The purposes of this application are to detail stimulus specific mechanisms of arachidonate metabolism and free radical and lysosomal enzyme release in human and sheep neutrophils (PMNs) and to test the hypothesis that immune vascular injury results from release of oxygen radicals from activated neutrophils adherent to endothelial cells. In vitro studies with PMNs isolated from human and sheep blood will determine activation responses with 4 different stimuli using aggregation, chemotaxis, superoxide anion release, lysosomal enzyme release, and metabolism of endogenous and exogenous arachidonic acid as indices of activation. In addition, detailed studies of changes in neutrophil morphology with each stimulus will be made. In vivo studies using chronically prepared unanesthetized sheep will determine the physiologic responses to the stimuli used to activate PMNs. morphologic studies will also be carried out in the sheep experiments to obtain evidence of PMN endothelial cell interactions. Drug treatment studies will be used to clarify mechanisms of release reactions. For each drug, dose response curves will be constructed for the PMN functions listed above using vehicle treated human and sheep cells as controls. Additional binding studies usingmpure C3a des arg and C5a des arg will be made to test the effects of drugs on anaphylatoxin binding. Similarly, measurements of PMN cAMP concentration will be made in those instances in which treatment alters release reactions. Studies with unanesthetized sheep will compare responses in vehicle treated and drug treated sheep. The results of these experiments will provide much needed data on the interrelationships among release reactions from activated PMNs and on the mechanisms of immune vascular injury. By comparing activation responses and mechanisms of release reactions in human and sheep pMNs, the extent to which measurements made in sheep may be applicable to immune vascular injury in humans will be clarified.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL036237-04
Application #
3351043
Study Section
Pathology A Study Section (PTHA)
Project Start
1986-08-01
Project End
1991-07-31
Budget Start
1989-08-01
Budget End
1990-07-31
Support Year
4
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Thomas Jefferson University
Department
Type
Schools of Medicine
DUNS #
061197161
City
Philadelphia
State
PA
Country
United States
Zip Code
19107
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Spath Jr, J A; Sloane, P J; Gee, M H et al. (1994) Loss of endothelium-dependent vasodilation in the pulmonary vessels of sheep after prolonged endotoxin. J Appl Physiol 76:361-9
Sloane, P J; Elsasser, T H; Spath Jr, J A et al. (1992) Plasma tumor necrosis factor-alpha during long-term endotoxemia in awake sheep. J Appl Physiol 73:1831-7
Doerr, T A; Rosolia, D L; Peters, S P et al. (1992) PGE1 inhibited PMN attachment to air emboli in vivo during infusion of ZAP without preventing lung injury. J Appl Physiol 72:340-51
Rosolia, D L; McKenna, P J; Gee, M H et al. (1992) Infusion of zymosan-activated plasma affects neutrophils in peripheral blood and bone marrow in sheep. J Leukoc Biol 52:501-15
Ishihara, Y; Rosolia, D L; McKenna, P J et al. (1990) Calcium is required for PMA induced superoxide release from human neutrophils. J Leukoc Biol 48:89-96
Peters, S P; Cerasoli Jr, F; Albertine, K H et al. (1990) ""Autoregulation"" of human neutrophil activation in vitro: regulation of phorbol myristate acetate-induced neutrophil activation by cell density. J Leukoc Biol 47:457-74
Cerasoli Jr, F; McKenna, P J; Rosolia, D L et al. (1990) Superoxide anion release from blood and bone marrow neutrophils is altered by endotoxemia. Circ Res 67:154-65
Albertine, K H; Cerasoli Jr, F; Tahamont, M V et al. (1989) Zymosan-activated plasma causes prolonged decreases in PMN superoxide release in sheep. J Appl Physiol 67:2481-90

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