Fig. 7: Deficiency of Adam15 in CD8+ T cells and macrophages protects the cells from CS-induced activation of the mitochondrial apoptosis pathway in vivo and in vitro. | Mucosal Immunology

Fig. 7: Deficiency of Adam15 in CD8+ T cells and macrophages protects the cells from CS-induced activation of the mitochondrial apoptosis pathway in vivo and in vitro.

From: A disintegrin and metalloproteinase domain-15 deficiency leads to exaggerated cigarette smoke-induced chronic obstructive pulmonary disease (COPD)-like disease in mice

Fig. 7

a Alveolar macrophages (AMs) were isolated from the lungs of WT and Adam15−/− −/− mice that had been exposed to air or CS for 1 week or 3 months (Mo) using BAL. b CD8+ T cells were isolated from the lungs of WT and Adam15−/− mice exposed to air or CS for 1 month as described in “Methods”. a, b Immediately after the AMs and CD8+ T cells were isolated they were fixed and then immunostained for intracellular active caspase-3. Staining was quantified, as described in the “Methods”. Active caspase-3 was not detected in macrophages or CD8+ T cells isolated from air-exposed WT or Adam15−/− mice. Data are mean ± SEM; n = 5 mice/group in (a), and n = 4–5 mice/group in (b). Data were analyzed using a one-way ANOVA followed by pair-wise testing with two-tailed Student’s t tests. *P < 0.05 versus the group indicated. c AMs were isolated from the lungs of WT and Adam15−/− mice exposed to air or CS for 1 month using BAL. Activation of the mitochondrial apoptosis pathway was measured as loss of mitochondrial membrane potential (lack of staining of the cells with the mitochondrial dye, JC-1 which stains only viable mitochondria). Data are mean ± SEM; n = 5 mice/group. Data were analyzed using a one-way ANOVA followed by pair-wise testing with two-tailed Student’s t tests. *P < 0.05 versus the group indicated. d AMs isolated from naïve WT and Adam15−/− mice were incubated at 37 °C for up to 24 h with or without 10% CS extract (CSE). e CD8+ T cells isolated from the spleens of naïve WT Adam15−/− mice were incubated at 37 °C for up to 24 h with or without 5% CSE. d, e Activation of the mitochondrial apoptosis pathway was measured as loss of mitochondrial membrane potential (lack of staining of the cells with JC-1). Data are mean ± SEM; n = 3 separate experiments in (d, e). Data were analyzed using a one-way ANOVA followed by pair-wise testing with two-tailed Student’s t tests. *P < 0.05 versus the group indicated.

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