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. Author manuscript; available in PMC: 2021 May 1.
Published in final edited form as: Cytotherapy. 2020 Mar 29;22(5):276–290. doi: 10.1016/j.jcyt.2020.01.011

Figure 7: Expanded iNKT cells suppress activated CD4+ and CD8+ responder T cells in a contact-dependent manner.

Figure 7:

(A) Left panel, representative CD25 and CD45RA expression on gated CD4+ CFSE+ responder cells stimulated for 5 days. Black, unloaded bead stim; red, anti-CD2/CD3/CD28-loaded bead stim. Right panel, mean ± SEM percentage gated CD25+ activated cells among CD4+ responder cells (N = 4). Responder : iNKT cell ratios: 1:0, responders alone; 1:1, responders with iNKT cells in direct contact at 1:1; 1:1 TW, responders and iNKT cells at 1:1 but separated by transwell.

(B) Average divisions (left panel) and % Divided (right panel) for gated CFSE+CD4+CD25+ responder cells (N = 4) cultured with expanded iNKT cells, normalized to CD2/CD3/CD28-stimulated responders alone. Dashed line represents responders alone.

(C) Representative proliferation of CFSE+CD4+CD25+ responder cells. Green cross-hatch, generation 0; blue, generation 1; orange, generation 2. TW, representative proliferation plot with iNKT cells separated from responders via transwell.

(D) Left panel, representative CD25 and CD45RA expression on control CD8+ gated CFSE+ responder cells stimulated for 5 days. Black, unloaded bead stim; red, anti-CD2/CD3/CD28-loaded bead stim. Right panel, mean ± SEM percentage gated CD25+ activated cells among CD8+ responder cells (N = 4). Responder : iNKT cell ratios: 1:0, responders alone; 1:1, responders with iNKT cells in direct contact at 1:1; 1:1 TW, responders and iNKT cells at 1:1 but separated by transwell.

(E) Average divisions (left panel) and % Divided (right panel) for gated CFSE+CD8+CD25+ responder cells (N = 4) cultured with expanded iNKT cells, normalized to CD2/CD3/CD28-stimulated responders alone. Dashed line represents responders alone.

(F) Representative proliferation of CFSE+CD8+CD25+ responder cells. TW, representative proliferation plot with iNKT cells separated from responders via transwell. P < 0.05 is represented by *.