Liposome-mediated gene transfer to lung isografts

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Liposome-mediated gene transfer to lung isografts Carlos H.R. Boasquevisque, MDa, Teng C. Lee, BSa, Bassem N. Mora, MDa, David Peterson, BSb, William O. Osburn, BAc, Matthew Bernstein, BSb, Wei Zhang, BSb, Jennifer B. Nietupski, BSc, Ronald K. Scheule, PhDc, Joel D. Cooper, MDa, Mitchell D. Botney, MDb, G.Alexander Patterson, MDa The Journal of Thoracic and Cardiovascular Surgery Volume 114, Issue 5, Pages 783-792 (November 1997) DOI: 10.1016/S0022-5223(97)70082-5 Copyright © 1997 Mosby, Inc. Terms and Conditions

Fig. 1 Lung CAT activity at different times after intravenous administration. Each column corresponds to one animal. The bottom row represents nonacetylated chloramphenicol. The middle and top rows represent monoacetylated and diacetylated chloramphenicol, respectively. A, CAT activity is maximal as early as 6 hours after intravenous infusion. One animal in the 12-hour group lost the intravenous injection because of a technical failure. B, CAT activity is high for at least 8 days. The Journal of Thoracic and Cardiovascular Surgery 1997 114, 783-792DOI: (10.1016/S0022-5223(97)70082-5) Copyright © 1997 Mosby, Inc. Terms and Conditions

Fig. 1 Lung CAT activity at different times after intravenous administration. Each column corresponds to one animal. The bottom row represents nonacetylated chloramphenicol. The middle and top rows represent monoacetylated and diacetylated chloramphenicol, respectively. A, CAT activity is maximal as early as 6 hours after intravenous infusion. One animal in the 12-hour group lost the intravenous injection because of a technical failure. B, CAT activity is high for at least 8 days. The Journal of Thoracic and Cardiovascular Surgery 1997 114, 783-792DOI: (10.1016/S0022-5223(97)70082-5) Copyright © 1997 Mosby, Inc. Terms and Conditions

Fig. 2 In situ hybridization performed with an 35S-radiolabeled CAT antisense cRNA probe. A, CAT expression occurred widely throughout the pulmonary interstitium, 48 hours after intravenous administration. B and C, CAT expression was also detected in cells lining the lumen of pulmonary vessels, presumably endothelial cells. The Journal of Thoracic and Cardiovascular Surgery 1997 114, 783-792DOI: (10.1016/S0022-5223(97)70082-5) Copyright © 1997 Mosby, Inc. Terms and Conditions

Fig. 2 In situ hybridization performed with an 35S-radiolabeled CAT antisense cRNA probe. A, CAT expression occurred widely throughout the pulmonary interstitium, 48 hours after intravenous administration. B and C, CAT expression was also detected in cells lining the lumen of pulmonary vessels, presumably endothelial cells. The Journal of Thoracic and Cardiovascular Surgery 1997 114, 783-792DOI: (10.1016/S0022-5223(97)70082-5) Copyright © 1997 Mosby, Inc. Terms and Conditions

Fig. 2 In situ hybridization performed with an 35S-radiolabeled CAT antisense cRNA probe. A, CAT expression occurred widely throughout the pulmonary interstitium, 48 hours after intravenous administration. B and C, CAT expression was also detected in cells lining the lumen of pulmonary vessels, presumably endothelial cells. The Journal of Thoracic and Cardiovascular Surgery 1997 114, 783-792DOI: (10.1016/S0022-5223(97)70082-5) Copyright © 1997 Mosby, Inc. Terms and Conditions

Fig. 3 CAT expression in lungs, livers, hearts, and kidneys 48 hours after intravenous administration (n = 3). Minimal CAT activity is present in livers and kidneys when compared with the CAT activity in lungs and hearts. The Journal of Thoracic and Cardiovascular Surgery 1997 114, 783-792DOI: (10.1016/S0022-5223(97)70082-5) Copyright © 1997 Mosby, Inc. Terms and Conditions

Fig. 4 CAT activity measured in lung isografts. A, Lungs transfected in vivo for 48 hours, serving as controls for B. B, Lung isografts transfected in vivo for 4 hours followed by transplantation and assessed on postoperative day 2. C, Lung isografts transfected in vivo for 6 hours, transplanted, and then assessed on postoperative day 2. D, Lungs transfected in vivo for 48 hours and assessed for CAT activity. This group was used as a control for group C. CAT activity was not significantly different between transplanted and control lungs. The Journal of Thoracic and Cardiovascular Surgery 1997 114, 783-792DOI: (10.1016/S0022-5223(97)70082-5) Copyright © 1997 Mosby, Inc. Terms and Conditions

Fig. 5 CAT activity was present in all grafts transfected ex vivo at 10° C with either 660 μg or 330 μg of DNA. A, Lung grafts transfected with 660 μg of DNA and 0.5 μmol of lipid 67:DOPE diluted in 10 ml of normal saline solution (n = 3). B, Lung grafts transfected with 330 μg of DNA and 0.25 μmol of lipid 67 diluted in 10 ml of normal saline solution (n = 3). The Journal of Thoracic and Cardiovascular Surgery 1997 114, 783-792DOI: (10.1016/S0022-5223(97)70082-5) Copyright © 1997 Mosby, Inc. Terms and Conditions

Fig. 5 CAT activity was present in all grafts transfected ex vivo at 10° C with either 660 μg or 330 μg of DNA. A, Lung grafts transfected with 660 μg of DNA and 0.5 μmol of lipid 67:DOPE diluted in 10 ml of normal saline solution (n = 3). B, Lung grafts transfected with 330 μg of DNA and 0.25 μmol of lipid 67 diluted in 10 ml of normal saline solution (n = 3). The Journal of Thoracic and Cardiovascular Surgery 1997 114, 783-792DOI: (10.1016/S0022-5223(97)70082-5) Copyright © 1997 Mosby, Inc. Terms and Conditions