An anti-inflammatory property of aprotinin detected at the level of leukocyte extravasation George Asimakopoulos, FRCSa, Richard Thompson, MRCPb, Sussan.

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Presentation transcript:

An anti-inflammatory property of aprotinin detected at the level of leukocyte extravasation George Asimakopoulos, FRCSa, Richard Thompson, MRCPb, Sussan Nourshargh, PhDb, Elaine A. Lidington, PhDb, Justin C. Mason, MRCPb, Chandana P. Ratnatunga, FRCSa, Dorian O. Haskard, FRCPb, Kenneth M. Taylor, FRCSa, R.Clive Landis, PhDb The Journal of Thoracic and Cardiovascular Surgery Volume 120, Issue 2, Pages 361-369 (August 2000) DOI: 10.1067/mtc.2000.106323 Copyright © 2000 American Association for Thoracic Surgery Terms and Conditions

Fig. 1 Aprotinin inhibits leukocyte extravasation but not leukocyte rolling or firm adhesion within rat mesenteric venules. Leukocyte endothelial cell responses were quantified during a 60-minute time course after fMLP application (10–7 mol/L) in randomly selected 100-μm mesenteric vessel segments from each animal. Aprotinin did not affect leukocyte rolling (A) or firm adhesion (B) but significantly attenuated transmigration through the vessel wall at 40 and 60 minutes after topical application of fMLP (C). Results are expressed as means ± SEM (n = 5 animals). *P <.05. The Journal of Thoracic and Cardiovascular Surgery 2000 120, 361-369DOI: (10.1067/mtc.2000.106323) Copyright © 2000 American Association for Thoracic Surgery Terms and Conditions

Fig. 2 Effect of aprotinin on leukocyte endothelial cell responses viewed by intravital microscopy. Photomicrographs of representative segments of postcapillary venule in rat mesentery are shown, as viewed by intravital microscopy. Both saline control (A) and aprotinin-treated animals (B) exhibited a basal leukocyte rolling flux before application of fMLP. Twenty minutes after fMLP, firm adhesion was evident in both control (C) and aprotinin-treated animals (D). Forty minutes after fMLP, leukocytes that had transmigrated were visible around the postcapillary venule (arrows) in control (E) but not aprotinin-treated animals (F) (bar = 50 μm). The Journal of Thoracic and Cardiovascular Surgery 2000 120, 361-369DOI: (10.1067/mtc.2000.106323) Copyright © 2000 American Association for Thoracic Surgery Terms and Conditions

Fig. 3 Aprotinin inhibits neutrophil transmigration across HUVECs in response to fMLP, IL-8, and PAF. Freshly isolated neutrophils from human venous blood were allowed to transmigrate in the presence of the indicated doses of aprotinin for 60 minutes through cultured endothelial cells on filters (3-μm pore size) in response to the following chemoattractants in the lower chamber: fMLP (10–9 mol/L, A ), IL-8 (10–9 mol/L, B ), and PAF (10–6 mol/L, C ). Neutrophils and HUVECs were pretreated with aprotinin for 15 and 60 minutes, respectively, before the addition of chemoattractant in the lower chamber. Results are expressed as mean percentage of cells transmigrated ± SEM from 5 experiments by using separate HUVEC isolates and peripheral blood donors. *P <.05; **P <.001 from chemoattractant without aprotinin. The Journal of Thoracic and Cardiovascular Surgery 2000 120, 361-369DOI: (10.1067/mtc.2000.106323) Copyright © 2000 American Association for Thoracic Surgery Terms and Conditions

Fig. 4 Aprotinin can target neutrophils or endothelial cells in isolation. Chemotactic responses to fMLP (A) and IL-8 (B) were carried out as described in the legend to Fig 3, pretreating either neutrophils (15 minutes) or HUVECs (60 minutes) alone or in combination with aprotinin at a dose of 1600 kIU/mL. Results shown are from representative experiments expressed as mean percentage of cells transmigrated ± SD. All assays were performed in duplicate. The Journal of Thoracic and Cardiovascular Surgery 2000 120, 361-369DOI: (10.1067/mtc.2000.106323) Copyright © 2000 American Association for Thoracic Surgery Terms and Conditions

Fig. 5 Aprotinin inhibits myeloperoxidase secretion from human neutrophils. Peripheral venous blood was incubated at room temperature for 60 minutes with fMLP (10–7 mol/L) in the presence and absence of aprotinin at the indicated doses. Plasma was collected, and immunoreactive myeloperoxidase was measured by means of enzyme-linked immunosorbent assay. All samples were measured in duplicate. Results are expressed as mean myeloperoxidase (in nanograms per milliliter) ± SEM (n = 5 donors). *P <.001 from fMLP without aprotinin. The Journal of Thoracic and Cardiovascular Surgery 2000 120, 361-369DOI: (10.1067/mtc.2000.106323) Copyright © 2000 American Association for Thoracic Surgery Terms and Conditions