PMID- 32176691 OWN - NLM STAT- MEDLINE DCOM- 20200512 LR - 20200518 IS - 1935-2735 (Electronic) IS - 1935-2727 (Print) IS - 1935-2727 (Linking) VI - 14 IP - 3 DP - 2020 Mar TI - Design of multi-epitope peptides containing HLA class-I and class-II-restricted epitopes derived from immunogenic Leishmania proteins, and evaluation of CD4+ and CD8+ T cell responses induced in cured cutaneous leishmaniasis subjects. PG - e0008093 LID - 10.1371/journal.pntd.0008093 [doi] LID - e0008093 AB - Human leishmaniasis is a public health problem worldwide for which the development of a vaccine remains a challenge. T cell-mediated immune responses are crucial for protection. Peptide vaccines based on the identification of immunodominant T cell epitopes able to induce T cell specific immune responses constitute a promising strategy. Here, we report the identification of human leukocyte antigen class-I (HLA-I) and -II (HLA-II)-restricted multi-epitope peptides from Leishmania proteins that we have previously described as vaccine candidates. Promastigote Surface Antigen (PSA), LmlRAB (L. major large RAB GTPase) and Histone (H2B) were screened, in silico, for T cell epitopes. 6 HLA-I and 5 HLA-II-restricted multi-epitope peptides, able to bind to the most frequent HLA molecules, were designed and used as pools to stimulate PBMCs from individuals with healed cutaneous leishmaniasis. IFN-gamma, IL-10, TNF-alpha and granzyme B (GrB) production was evaluated by ELISA/CBA. The frequency of IFN-gamma-producing T cells was quantified by ELISpot. T cells secreting cytokines and memory T cells were analyzed by flow cytometry. 16 of 25 peptide pools containing HLA-I, HLA-II or HLA-I and -II peptides were able to induce specific and significant IFN-gamma levels. No IL-10 was detected. 6 peptide pools were selected among those inducing the highest IFN-gamma levels for further characterization. 3/6 pools were able to induce a significant increase of the percentages of CD4+IFN-gamma+, CD8+IFN-gamma+ and CD4+GrB+ T cells. The same pools also induced a significant increase of the percentages of bifunctional IFN-gamma+/TNF-alpha+CD4+ and/or central memory T cells. We identified highly promiscuous HLA-I and -II restricted epitope combinations from H2B, PSA and LmlRAB proteins that stimulate both CD4+ and CD8+ T cell responses in recovered individuals. These multi-epitope peptides could be used as potential components of a polytope vaccine for human leishmaniasis. FAU - Hamrouni, Sarra AU - Hamrouni S AD - Laboratoire de Parasitologie Medicale, Biotechnologie et Biomolecules, Institut Pasteur de Tunis, Tunis, Tunisie. AD - Faculte des Sciences de Bizerte, Universite de Carthage, Tunis, Tunisie. AD - UMR INTERTRYP, Universite de Montpellier, IRD, CIRAD, Montpellier, France. FAU - Bras-Goncalves, Rachel AU - Bras-Goncalves R AUID- ORCID: 0000-0002-3089-4530 AD - UMR INTERTRYP, Universite de Montpellier, IRD, CIRAD, Montpellier, France. FAU - Kidar, Abdelhamid AU - Kidar A AD - Hopital Regional de Gafsa, Gafsa, Tunisia. FAU - Aoun, Karim AU - Aoun K AD - Laboratoire de Parasitologie Medicale, Biotechnologie et Biomolecules, Institut Pasteur de Tunis, Tunis, Tunisie. FAU - Chamakh-Ayari, Rym AU - Chamakh-Ayari R AD - Laboratoire de Parasitologie Medicale, Biotechnologie et Biomolecules, Institut Pasteur de Tunis, Tunis, Tunisie. AD - Faculte des Sciences de Bizerte, Universite de Carthage, Tunis, Tunisie. FAU - Petitdidier, Elodie AU - Petitdidier E AUID- ORCID: 0000-0002-8554-8424 AD - UMR INTERTRYP, Universite de Montpellier, IRD, CIRAD, Montpellier, France. FAU - Messaoudi, Yasmine AU - Messaoudi Y AUID- ORCID: 0000-0003-0033-2729 AD - Laboratoire de Parasitologie Medicale, Biotechnologie et Biomolecules, Institut Pasteur de Tunis, Tunis, Tunisie. AD - Faculte des Sciences de Bizerte, Universite de Carthage, Tunis, Tunisie. AD - UMR INTERTRYP, Universite de Montpellier, IRD, CIRAD, Montpellier, France. FAU - Pagniez, Julie AU - Pagniez J AD - UMR INTERTRYP, Universite de Montpellier, IRD, CIRAD, Montpellier, France. FAU - Lemesre, Jean-Loup AU - Lemesre JL AUID- ORCID: 0000-0002-7406-8820 AD - UMR INTERTRYP, Universite de Montpellier, IRD, CIRAD, Montpellier, France. FAU - Meddeb-Garnaoui, Amel AU - Meddeb-Garnaoui A AUID- ORCID: 0000-0002-1581-2779 AD - Laboratoire de Parasitologie Medicale, Biotechnologie et Biomolecules, Institut Pasteur de Tunis, Tunis, Tunisie. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20200316 PL - United States TA - PLoS Negl Trop Dis JT - PLoS neglected tropical diseases JID - 101291488 RN - 0 (Antigens, Protozoan) RN - 0 (Epitopes, T-Lymphocyte) RN - 0 (Histocompatibility Antigens Class I) RN - 0 (Histocompatibility Antigens Class II) RN - 0 (IL10 protein, human) RN - 0 (Recombinant Fusion Proteins) RN - 0 (Tumor Necrosis Factor-alpha) RN - 130068-27-8 (Interleukin-10) RN - 82115-62-6 (Interferon-gamma) RN - EC 3.4.21.- (Granzymes) SB - IM MH - Adult MH - Antigens, Protozoan/genetics/*immunology MH - CD4-Positive T-Lymphocytes/*immunology MH - CD8-Positive T-Lymphocytes/*immunology MH - Enzyme-Linked Immunosorbent Assay MH - Epitopes, T-Lymphocyte/genetics/*immunology MH - Female MH - Flow Cytometry MH - Granzymes/analysis MH - Histocompatibility Antigens Class I/metabolism MH - Histocompatibility Antigens Class II/metabolism MH - Humans MH - Interferon-gamma/analysis MH - Interleukin-10/analysis MH - Leishmania/*immunology MH - Leishmaniasis, Cutaneous/*immunology MH - Male MH - Middle Aged MH - Protein Binding MH - Recombinant Fusion Proteins/genetics/*immunology/metabolism MH - Tumor Necrosis Factor-alpha/analysis MH - Volunteers MH - Young Adult PMC - PMC7098648 COIS- The authors have declared that no competing interests exist. EDAT- 2020/03/17 06:00 MHDA- 2020/05/19 06:00 PMCR- 2020/03/16 CRDT- 2020/03/17 06:00 PHST- 2019/07/09 00:00 [received] PHST- 2020/01/27 00:00 [accepted] PHST- 2020/03/26 00:00 [revised] PHST- 2020/03/17 06:00 [pubmed] PHST- 2020/05/19 06:00 [medline] PHST- 2020/03/17 06:00 [entrez] PHST- 2020/03/16 00:00 [pmc-release] AID - PNTD-D-19-01171 [pii] AID - 10.1371/journal.pntd.0008093 [doi] PST - epublish SO - PLoS Negl Trop Dis. 2020 Mar 16;14(3):e0008093. doi: 10.1371/journal.pntd.0008093. eCollection 2020 Mar.