Enhanced Detection of Antigen-Specific CD4+ T Cells Using Altered Peptide Flanking Residue Peptide–MHC Class II Multimers

Holland, Christopher; Dolton, Garry; Scurr, Martin; Ladell, Kristin; Schauenburg, Andrea J. A.; Miners, Kelly L.; Madura, Florian; Sewell, Andrew K.; Price, David A.; Cole, David K.; Godkin, Andrew

doi:10.4049/jimmunol.1402787

Cited by 13 publications

(14 citation statements)

References 65 publications

(82 reference statements)

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“…The use of MHC class II tetramers presents considerable technical challenges (52), but we showed that sensitive tetramer reagents could be generated by identifying immunodominant epitopes and their HLA restriction in this system. In BAL, most RSV-specific CD4 + T cells expressed the lectin CD69, which in humans is the only well-recognized marker of CD4 + Trm cells (53).…”

Section: Discussionmentioning

confidence: 98%

Epitope-specific airway-resident CD4+ T cell dynamics during experimental human RSV infection

Guvenel

Jóźwik

Ascough

et al. 2019

Journal of Clinical Investigation

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BACKGROUNDRespiratory syncytial virus (RSV) is an important cause of acute pulmonary disease and one of the last remaining major infections of childhood for which there is no vaccine. CD4+ T cells play a key role in antiviral immunity, but they have been little studied in the human lung.METHODSHealthy adult volunteers were inoculated i.n. with RSV A Memphis 37. CD4+ T cells in blood and the lower airway were analyzed by flow cytometry and immunohistochemistry. Bronchial soluble mediators were measured using quantitative PCR and MesoScale Discovery. Epitope mapping was performed by IFN-γ ELISpot screening, confirmed by in vitro MHC binding.RESULTSActivated CD4+ T cell frequencies in bronchoalveolar lavage correlated strongly with local C-X-C motif chemokine 10 levels. Thirty-nine epitopes were identified, predominantly toward the 3′ end of the viral genome. Five novel MHC II tetramers were made using an immunodominant EFYQSTCSAVSKGYL (F-EFY) epitope restricted to HLA-DR4, -DR9, and -DR11 (combined allelic frequency: 15% in Europeans) and G-DDF restricted to HLA-DPA1*01:03/DPB1*02:01 and -DPA1*01:03/DPB1*04:01 (allelic frequency: 55%). Tetramer labeling revealed enrichment of resident memory CD4+ T (Trm) cells in the lower airway; these Trm cells displayed progressive differentiation, downregulation of costimulatory molecules, and elevated CXCR3 expression as infection evolved.CONCLUSIONSHuman infection challenge provides a unique opportunity to study the breadth of specificity and dynamics of RSV-specific T-cell responses in the target organ, allowing the precise investigation of Trm recognizing novel viral antigens over time. The new tools that we describe enable precise tracking of RSV-specific CD4+ cells, potentially accelerating the development of effective vaccines.TRIAL REGISTRATIONClinicalTrials.gov NCT02755948.FUNDINGMedical Research Council, Wellcome Trust, National Institute for Health Research.

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Section: Discussionmentioning

confidence: 98%

Epitope-specific airway-resident CD4+ T cell dynamics during experimental human RSV infection

Guvenel

Jóźwik

Ascough

et al. 2019

Journal of Clinical Investigation

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“…T-cell clones, DCD10, and ILA1 were created ( 30 , 31 ) and passaged as previously described ( 32 ). Briefly, clones were expanded with irradiated (3,100 Gy) PBMCs from three donors in R10 [RPMI 1640 supplemented with 10% FBS, 100 U/ml penicillin, 100 µg/ml streptomycin, 1× MEM non-essential amino acid, 1 mM sodium pyruvate, 10 mM HEPES buffer (Life Technology)] with 20 IU/ml of IL-2 (Aldesluekin, Proleukin, Prometheus, San Diego, CA, USA) and 1 µg/ml of phytohaemaglutinin (Alere, Cheshire, UK).…”

Section: Methodsmentioning

confidence: 99%

Metabolic Adaptation of Human CD4+ and CD8+ T-Cells to T-Cell Receptor-Mediated Stimulation

et al. 2017

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Linking immunometabolic adaptation to T-cell function provides insight for the development of new therapeutic approaches in multiple disease settings. T-cell activation and downstream effector functions of CD4+ and CD8+ T-cells are controlled by the strength of interaction between the T-cell receptor (TCR) and peptides presented by human leukocyte antigens (pHLA). The role of TCR–pHLA interactions in modulating T-cell metabolism is unknown. Here, for the first time, we explore the relative contributions of the main metabolic pathways to functional responses in human CD4+ and CD8+ T-cells. Increased expression of hexokinase II accompanied by higher basal glycolysis is demonstrated in CD4+ T-cells; cytokine production in CD8+ T-cells is more reliant on oxidative phosphorylation. Using antigen-specific CD4+ and CD8+ T-cell clones and altered peptide ligands, we demonstrate that binding affinity tunes the underlying metabolic shift. Overall, this study provides important new insight into how metabolic pathways are controlled during antigen-specific activation of human T-cells.

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“…In contrast to CD8 + cells, CD4 + T cell exhaustion is still poorly understood, which is mainly due to the lower ex vivo frequency of peripheral blood virus-specific CD4 + T cells during chronic infection as well as technical limitations, particularly in generating MHC class II/antigen complexes by recombinant methods, and lower affinity binding of CD4 + TCR to cognate epitope on MHC class II molecule [58,59]. To date, it has been shown that exhausted CD4 + T cells lose the ability to secrete TNF-α, IFN-γ and IL-2, but increase production of IL-10 and IL-21 [60,61].…”

Section: T Cell Exhaustion In Hcv Infectionmentioning

confidence: 99%

Reversal of T Cell Exhaustion in Chronic HCV Infection

Osuch

Metzner

Cortés

2020

Viruses

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The long-term consequences of T cell responses’ impairment in chronic HCV infection are not entirely characterized, although they may be essential in the context of the clinical course of infection, re-infection, treatment-mediated viral clearance and vaccine design. Furthermore, it is unclear whether a complete reinvigoration of HCV-specific T cell response may be feasible. In most studies, attempting to reverse the effects of compromised immune response quality by specific blockades of negative immune regulators, a restoration of functional competence of HCV-specific T cells was shown. This implies that HCV-induced immune dysfunction may be reversible. The advent of highly successful, direct-acting antiviral treatment (DAA) for chronic HCV infection instigated investigation whether the treatment-driven elimination of viral antigens restores T cell function. Most of studies demonstrated that DAA treatment may result in at least partial restoration of T cell immune function. They also suggest that a complete restoration comparable to that seen after spontaneous viral clearance may not be attained, pointing out that long-term antigenic stimulation imprints an irreversible change on the T cell compartment. Understanding the mechanisms of HCV-induced immune dysfunction and barriers to immune restoration following viral clearance is of utmost importance to diminish the possible long-term consequences of chronic HCV infection.

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Enhanced Detection of Antigen-Specific CD4+ T Cells Using Altered Peptide Flanking Residue Peptide–MHC Class II Multimers

Cited by 13 publications

References 65 publications

Epitope-specific airway-resident CD4+ T cell dynamics during experimental human RSV infection

Epitope-specific airway-resident CD4+ T cell dynamics during experimental human RSV infection

Metabolic Adaptation of Human CD4+ and CD8+ T-Cells to T-Cell Receptor-Mediated Stimulation

Reversal of T Cell Exhaustion in Chronic HCV Infection

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