant Proteins

Associated data

  • GENBANK/X85798