Fig. 4: Cellular response after BTI in PLWH.

A The functionality of SARS-CoV-2 Spike-specific T cells post-vaccination. Representative Dot plots of antigen-specific CD4 and CD8 T cells from vaccinated PLWH (n = 41, 4–5 months post-Dose 2). Antigen-specific T cells were identified by the expression of CD137⁺CD154⁺ for CD4 T cells and IFNγ/TNF for CD8 T cells following overnight stimulation with overlapping peptides coding for wild Type Spike (Wuhan) and HIV GAG (p24). Non-stimulated (NS) and Cytostim (CS) were used as negative and positive controls, respectively. B Functionality of SARS-CoV-2 Spike-specific T cells post-vaccination according to the immune status of PLWH, top. Frequency of Spike-specific T cells after the second dose of COVID-19 mRNA vaccine. PLWH were classified according to CD4 count at inclusion in the study. Antigen-specific T cells were gated as in A). The unspecific T-cell staining was removed as background from total Spike WT and HIV GAG response to display only virus-specific cellular response. Functionality of SARS-CoV-2-specific T cells after BTI, bottom. PLWH (red violin plots), COVID-19 convalescents (white violin plots), and vaccinated HDs (blue violin plots) were classified according to their history of SARS-CoV-2 infection and vaccination. Mann–Whitney two-tailed p-value with * for p < 0.05, ** for p < 0.01, *** for p < 0.001 and **** for p < 0.0001. C The breadth of SARS-CoV-2-specific T cells from PLWH BTI. Biplots represent the defined functionality of CD4 (top) and CD8 T cells (bottom) in response to Spike or MNO or GAG-derived peptides for each patient group (BTI PLWH, n = 16, 4–6 weeks post-BTI or COVID-19 convalescent, n = 18, 1-month post-infection). D Polyfunctionality of SARS-CoV-2-specific CD4 T cells in PLWH. SPICE analyses for in-depth functional profiling of SARS-CoV-2-specific CD4 T cells. Boolean gates were used to define the 8 possible combinations of IFNγ, TNF, and IL-2 on SARS-CoV-2-specific CD4 T cells identified by the upregulation of CD137 and CD154 after overnight stimulation with Spike- or non-Spike-derived peptides. SPICE pie charts represent the median frequency for each of the 8 possible functional profiles of SARS-CoV-2-specific CD4 T cells. The arcs indicate the proportion of cells that express IFNγ (red), TNF (green), and/or IL-2 (yellow-green). The antigen specificity and vaccination/infection status are indicated inside and above, respectively, for each pie chart. E Polyfunctionality of SARS-CoV-2-specific CD8 T cells in PLWH. SPICE analyses for in-depth functional profiling of SARS-CoV-2-specific CD8 T cells. Boolean gates were used to define the 32 possible combinations of IFNγ, TNF, Granzyme B, Perforin, and CD107a on SARS-CoV-2-specific CD8 T cells identified by the upregulation of IFNγ and/or TNF after overnight stimulation with Spike- or non-Spike-derived peptides. SPICE pie charts represent the median frequency for each of the 32 possible functional profiles of SARS-CoV-2-specific CD8 T cells. The arcs indicate the proportion of cells that express CD107a (green), Granzyme B (yellow), IFNγ (green), Perforin (blue), and/or TNF (purple). The antigen specificity and vaccination/infection status are indicated inside and above, respectively, for each pie chart. See also Supplementary Fig. 5.