CD95 engagement induces disseminated endothelial cell apoptosis in vivo: immunopathologic implications by Anne Janin, Christophe Deschaumes, Marjan Daneshpouy, Jérôme Estaquier, Juliette Micic-Polianski, Premavathy Rajagopalan-Levasseur, Khadija Akarid, Nicolas Mounier, Eliane Gluckman, Gérard Socié, and Jean Claude Ameisen Blood Volume 99(8):2940-2947 April 15, 2002 ©2002 by American Society of Hematology

Systemic CD95 engagement induces disseminated vascular endothelial cell death.(A) The TUNEL assay shows apoptosis (brown staining) of vascular endothelial cells (filled arrowheads) and additional cell types in tissue sections from liver, heart, kidneys, and... Systemic CD95 engagement induces disseminated vascular endothelial cell death.(A) The TUNEL assay shows apoptosis (brown staining) of vascular endothelial cells (filled arrowheads) and additional cell types in tissue sections from liver, heart, kidneys, and brain of wild-type (WT) mice killed 2 hours after injection of the JO2 CD95-specific antibody, whereas no apoptotic lesions were detected in CD95-defectivelpr (LPR) mice killed 6 hours after JO2 injection (the unstained endothelial cells are indicated by open arrowheads). (B) Endocardial lesions (heart) and subarachnoidal hemorrhage suffusions (brain), as well as typical electron microscopy features of endothelial cell apoptosis (kidney) were observed in wt mice killed 2 hours after JO2 injection (filled arrowheads). Anne Janin et al. Blood 2002;99:2940-2947 ©2002 by American Society of Hematology

Quantification of vascular lesions induced by systemic CD95 engagement Quantification of vascular lesions induced by systemic CD95 engagement.Wt mice were injected with JO2 (1) (n = 9), JO2 plus caspase-8 inhibitor (2) (n = 4), control immunoglobulin (3) (n = 4), LPS (> 150-fold excess of LPS content in JO2) (4) (n = 4), smCD9... Quantification of vascular lesions induced by systemic CD95 engagement.Wt mice were injected with JO2 (1) (n = 9), JO2 plus caspase-8 inhibitor (2) (n = 4), control immunoglobulin (3) (n = 4), LPS (> 150-fold excess of LPS content in JO2) (4) (n = 4), smCD95L (5) (n = 4), smCD95L plus caspase-8 inhibitor (6) (n = 4), sCD95L (7) (n = 5), the anti-FLAG antibody used to multimerize the sCD95L (8) (n = 5), or remained untreated (9) (n = 5); JO2 was injected in lpr mice (10) (n = 7), in gld mice (11) (n = 7), or in TNF-α−/−–Ltα−/−mice (12) (n = 4). Percentages are mean ± SD of 2 independent counts on tissue sections from liver, heart, kidneys, and brain of(n) different mice, as described in “Materials and methods.” *P < .007 compared to 1; °P < .03 compared to 5;+ P < .002 compared to 1, 11, and 12 (Wilcoxon test). Anne Janin et al. Blood 2002;99:2940-2947 ©2002 by American Society of Hematology

Systemic allogeneic immune responses induce disseminated vascular endothelial cell death.(A) The TUNEL assay shows apoptosis (brown staining) of vascular endothelial cells (filled arrowheads) and additional cell types in tissue sections from liver, heart, k... Systemic allogeneic immune responses induce disseminated vascular endothelial cell death.(A) The TUNEL assay shows apoptosis (brown staining) of vascular endothelial cells (filled arrowheads) and additional cell types in tissue sections from liver, heart, kidney, and brain of immunodeficient SCID recipient mice killed 2 days after transfer of 108lymphocytes from allogeneic wild type (WT) mice or from allogeneic CD95-defective lpr (LPR) mice, whereas no apoptotic lesions were detected in SCID recipient mice 2 days after transfer of 108 lymphocytes from allogeneic CD95L-defectivegld (GLD) mice (the unstained endothelial cells are indicated by open arrowheads). (B) Endocardial lesions (heart) and subarachnoidal hemorrhage suffusions (brain), as well as typical electron microscopy features of endothelial cell apoptosis (kidney) were observed in SCID recipients killed 2 days after transfer of 108 lymphocytes from allogeneic wt mice (filled arrowheads). Anne Janin et al. Blood 2002;99:2940-2947 ©2002 by American Society of Hematology

Quantification of vascular lesions induced by systemic allogeneic immune responses.SCID recipient mice were killed 2 days after transfer of 108 lymphocytes from syngeneic wt mice (1) (n = 4), or from allogeneic wt mice in the absence (2) (n = 9) or presence... Quantification of vascular lesions induced by systemic allogeneic immune responses.SCID recipient mice were killed 2 days after transfer of 108 lymphocytes from syngeneic wt mice (1) (n = 4), or from allogeneic wt mice in the absence (2) (n = 9) or presence of recipient treatment with caspase-8 inhibitor (3) (n = 4), or after transfer of 108 lymphocytes from allogeneicgld mice (4) (n = 4), or from allogeneic lprmice (5) (n = 4); SCID recipients were killed 14 days after transfer of 2.5 × 107 lymphocytes from syngeneic (6) (n = 4) or allogeneic wt mice (7) (n = 5). Percentages are mean ± SD of 2 independent counts on tissue sections from liver, heart, kidneys, and brain of (n) different mice, as described in “Materials and methods.” *P < .007 compared to 2; °P < .007 compared to 2 and 5 (Wilcoxon test). Anne Janin et al. Blood 2002;99:2940-2947 ©2002 by American Society of Hematology