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Human leukocyte antigen (HLA) class I restricted epitope discovery in yellow fewer and dengue viruses: importance of HLA binding strength.

Lund O, Nascimento EJ, Maciel M, Nielsen M, Larsen MV, Lundegaard C, Harndahl M, Lamberth K, Buus S, Salmon J, August TJ, Marques ET - PLoS ONE (2011)

Bottom Line: The selected predicted epitopes were synthesized and approximately 75% were found to bind the predicted restricting HLA molecule with an affinity, K(D), stronger than 500 nM.The immunogenic peptides bound HLA significantly stronger than the non-immunogenic peptides.The results indicate the importance of the strength of HLA binding in shaping the immune response.

View Article: PubMed Central - PubMed

Affiliation: Department of Systems Biology, Center for Biological Sequence Analysis, Technical University of Denmark, Lyngby, Denmark. lund@cbs.dtu.dk

ABSTRACT
Epitopes from all available full-length sequences of yellow fever virus (YFV) and dengue fever virus (DENV) restricted by Human Leukocyte Antigen class I (HLA-I) alleles covering 12 HLA-I supertypes were predicted using the NetCTL algorithm. A subset of 179 predicted YFV and 158 predicted DENV epitopes were selected using the EpiSelect algorithm to allow for optimal coverage of viral strains. The selected predicted epitopes were synthesized and approximately 75% were found to bind the predicted restricting HLA molecule with an affinity, K(D), stronger than 500 nM. The immunogenicity of 25 HLA-A*02:01, 28 HLA-A*24:02 and 28 HLA-B*07:02 binding peptides was tested in three HLA-transgenic mice models and led to the identification of 17 HLA-A*02:01, 4 HLA-A*2402 and 4 HLA-B*07:02 immunogenic peptides. The immunogenic peptides bound HLA significantly stronger than the non-immunogenic peptides. All except one of the immunogenic peptides had K(D) below 100 nM and the peptides with K(D) below 5 nM were more likely to be immunogenic. In addition, all the immunogenic peptides that were identified as having a high functional avidity had K(D) below 20 nM. A*02:01 transgenic mice were also inoculated twice with the 17DD YFV vaccine strain. Three of the YFV A*02:01 restricted peptides activated T-cells from the infected mice in vitro. All three peptides that elicited responses had an HLA binding affinity of 2 nM or less. The results indicate the importance of the strength of HLA binding in shaping the immune response.

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ELISPOT results in 9 mice vaccinated with either A*02:01 dengue peptide pool or YFV 17DD vaccine in A*02:01 mice.Peptides positive at concentration 0.1 µg/mL and greater are shown. Results are depicted as percentage of spot-forming cells/million cells of each peptide at 10 µg/mL.
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pone-0026494-g002: ELISPOT results in 9 mice vaccinated with either A*02:01 dengue peptide pool or YFV 17DD vaccine in A*02:01 mice.Peptides positive at concentration 0.1 µg/mL and greater are shown. Results are depicted as percentage of spot-forming cells/million cells of each peptide at 10 µg/mL.

Mentions: When the peptides giving significant responses were tested in vitro at different concentrations, five peptides (two DENV and three YFV) that induced responses in A*02:01 mice showed high functional avidity achieving approximately half the maximal of the T-cell responses at peptide concentrations as low as 0.1 µg/mL (Figure 2). The A*02:01 response was specific, since no peptide from the B*07:02 set (used as negative controls) induced any response in A*02:01 mice (data not shown). Three of the A*24:03 immunogenic peptides elicited high affinity responses (Figure 3). Note that all the peptides that were found to be immunogenic in transgenic B*07:02 mice only activated CTLs at the highest concentration used, 10 µg/ml. Since we have only made graphs of peptides that elicited a response at 0.1 µg/ml, they were not included in any figure.


Human leukocyte antigen (HLA) class I restricted epitope discovery in yellow fewer and dengue viruses: importance of HLA binding strength.

Lund O, Nascimento EJ, Maciel M, Nielsen M, Larsen MV, Lundegaard C, Harndahl M, Lamberth K, Buus S, Salmon J, August TJ, Marques ET - PLoS ONE (2011)

ELISPOT results in 9 mice vaccinated with either A*02:01 dengue peptide pool or YFV 17DD vaccine in A*02:01 mice.Peptides positive at concentration 0.1 µg/mL and greater are shown. Results are depicted as percentage of spot-forming cells/million cells of each peptide at 10 µg/mL.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3198402&req=5

pone-0026494-g002: ELISPOT results in 9 mice vaccinated with either A*02:01 dengue peptide pool or YFV 17DD vaccine in A*02:01 mice.Peptides positive at concentration 0.1 µg/mL and greater are shown. Results are depicted as percentage of spot-forming cells/million cells of each peptide at 10 µg/mL.
Mentions: When the peptides giving significant responses were tested in vitro at different concentrations, five peptides (two DENV and three YFV) that induced responses in A*02:01 mice showed high functional avidity achieving approximately half the maximal of the T-cell responses at peptide concentrations as low as 0.1 µg/mL (Figure 2). The A*02:01 response was specific, since no peptide from the B*07:02 set (used as negative controls) induced any response in A*02:01 mice (data not shown). Three of the A*24:03 immunogenic peptides elicited high affinity responses (Figure 3). Note that all the peptides that were found to be immunogenic in transgenic B*07:02 mice only activated CTLs at the highest concentration used, 10 µg/ml. Since we have only made graphs of peptides that elicited a response at 0.1 µg/ml, they were not included in any figure.

Bottom Line: The selected predicted epitopes were synthesized and approximately 75% were found to bind the predicted restricting HLA molecule with an affinity, K(D), stronger than 500 nM.The immunogenic peptides bound HLA significantly stronger than the non-immunogenic peptides.The results indicate the importance of the strength of HLA binding in shaping the immune response.

View Article: PubMed Central - PubMed

Affiliation: Department of Systems Biology, Center for Biological Sequence Analysis, Technical University of Denmark, Lyngby, Denmark. lund@cbs.dtu.dk

ABSTRACT
Epitopes from all available full-length sequences of yellow fever virus (YFV) and dengue fever virus (DENV) restricted by Human Leukocyte Antigen class I (HLA-I) alleles covering 12 HLA-I supertypes were predicted using the NetCTL algorithm. A subset of 179 predicted YFV and 158 predicted DENV epitopes were selected using the EpiSelect algorithm to allow for optimal coverage of viral strains. The selected predicted epitopes were synthesized and approximately 75% were found to bind the predicted restricting HLA molecule with an affinity, K(D), stronger than 500 nM. The immunogenicity of 25 HLA-A*02:01, 28 HLA-A*24:02 and 28 HLA-B*07:02 binding peptides was tested in three HLA-transgenic mice models and led to the identification of 17 HLA-A*02:01, 4 HLA-A*2402 and 4 HLA-B*07:02 immunogenic peptides. The immunogenic peptides bound HLA significantly stronger than the non-immunogenic peptides. All except one of the immunogenic peptides had K(D) below 100 nM and the peptides with K(D) below 5 nM were more likely to be immunogenic. In addition, all the immunogenic peptides that were identified as having a high functional avidity had K(D) below 20 nM. A*02:01 transgenic mice were also inoculated twice with the 17DD YFV vaccine strain. Three of the YFV A*02:01 restricted peptides activated T-cells from the infected mice in vitro. All three peptides that elicited responses had an HLA binding affinity of 2 nM or less. The results indicate the importance of the strength of HLA binding in shaping the immune response.

Show MeSH
Related in: MedlinePlus