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).
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.
Delgado
Gabriela
G
Fundación Instituto de Inmunología de Colombia (FIDIC), Departamento de Farmacia, Facultad de Ciencios, Universidad Nacional de Colombia, Universidad Autonoma de Madrid, Spain.
Parra-López
Carlos A
CA
Vargas
Luis Eduardo
LE
Hoya
Rubén
R
Estupiñán
Mónica
M
Guzmán
Fanny
F
Torres
Angela
A
Alonso
Carlos
C
Velez
Iván Dario
ID
Spinel
Clara
C
Patarroyo
Manuel E
ME
D000595:Amino Acid Sequence; D000818:Animals; D017951:Antigen Presentation; D000953:Antigens, Protozoan / Q000235:genetics; D001667:Binding, Competitive; D016022:Case-Control Studies; D002460:Cell Line; D016207:Cytokines / Q000096:biosynthesis; D003713:Dendritic Cells / Q000276:immunology; D018604:Epitope Mapping; D000939:Epitopes / Q000235:genetics; D005260:Female; D006684:HLA-DR Antigens / Q000378:metabolism; D059811:HLA-DRB1 Chains; D006801:Humans; D007111:Immunity, Cellular; D066298:In Vitro Techniques; D018334:Leishmania guyanensis / Q000235:genetics / Q000276:immunology*; D007897:Leishmaniasis, Mucocutaneous / Q000276:immunology*; D008213:Lymphocyte Activation; D008297:Male; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D015800:Protozoan Proteins / Q000235:genetics / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology
D000953:Antigens, Protozoan; D016207:Cytokines; D000939:Epitopes; D006684:HLA-DR Antigens; D059811:HLA-DRB1 Chains; C553505:HLA-DRB1*04:01 antigen; D008565:Membrane Proteins; D015800:Protozoan Proteins