NCMs regulate T cell survival in TLOs via PD-L1.

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NCMs regulate T cell survival in TLOs via PD-L1. NCMs regulate T cell survival in TLOs via PD-L1. (A to C) Increase of NCM–T cell contacts in TLOs in the PAT of mice 3 days after MI. (A) Representative TPLSM 3D reconstruction of PD-L1+ NCMs (red spheres) and T cells (cyan spheres) after MI (right) or sham (left) surgery. Examples of NCM–T cell contacts are indicated by the arrows. Scale bars, 30 μm. (B) TPLSM-derived quantification of PD-L1+ NCMs normalized on volume of TLOs. Data are shown as means ± SEM. n = 8 (sham) and n = 10 (MI) TLOs from three to five mice, respectively. **P < 0.01. (C) Percentage of PD-L1+ NCMs in direct contact with T cells in single TLOs. Data are shown as mean ± SEM. n = 9 (sham) and n = 10 (MI) TLOs from three to five mice, respectively. ***P < 0.001 (D) Representative single-plane TPLSM micrograph of a PD-L1+ (white) CD115+ (red) NCM in contact with CD3+ (green) T cells and B220+ (blue) B cells. PD-L1 at the NCM–T cell interface is indicated by the arrow. Scale bar, 10 μm. (E to J) Effect of treatment with control IgG or the anti–PD-L1 antibody on NCMs and T cells in TLOs of mice 3 days after MI. (E) Representative TPLSM reconstructions of CD115+ Ly6C− NCMs (red spheres) and T cell (cyan spheres). NCMs in direct contact with T cells are depicted as yellow spheres. Insets are shown in detail at the bottom. Scale bars, 80 μm. (F) Percentage of CD115+ Ly6C− NCMs in direct contact with T cells. Data are shown as means ± SEM. n = 7 (sham) and n = 9 (MI) TLOs from three mice per group. *P < 0.05. (G) Absolute quantification of CD115+ Ly6C− NCMs per volume of TLOs. Data are shown as means ± SEM. n = 7 (sham) and n = 9 (MI) TLOs from three mice per group. (H) Absolute quantification of T cells in single TLOs in the PAT of mice 3 days (3d; left) and 5 days (right) after MI and treatment. Data are shown as means ± SEM. n > 11 TLOs from three to four mice per group. *P < 0.05, **P < 0.01. (I) Representative TPLSM reconstructions of Annexin V+ (pink spheres) and Annexin V− (cyan spheres) T cells after treatment. Insets are shown in detail and as single colors at the bottom. Scale bars, 80 μm. (J) Percentage of Annexin V+ T cells in single TLOs 3 days after MI. Data are shown as means ± SEM. n > 11 TLOs from three to four mice per group. ***P < 0.001. Mariaelvy Bianchini et al. Sci. Immunol. 2019;4:eaar3054 Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works