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. 2022 Mar 22;7(6):e154187.
doi: 10.1172/jci.insight.154187.

CMV-associated T cell and NK cell terminal differentiation does not affect immunogenicity of ChAdOx1 vaccination

Hannah R Sharpe  1 , Nicholas M Provine  2 , Georgina S Bowyer  1 , Pedro Moreira Folegatti  1 , Sandra Belij-Rammerstorfer  1 , Amy Flaxman  1 , Rebecca Makinson  1 , Adrian Vs Hill  1 , Katie J Ewer  1 , Andrew J Pollard  3 , Paul Klenerman  2 , Sarah Gilbert  1 , Teresa Lambe  1   3   4
Affiliations

CMV-associated T cell and NK cell terminal differentiation does not affect immunogenicity of ChAdOx1 vaccination

Hannah R Sharpe et al. JCI Insight. .

Abstract

Cytomegalovirus (CMV) is a globally ubiquitous pathogen with a seroprevalence of approximately 50% in the United Kingdom. CMV infection induces expansion of immunosenescent T cell and NK cell populations, with these cells demonstrating lower responsiveness to activation and reduced functionality upon infection and vaccination. In this study, we found that CMV+ participants had normal T cell responses after a single-dose or homologous vaccination with the viral vector chimpanzee adenovirus developed by the University of Oxford (ChAdOx1). CMV seropositivity was associated with reduced induction of IFN-γ-secreting T cells in a ChAd-Modified Vaccinia Ankara (ChAd-MVA) viral vector vaccination trial. Analysis of participants receiving a single dose of ChAdOx1 demonstrated that T cells from CMV+ donors had a more terminally differentiated profile of CD57+PD1+CD4+ T cells and CD8+ T cells expressing less IL-2Rα (CD25) and fewer polyfunctional CD4+ T cells 14 days after vaccination. NK cells from CMV-seropositive individuals also had a reduced activation profile. Overall, our data suggest that although CMV infection enhances immunosenescence of T and NK populations, it does not affect antigen-specific T cell IFN-γ secretion or antibody IgG production after vaccination with the current ChAdOx1 nCoV-19 vaccination regimen, which has important implications given the widespread use of this vaccine, particularly in low- and middle-income countries with high CMV seroprevalence.

Keywords: COVID-19; NK cells; T cells; Vaccines.

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Conflict of interest statement

Conflict of interest: The University of Oxford has a partnership with AstraZeneca for further development of ChAdOx1 nCoV-19. SG is a cofounder and board member of Vaccitech (collaborators in early development of this vaccine candidate), holds stock in this company, and is an inventor on a patent on use of ChAdOx1-vectored vaccines and a patent application on this SARS-CoV-2 vaccine (PCT/GB2012/000467, UK 2003670.3). TL is an inventor on a pending patent application (UK 2003670.3) on this SARS-CoV-2 vaccine and was a consultant to Vaccitech during this study. SG and TL are inventors on intellectual property (patent WO2021181100) covering ChAdOx1 nCoV-19 preclinical data. PMF was a consultant to Vaccitech during this study. AJP is chair of the UK Department of Health and Social Care’s Joint Committee on Vaccination and Immunisation but does not participate in policy advice on coronavirus vaccines and was a member of the WHO Strategic Advisory Group of Experts during this study. AJP is a National Institute for Health Research Senior Investigator. AVSH is a cofounder of and consultant to Vaccitech, is an inventor on a patent on the design and use of ChAdOx1-vectored vaccines (PCT/GB2012/000467), reports personal fees from Vaccitech, and might benefit from royalty income to the University of Oxford from sales of this vaccine and by AstraZeneca and sublicensees. HRS, SBR, and AF are contributors on intellectual property (patent WO2021181100) on ChAdOx1 nCoV-19 preclinical data.

Figures

Figure 1
Figure 1. T cell phenotype of trial participants when stratified by CMV serostatus.
(A) CD57+KLRG1+ CD4+ and CD8+ T cells in CMV+ and CMV– donors from ChAdOx1 S-D and ChAdOx1-MVA trial cohorts. (B) CD45RA/CCR7 memory profile of CD8+ T cells. (C) CD4+CD57+PD1+ T cells in CMV+ and CMV– individuals. (D) CD25 expression on CD8+ T cells stratified by CMV serostatus and pooled day 0 CD8+CD25+ T cells. ChAdOx1 S-D: n = 20 CMV seronegative, n = 6 CMV seropositive. ChAdOx1-MVA: n = 15 CMV seronegative, n = 4 CMV seropositive. Open circles = CMV seronegative, closed circles = CMV seropositive. Statistics conducted using Mann-Whitney U test and mixed effects analysis with Holm-Šidák multiple comparisons. *P < 0.05, **P < 0.01, ***P < 0.005, ****P < 0.0001. Error bars shown as median ± IQR.
Figure 2
Figure 2. T cell functionality in prime and prime-boost vaccine regimens when stratified by CMV serostatus.
(A) T cell IFN-γ production from groups 1 and 2 of the ChAdOx1-MVA P-B vaccine regimen measured by ELISPOT and stratified for CMV serostatus. (B) tIgG from ChAdOx1 S-D and ChAdOx1 P-B trial participants. (C) Polyfunctionality of CD4+ T cells from ChAdOx1 S-D participants at day 14 postvaccination, measuring expression of CD25, CD107a, IFN-γ, IL-2, and TNF. (D) ChAdOx1 S-D cohort and ChAdOx1-MVA cohort: percentage of cytokine+ T cells within the CD8+CD57+KLRG1+ population following vaccination. (E) T cell IFN-γ production from groups 1 and 2 of the ChAdOx1-MVA P-B vaccine regimen measured by ELISPOT. (F) Fold change of ELISPOT response compared with day 0 from pooled groups 1 and 2 of the ChAdOx2-MVA P-B vaccine regimen and fold change of ELISPOT IFN-γ production when compared with day 0, both stratified for CMV serostatus. ChAdOx1 S-D: n = 31 CMV seronegative, n = 13 CMV seropositive. ChAdOx1 P-B: n = 28 CMV seronegative, n = 20 CMV seropositive. ChAdOx1-MVA: n = 15 CMV seronegative, n = 4 CMV seropositive. Open circles = CMV seronegative, closed circles = CMV seropositive. Statistics conducted using mixed effects analysis with Holm-Šidák multiple comparisons. *P < 0.05, **P < 0.01. Error bars presented as median ± IQR. SFU, spot-forming units (per million cells).
Figure 3
Figure 3. NK cell phenotype with CMV serostatus.
(A) CD57+NKG2C+ NK cell frequency with CMV serostatus. (B) CD69 expression on NK cells stratified by CMV serostatus. (C) Frequency of CD25+ NK cells and CD57+CD25+ NK cells. (D) t-SNE analysis conducted on 26 ChAdOx1 cohort samples across 4 time points (day 0, day 7, day 14, and day 28). t-SNE plot was created by downsampling and concatenation of 25,000 randomly selected NK cells from each time point and sample. ChAdOx1 S-D: n = 20 CMV seronegative, n = 6 CMV seropositive. ChAdOx1-MVA: n = 15 CMV seronegative, n = 4 CMV seropositive. ChAd3-MVA: n = 8 CMV seronegative, n = 8 CMV seropositive. Open circles = CMV seronegative, closed circles = CMV seropositive. Statistics conducted using Mann-Whitney U test and mixed effects analysis with Holm-Šidák multiple comparisons. *P < 0.05, **P < 0.01, ***P < 0.005. Error bars presented as median ± IQR.
Figure 4
Figure 4. Changes in NK cell cytotoxicity profile with CMV positivity.
PBMCs from trial participants stimulated with relevant antigen peptides and stained for cytokine production and cytotoxicity. (A) NK cell cytotoxicity and cytokine production (IFN-γ, TNF, and CD107a) from the ChAdOx1 S-D cohort participants. (B) Correlation of antigen-specific CD4+ IL-2 secretion and NK cell activation (CD25) and proliferation (Ki-67) at day 14 after vaccination stratified by CMV serostatus. (C) NK cell cytotoxicity and cytokine production (IFN-γ, TNF, and CD107a) from ChAdOx1-MVA group 1+2 participants. (D) ChAd3-MVA participant NKG2C+ NK cell cytotoxicity and cytokine production (IFN-γ, granzyme B, and CD107a). ChAdOx1 S-D: n = 20 CMV seronegative, n = 6 CMV seropositive. ChAdOx1-MVA: n = 15 CMV seronegative, n = 4 CMV seropositive. ChAd3-MVA: n = 8 CMV seronegative, n = 8 CMV seropositive. Open circles = CMV seronegative, closed circles = CMV seropositive. Statistics conducted using Mann-Whitney U test, linear regression, and mixed effects analysis with Holm-Šidák multiple comparisons. *P < 0.05, **P < 0.01. Error bars shown as median ± IQR.
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