Characterizing cellular immune response to kinetoplastid membrane protein-11 (KMP-11) during Leishmania (Viannia) panamensis infection using dendritic cells (DCs) as antigen presenting cells (APCs)

Parasite Immunol. 2003 Apr;25(4):199-209. doi: 10.1046/j.1365-3024.2003.00626.x.

Abstract

In vitro peptide binding assays and DCs pulsed with recombinant KMP-11 (rKMP-11) plus six 20-mer overlapping peptides covering the entire protein of Leishmania (Viannia) panamensis (L(V)p) promastigotes were used to identify T-cell epitopes in this protein. Such in vitro binding assays, using HLA DRB1* 0101, -0401, -0701 and -1101 alleles, demonstrated that two peptide sequences (DEEFNKKMQEQNAKFFADKP and FKHKFAELLEQQKAAQYPSK) exhibited high HLA DRB1* 0401 allele binding capacity. rKMP-11 specific T-cell proliferation and cytokine production, derived from 13 volunteers exposed to the parasite, suggested that using autologous DCs as APCs becomes advantageous in uncovering T-cell epitopes promoting proliferation and differences in IFN-gamma and IL-4 production in T-cells from volunteers with ACTIVE and CURED undetectable disease when other APCs were used. The two peptides which bound in vitro to the HLA DRB1* 0401 allele were immunogenic in HLA DRB1* 04 volunteers, thus validating the use of in vitro binding assays for predicting epitopes in this protein. The experimental approach used here may prove useful for characterizing T-cell epitopes in a protein useful in designing peptide-based vaccine candidates for Leishmania and other intracellular pathogens.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antigen Presentation
  • Antigens, Protozoan / genetics
  • Binding, Competitive
  • Case-Control Studies
  • Cell Line
  • Cytokines / biosynthesis
  • Dendritic Cells / immunology
  • Epitope Mapping
  • Epitopes / genetics
  • Female
  • HLA-DR Antigens / metabolism
  • HLA-DRB1 Chains
  • Humans
  • Immunity, Cellular*
  • In Vitro Techniques
  • Leishmania guyanensis / genetics
  • Leishmania guyanensis / immunology*
  • Leishmaniasis, Mucocutaneous / immunology*
  • Lymphocyte Activation
  • Male
  • Membrane Proteins / genetics
  • Membrane Proteins / immunology*
  • Molecular Sequence Data
  • Protozoan Proteins / genetics
  • Protozoan Proteins / immunology*
  • T-Lymphocytes / immunology

Substances

  • Antigens, Protozoan
  • Cytokines
  • Epitopes
  • HLA-DR Antigens
  • HLA-DRB1 Chains
  • HLA-DRB1*04:01 antigen
  • Membrane Proteins
  • Protozoan Proteins