Transition metal catalyzed formation of hydroxyl radical (OH) has been linked to neutrophil (PMN) microbicidal activity and tissue damage. Using spin trapping and other methodology, we and other found that PMN will not form OH unless provided with exogenous iron and that they possess two mechanisms (lactoferrin and myeloperoxidase [MPO] release) which limit their potential for OH production. These data suggest that the composite impact of exogenous catalysts, dynamics of the PMN oxidative and secretory response, and presence of factors capable of modulating that response should determine to a great extent the potential for in vivo generation of biologically significant quantities of OH. The investigation of all such possible factors is beyond the scope of any research program. Consequently, five hypotheses (specific aims) whose examination will provide major insight into the free radical biology of inflammation have been identified and will comprise Project 2. Surface adherent PMN and PMN """"""""primed"""""""" with different immunomodulatory factors exhibit alterations of cell functions.
Aim 1 examines oxygen-centered free radical formation of surface adherent PMNs as well as PMNs primed with lipopolysaccharide, y-interferon, interleukin-1, tumor necrosis factor (a and B), granulocyte-macrophage colony stimulating factor, and platelet activating factor. The endogenous capacity of these cells to generate OH as well as the dynamics of OH formation when they are provided with a catalyst will be examined using spin trapping and other recently described assays which appear to be specific for OH. MOst studies have examined PMN OH generation only in the context of an iron catalyst. Since copper can also act as an OH catalyst and is present in vivo, Aim 2 will examine the dynamics of OH generation by PMN stimulated in the presence of exogenous copper.
Aim 3 will explore the catalytic potential of several unique bacteria-derived iron chelates likely present in vivo at sites of infection. Four bacterial siderophores from Pseudomonas aeruginosa or Escherichia coli will be assessed for their ability to catalyze OH formation by both an enzymatic O2 generating system and PMN. In addition, the catalytic potential of Pseudomonas elastase induced cleavage products of transferrin will be assessed. MPO release limits of OH generation by iron supplemented PMN. Since MPO-deficient PMN are capable of killing some microbial pathogens, Aim 4 will test whether increased formation of OH (with microbial iron as the catalyst) is partially responsible for their retained microbicidal capacity. Eosinophils generate O2- and H2O2, lack lactoferrin, and possess a peroxidase (eosinophil peroxidase) distinct from MPO.
Aim 5 will determine the nature and dynamics of free radical generation by eosinophils. These results, will provide important data about the function of a related phagocyte and indirectly contribute to understanding of PMN function. Completion of these 5 aims will provide new insight into conditions necessary for in vivo generation of OH.

Project Start
Project End
Budget Start
Budget End
Support Year
2
Fiscal Year
1990
Total Cost
Indirect Cost
Name
University of Iowa
Department
Type
DUNS #
041294109
City
Iowa City
State
IA
Country
United States
Zip Code
52242
Britigan, B E; Rasmussen, G T; Olakanmi, O et al. (2000) Iron acquisition from Pseudomonas aeruginosa siderophores by human phagocytes: an additional mechanism of host defense through iron sequestration? Infect Immun 68:1271-5
McCormick, M L; Buettner, G R; Britigan, B E (1998) Endogenous superoxide dismutase levels regulate iron-dependent hydroxyl radical formation in Escherichia coli exposed to hydrogen peroxide. J Bacteriol 180:622-5
Miller, R A; Britigan, B E (1997) Role of oxidants in microbial pathophysiology. Clin Microbiol Rev 10:1-18
Olakanmi, O; Stokes, J B; Pathan, S et al. (1997) Polyvalent cationic metals induce the rate of transferrin-independent iron acquisition by HL-60 cells. J Biol Chem 272:2599-606
McCormick, M L; Metwali, A; Railsback, M A et al. (1996) Eosinophils from schistosome-induced hepatic granulomas produce superoxide and hydroxyl radical. J Immunol 157:5009-15
Nauseef, W M; Cogley, M; McCormick, S (1996) Effect of the R569W missense mutation on the biosynthesis of myeloperoxidase. J Biol Chem 271:9546-9
Miller, R A; Rasmussen, G T; Cox, C D et al. (1996) Protease cleavage of iron-transferrin augments pyocyanin-mediated endothelial cell injury via promotion of hydroxyl radical formation. Infect Immun 64:182-8
Andersen, K A; Britigan, B E; Wilson, M E (1996) Short report: regulation of inducible heat shock protein 70 genes in Leishmania chagasi. Am J Trop Med Hyg 54:471-4
Britigan, B E; Ratcliffe, H R; Buettner, G R et al. (1996) Binding of myeloperoxidase to bacteria: effect on hydroxyl radical formation and susceptibility to oxidant-mediated killing. Biochim Biophys Acta 1290:231-40
Edeker, B L; Rasmussen, G T; Britigan, B E (1995) Bicarbonate and phosphate ions protect transferrin from myeloperoxidase-mediated damage. J Leukoc Biol 58:59-64

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