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Volume 24, Issue 10, Pages 1873-1880 (October 2016)
Lentiviral Vector Gene Therapy Protects XCGD Mice From Acute Staphylococcus aureus Pneumonia and Inflammatory Response Giada Farinelli, Raisa Jofra Hernandez, Alice Rossi, Serena Ranucci, Francesca Sanvito, Maddalena Migliavacca, Chiara Brombin, Aleksandar Pramov, Clelia Di Serio, Chiara Bovolenta, Bernhard Gentner, Alessandra Bragonzi, Alessandro Aiuti Molecular Therapy Volume 24, Issue 10, Pages (October 2016) DOI: /mt Copyright © 2016 American Society of Gene & Cell Therapy Terms and Conditions
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Figure 1 Experimental plan. (a) XCGD and WT mice were injected intratracheally at 21 weeks, with S. aureus Newman strain at different doses (1 × 107 CFU and 2 × 107 colony-forming units (CFU)). Survival and weight changes were monitored daily. Surviving mice were euthanized 6 days after the infection and the lungs were harvested, homogenized, and cultured to determine the bacterial load. (b) Eight-week-old XCGD mice were treated with HSPCs transduced with different lentiviral vectors and bled at 14 and 18 weeks for DHR and VCN. At week 21, gene therapy-treated XCGD mice, XCGD and WT were injected intratracheally with S. aureus Newman strain (1 × 107 CFU). Survival and weight changes were monitored daily. Surviving mice were euthanized 5 days after the infection and broncho-alveolar lavage fluid was recovered. The lungs were harvested and either homogenized and cultured to determine the bacterial load and cytokine levels or subjected to histopathological analysis. Molecular Therapy , DOI: ( /mt ) Copyright © 2016 American Society of Gene & Cell Therapy Terms and Conditions
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Figure 2 Survival and weight loss in response to S. aureus Newman acute infection. XCGD, WT, and gene therapy-treated mice were injected intratracheally with S. aureus Newman strain (1 × 107 colony-forming units). (a) Survival was monitored over 5 days postinfection. Survival curves were displayed as Kaplan-Meyer curves. (b) Weight change was monitored over 5 days postinfection. Percentage loss of initial body weight is displayed. WT n = 15, XCGD n = 13, MSP.gp91_126T(2) n = 9, MSP.gp91 n = 5, BMT WT n = 7, XCGD MOCK n = 4. Molecular Therapy , DOI: ( /mt ) Copyright © 2016 American Society of Gene & Cell Therapy Terms and Conditions
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Figure 3 Bacterial load and broncho-alveolar lavage (BAL) cytology after S. aureus Newman acute infection. (a) The graph shows the bacterial load determined in the BAL fluid and in lung homogenate lungs. (b) The graph shows the content of total neutrophils, macrophages and lymphocytes recovered in the BAL fluid. Statistics shown in the figures represent the Kruskal Wallis test (nonparametric test with Dunns multiple comparison test; ***P < 0.001, **P < 0.05, *P < 0.01). Molecular Therapy , DOI: ( /mt ) Copyright © 2016 American Society of Gene & Cell Therapy Terms and Conditions
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Figure 4 Lung histopathological analysis following S. aureus infection. Images show hematoxylin- and eosin-stained lung sections of infected mice (5×). Scale bar = 200 microns. Molecular Therapy , DOI: ( /mt ) Copyright © 2016 American Society of Gene & Cell Therapy Terms and Conditions
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Figure 5 Cytokine and chemokine responses to S. aureus Newman acute infection. Cytokine and chemokine expression levels were assessed in lung homogenates obtained 5 days from infection. Results are shown as mean ± standard error of the mean. Statistics shown in the figures represent the Kruskal Wallis test (nonparametric test with Dunns multiple comparison test; *P < 0.01). Molecular Therapy , DOI: ( /mt ) Copyright © 2016 American Society of Gene & Cell Therapy Terms and Conditions
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