Epitope-specific airway-resident CD4+ T cell dynamics during experimental human RSV infection
Aleks Guvenel, … , Peter J. Openshaw, Christopher Chiu
Aleks Guvenel, … , Peter J. Openshaw, Christopher Chiu
Published December 9, 2019
Citation Information: J Clin Invest. 2020;130(1):523-538. https://doi.org/10.1172/JCI131696.
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Clinical Research and Public Health Immunology Infectious disease

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

Abstract

BACKGROUND Respiratory 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.METHODS Healthy 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.RESULTS Activated 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.CONCLUSIONS Human 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 REGISTRATION ClinicalTrials.gov NCT02755948.FUNDING Medical Research Council, Wellcome Trust, National Institute for Health Research.

Authors

Aleks Guvenel, Agnieszka Jozwik, Stephanie Ascough, Seng Kuong Ung, Suzanna Paterson, Mohini Kalyan, Zoe Gardener, Emma Bergstrom, Satwik Kar, Maximillian S. Habibi, Allan Paras, Jie Zhu, Mirae Park, Jaideep Dhariwal, Mark Almond, Ernie H.C. Wong, Annemarie Sykes, Jerico Del Rosario, Maria-Belen Trujillo-Torralbo, Patrick Mallia, John Sidney, Bjoern Peters, Onn Min Kon, Alessandro Sette, Sebastian L. Johnston, Peter J. Openshaw, Christopher Chiu

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Figure 8

Epitope-specific CD4+ T cells preferentially accumulate and express CXCR3 in the airway during acute RSV infection.

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Epitope-specific CD4+ T cells preferentially accumulate and express CXCR...
PBMCs from DPA1*01:03/DPB1*04:01–expressing individuals infected with RSV (n = 5) before infection and on day 10 were stained using DPB1*04:01/G-DDF tetramer and analyzed by flow cytometry. (A) FACS plots from 1 representative donor are shown, gated on CD3+ lymphocytes. (B) Cumulative data from blood (n = 11) and BAL (n = 7) are shown with P values for 2-tailed Mann-Whitney test comparing infected and uninfected groups, and 2-tailed Wilcoxon’s matched-pairs tests comparing time points. *P < 0.05, **P < 0.01, ***P < 0.001. PBMCs and BAL cells were costained with phenotypic markers of (C) T cell resident memory (CD69 and CD103), (D) proliferation/activation (Ki-67 and CD38), (E) memory subsets (CD45RA and CCR7), (F) costimulation (CD27 and CD28), homing receptors (G) CCR5 and CD62L and (H) CXCR3 and CCR4. FACS plots from 1 representative donor are shown, gated on CD3+CD4+ T lymphocytes. Red dots represent tetramer+ cells, and gray contours show total CD4+ T cells. The median frequencies of each subset are shown. Where not indicated with asterisks, differences were not statistically significant.