title: Targeting immune co-stimulatory effects of PD-L1 and PD-L2 might represent an effective therapeutic strategy in stroke.
abstract: Stroke outcome is worsened by the infiltration of inflammatory immune cells into ischemic brains. Our recent study demonstrated that PD-L1- and to a lesser extent PD-L2-deficient mice had smaller brain infarcts and fewer brain-infiltrating cells vs. wild-type (WT) mice, suggesting a pathogenic role for PD-ligands in experimental stroke. We sought to ascertain PD-L1 and PD-L2-expressing cell types that affect T-cell activation, post-stroke in the context of other known co-stimulatory molecules. Thus, cells from male WT and PD-L-deficient mice undergoing 60 min of middle cerebral artery occlusion (MCAO) followed by 96 h of reperfusion were treated with neutralizing antibodies to study co-stimulatory and co-inhibitory interactions between CD80, cytotoxic T-lymphocyte antigen-4 (CTLA-4), PD-1, and PD-Ls that regulate CD8(+) and CD4(+) T-cell activation. We found that antibody neutralization of PD-1 and CTLA-4 signaling post-MCAO resulted in higher proliferation in WT CD8(+) and CD4(+) T-cells, confirming an inhibitory role of PD-1 and CTLA-4 on T-cell activation. Also, CD80/CD28 interactions played a prominent regulatory role for the CD8(+) T-cells and the PD-1/PD-L2 interactions were dominant in controlling the CD4(+) T-cell responses in WT mice after stroke. A suppressive phenotype in PD-L1-deficient mice was attributed to CD80/CTLA-4 and PD-1/PD-L2 interactions. PD-L2 was crucial in modulating CD4(+) T-cell responses, whereas PD-L1 regulated both CD8(+) and CD4(+) T-cells. To establish the contribution of PD-L1 and PD-L2 on regulatory B-cells (Bregs), infarct volumes were evaluated in male PD-L1- and PD-L2-deficient mice receiving IL-10(+) B-cells 4h post-MCAO. PD-L2- but not PD-L1-deficient recipients of IL-10(+) B-cells had markedly reduced infarct volumes, indicating a regulatory role of PD-L2 on Bregs. These results imply that PD-L1 and PD-L2 differentially control induction of T- and Breg-cell responses after MCAO, thus suggesting that selective targeting of PD-L1 and PD-L2 might represent a valuable therapeutic strategy in stroke.
authors.affiliation: Neuroimmunology Research, Portland Veterans Affairs Medical Center Portland, OR, USA ; Department of Neurology, Oregon Health and Science University Portland, OR, USA.
authors.forename: Sheetal
authors.initials: S
authors.lastname: Bodhankar
authors.affiliation: Department of Anesthesiology and Perioperative Medicine, Oregon Health and Science University Portland, OR, USA.
authors.forename: Yingxin
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Neuroimmunology Research, Portland Veterans Affairs Medical Center Portland, OR, USA ; Department of Neurology, Oregon Health and Science University Portland, OR, USA.
authors.forename: Andrew
authors.initials: A
authors.lastname: Lapato
authors.affiliation: Neuroimmunology Research, Portland Veterans Affairs Medical Center Portland, OR, USA ; Department of Neurology, Oregon Health and Science University Portland, OR, USA ; Department of Molecular Microbiology and Immunology, Oregon Health and Science University Portland, OR, USA.
authors.forename: Arthur A
authors.initials: AA
authors.lastname: Vandenbark
authors.affiliation: Department of Anesthesiology and Perioperative Medicine, Oregon Health and Science University Portland, OR, USA.
authors.forename: Stephanie J
authors.initials: SJ
authors.lastname: Murphy
authors.affiliation: Neuroimmunology Research, Portland Veterans Affairs Medical Center Portland, OR, USA ; Department of Neurology, Oregon Health and Science University Portland, OR, USA ; Department of Anesthesiology and Perioperative Medicine, Oregon Health and Science University Portland, OR, USA.
authors.forename: Halina
authors.initials: H
authors.lastname: Offner
keywords: MCAO; T-cells; co-stimulatory pathway; programmed death ligand-1 and 2; regulatory B cells