The magnitude and breadth of T-cell responses against HCV are associated with the outcome of HCV infection. Parameters of HCV-specific T-cell responses that are frequently assessed in clinical immunological studies include proliferation of T cells in response to HCV antigens, frequency of HCV-specific T cells secreting cytokines, and changes in antigen specificity during the course of HCV infection. Common techniques for assessing these parameters such as (3)H-thymidine incorporation assay, cytokine ELISpot assay, and strategies for epitope mapping and identification of minimal optimal epitopes are outlined, and detailed protocols are described.
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http://dx.doi.org/10.1007/978-1-59745-394-3_30 | DOI Listing |
Viruses
October 2024
Centre de Recherche du Centre Hospitalier de l'Université de Montréal (CRCHUM), Tour Viger, Local R09.414, 900 rue St-Denis, Montréal, QC H2X 0A9, Canada.
Identification and isolation of antigen-specific T cells for downstream transcriptomic analysis is key for various immunological studies. Traditional methods using major histocompatibility complex (MHC) multimers are limited by the number of predefined immunodominant epitopes and MHC matching of the study subjects. Activation-induced markers (AIM) enable highly sensitive detection of rare antigen-specific T cells irrespective of the availability of MHC multimers.
View Article and Find Full Text PDFViruses
August 2024
Department of Biology and Biotechnology "Charles Darwin", Sapienza University of Rome, 00100 Rome, Italy.
Therapy against the Hepatitis C virus (HCV) has significantly improved with the introduction of direct-acting antiviral drugs (DAAs), achieving over 95% sustained virological response (SVR). Despite this, the development of an effective anti-HCV vaccine remains a critical challenge due to the low number of patients treated with DAAs and the occurrence of HCV reinfections in high-risk groups. Current vaccine strategies aim to stimulate either B-cell or T-cell responses.
View Article and Find Full Text PDFVaccine
September 2024
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:
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.
View Article and Find Full Text PDFFront Immunol
July 2024
Centre de Recherche du Centre Hospitalier de l'Université de Montréal (CRCHUM), Montréal, QC, Canada.
Introduction: Follicular helper T cells are essential for helping in the maturation of B cells and the production of neutralizing antibodies (NAbs) during primary viral infections. However, their role during recall responses is unclear. Here, we used hepatitis C virus (HCV) reinfection in humans as a model to study the recall collaborative interaction between circulating CD4 T follicular helper cells (cTfh) and memory B cells (MBCs) leading to the generation of NAbs.
View Article and Find Full Text PDFImmunol Cell Biol
August 2024
School of Biomedical Sciences, Faculty of Health and Medicine, UNSW Sydney, Sydney, NSW, Australia.
CD8 T cells recognizing their cognate antigen are typically recruited as a polyclonal population consisting of multiple clonotypes with varying T-cell receptor (TCR) affinity to the target peptide-major histocompatibility complex (pMHC) complex. Advances in single-cell sequencing have increased accessibility toward identifying TCRs with matched antigens. Here we present the discovery of a monoclonal CD8 T-cell population with specificity for a hepatitis C virus (HCV)-derived human leukocyte antigen (HLA) class I epitope (HLA-B*07:02 GPRLGVRAT) which was isolated directly ex vivo from an individual with an episode of acutely resolved HCV infection.
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