دورية أكاديمية
An in vitro CD8 T-cell priming assay enables epitope selection for hepatitis C virus vaccines.
| العنوان: | An in vitro CD8 T-cell priming assay enables epitope selection for hepatitis C virus vaccines. |
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| المؤلفون: | Koutsoumpli G; Department of Medical Microbiology and Infection Prevention, University Medical Center Groningen, University of Groningen, PO Box 30 001, HPC EB88, 9700RB Groningen, the Netherlands., Stasiukonyte N; Department of Medical Microbiology and Infection Prevention, University Medical Center Groningen, University of Groningen, PO Box 30 001, HPC EB88, 9700RB Groningen, the Netherlands., Hoogeboom BN; Department of Medical Microbiology and Infection Prevention, University Medical Center Groningen, University of Groningen, PO Box 30 001, HPC EB88, 9700RB Groningen, the Netherlands., Daemen T; Department of Medical Microbiology and Infection Prevention, University Medical Center Groningen, University of Groningen, PO Box 30 001, HPC EB88, 9700RB Groningen, the Netherlands. Electronic address: c.a.h.h.daemen@umcg.nl. |
| المصدر: | Vaccine [Vaccine] 2024 Sep 17; Vol. 42 (22), pp. 126032. Date of Electronic Publication: 2024 Jul 03. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Elsevier Science Country of Publication: Netherlands NLM ID: 8406899 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1873-2518 (Electronic) Linking ISSN: 0264410X NLM ISO Abbreviation: Vaccine Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Amsterdam, The Netherlands : Elsevier Science Original Publication: [Guildford, Surrey, UK] : Butterworths, [c1983- |
| مواضيع طبية MeSH: | CD8-Positive T-Lymphocytes*/immunology , Epitopes, T-Lymphocyte*/immunology , Hepacivirus*/immunology , Hepacivirus*/genetics , Viral Hepatitis Vaccines*/immunology , Viral Nonstructural Proteins*/immunology, Humans ; Hepatitis C/immunology ; Hepatitis C/prevention & control ; Enzyme-Linked Immunospot Assay/methods ; HLA-A2 Antigen/immunology ; Interferon-gamma/immunology ; Interferon-gamma/metabolism ; Viral Proteases ; Serine Endopeptidases ; Nucleoside-Triphosphatase ; DEAD-box RNA Helicases |
| مستخلص: | For the rational design of epitope-specific vaccines, identifying epitopes that can be processed and presented is essential. As algorithm-based epitope prediction is frequently discordant with actually recognized CD8 + T-cell epitopes, we developed an in vitro CD8 T-cell priming protocol to enable the identification of truly and functionally expressed HLA class I epitopes. The assay was established and validated to identify epitopes presented by hepatitis C virus (HCV)-infected cells. In vitro priming of naïve CD8 T cells was achieved by culturing unfractionated PBMCs in the presence of a specific cocktail of growth factors and cytokines, and next exposing the cells to hepatic cells expressing the NS3 protein of HCV. After a 10-day co-culture, HCV-specific T-cell responses were identified based on IFN-γ ELISpot analysis. For this, the T cells were restimulated with long synthetic peptides (SLPs) spanning the whole NS3 protein sequence allowing the identification of HCV-specificity. We demonstrated that this protocol resulted in the in vitro priming of naïve precursors to antigen-experienced T-cells specific for 11 out of 98 SLPs tested. These 11 SLPs contain 12 different HLA-A*02:01-restricted epitopes, as predicted by a combination of three epitope prediction algorithms. Furthermore, we identified responses against 3 peptides that were not predicted to contain any immunogenic HLA class I epitopes, yet showed HCV-specific responses in vitro. Separation of CD8 + and CD8 - T cells from PBMCs primed in vitro showed responses only upon restimulation with short peptides. We established an in vitro method that enables the identification of HLA class I epitopes resulting from cross-presented antigens and that can cross-prime T cells and allows the effective selection of functional immunogenic epitopes, but also less immunogenic ones, for the design of tailored therapeutic vaccines against persistent viral infections and tumor antigens. Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.) |
| فهرسة مساهمة: | Keywords: CD8(+) T-cell priming assay; Cancer vaccine; Epitope identification; hepatitis C virus |
| المشرفين على المادة: | 0 (Epitopes, T-Lymphocyte) 0 (Viral Hepatitis Vaccines) 0 (Viral Nonstructural Proteins) 0 (NS3 protein, hepatitis C virus) 0 (HLA-A2 Antigen) 82115-62-6 (Interferon-gamma) EC 3.4.- (Viral Proteases) EC 3.4.21.- (Serine Endopeptidases) EC 3.6.1.15 (Nucleoside-Triphosphatase) EC 3.6.4.13 (DEAD-box RNA Helicases) |
| تواريخ الأحداث: | Date Created: 20240704 Date Completed: 20240902 Latest Revision: 20240902 |
| رمز التحديث: | 20240903 |
| DOI: | 10.1016/j.vaccine.2024.05.080 |
| PMID: | 38964950 |
| قاعدة البيانات: | MEDLINE |
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Full Text Finder | تدمد: | 1873-2518 |
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| DOI: | 10.1016/j.vaccine.2024.05.080 |