Abstract
Polymorphonuclear leukocytes (PMN) play a prominent role in host defense in mammals against invading bacteria (1,2). Among the essential attributes of these highly specialized cells are the elaboration of an array of cytotoxic peptides and polypeptides that can be targeted at bacterial prey. This includes the bactericidal/permeability-increasing protein (BPI), a cytotoxic protein that at nM concentrations acts selectively against many Gram-negative bacteria. The principal determinant of the target-cell selectivity and potency of BPI is its ability to bind avidly to lipopolysaccarides (LPS), abundant glycolipids found uniquely in the outer leaflet of the outer membrane of these bacteria (3,4). Binding of BPI to bacterial outer membrane LPS not only initiates antibacterial cytotoxicity but also blocks the potent pro-inflammatory activity of this bacterial product (5). Host responses to LPS fuel the delivery of host defenses (e.g., PMN) to sites of infection but can also lead to profound inflammatory injury if inadequately regulated (4,6). By contributing to the eradication of the bacteria that produce LPS and by blocking the activity of LPS already present, BPI can play a major role in elimination of invading Gram-negative bacteria and in downregulating further host responses to these bacteria.
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References
Elsbach, P., Weiss, J., and Levy, O. (1999) Oxygen-independent antimicrobial systems of phagocytes, in Inflammation: Basic Principles and Clinical Correlates, 3rd ed. (Gallin, J. I., Snyderman, R. A., and Nathan, C. A., eds.), Raven Press, New York, pp. 801ā817.
Levy, O. (1996) Antibiotic proteins of polymorphonuclear leukocytes. Eur. J. Haematol. 56, 263ā277.
Gazzano-Santoro, H., Parent, J. B., Grinna, L., Horwitz, A., Parsons, T., Theofan, G., Elsbach, P., Weiss, J., and Conlon, P. J. (1992) High-affinity binding of the bactericidal/permeability-increasing protein and a recombinant amino-terminal fragment to the lipid A region of lipopolysaccharide. Infect. Immun. 60, 754ā761.
Elsbach, P. and Weiss, J. (1993) The bactericidal/permeability-increasing protein (BPI), a potent element in host-defense against gram-negative bacteria and lipolysaccharide. Immunobiology 187, 417ā429.
Weiss, J., Elsbach, P., Shu, C., Castillo, J., Grinna, L., Horwitz, A., and Theofan, G. (1992) Human bactericidal/permeability-increasing protein and a recombinant NH2-terminal fragment cause killing of serum-resistant gram-negative bacteria in whole blood and inhibit tumor necrosis factor release induced by the bacteria. J. Clin. Invest. 90, 1122ā1130.
Rietschel, E. T. and Brade, H. (1992) Bacterial endotoxins. Sci. Amer. 267, 54ā61.
Abrahamson, S., Wu, H., Williams, R., Der, K., Ottah, N., Little, R., Gazzano-Santoro, H., Theofan, G., Bauer, R., Leigh, S., et. al. (1997) Biochemical characterization of recombinant fusions of lipopolysaccharide binding protein and bactericidal/permeability-increasing protein. Implications in biological activity. J. Biol. Chem. 72, 2149ā2155.
Weiss, J., Elsbach, P., Olsson, I., and Odeberg, H. (1978) Purification and characterization of a potent bactericidal and membrane-active protein from the granules of human polymorphonuclear leukocytes. J. Biol. Chem. 253, 2664ā2672.
Elsbach, P., Weiss, J., Franson, R., Beckerdite-Quagliata, S., Schneider, A., and Harris, L. (1979) Separation and purification of a potent bactericidal/permeability-increasing protein and a closely related phospholipase A2 from rabbit poly-morphonuclear leukocytes. Observations on their relationship. J. Biol. Chem. 254, 11,000ā11,009.
Mannion, B. A., Kalatzis, E. S., Weiss, J., and Elsbach, P. (1989) Preferential binding of the neutrophil cytoplasmic granule-derived bactericidal/permeability-increasing protein to target bacteria. Implications and use as a means of purification. J. Immunol. 142, 2807ā2812.
Weiss, J. and Olsson, I. (1987) Cellular and subcellular localization of the bactericidal/permeability-increasing protein of neutrophils. Blood 69, 652ā659.
Ooi, C. E., Weiss, J., Levy, O., and Elsbach, P. (1990) Isolation of two isoforms of a novel 15-kDa protein from rabbit polymorphonuclear leukocytes that modulate the antibacterial actions of other leukocyte proteins. J. Biol. Chem. 265, 15,956ā15,962.
Weinrauch, Y., Foreman, A., Shu, C., Zarember, K., Levy, O., Elsbach, P., and Weiss, J. (1995) Extracellular accumulation of potently microbicidal bactericidal/permeability-increasing protein (BPI) and p15s in an evolving sterile rabbit peritoneal inflammatory exudate. J. Clin. Invest. 95, 1916ā1924.
Zarember, K., Elsbach, P., Kim, K., and Weiss, J. (1997) p15s (15-kD antimicrobial proteins) are stored in the secondary granules of rabbit granulocytes: Implications for antibacterial synergy with the bactericidal/permeability-increasing protein in inflammatory fluids. Blood 89, 672ā679.
Weiss, J., Beckerdite-Quagliata, S., and Elsbach, P. (1980) Resistance of gram-negative bacteria to purified leukocyte proteins: relation to binding and bacterial lipopolysaccharide structure. J. Clin. Invest. 65, 619ā628.
Capodici, C., Chen, S., Sidorczyk, Z., Elsbach, P., and Weiss, J. (1994) Effect of lipopolysaccharide (LPS) chain length on interactions of bactericidal/permeability-increasing protein and its bioactive 23-kilodalton NH2-terminal fragment with isolated LPS and intact Proteus mirabilis and Escherichia coli. Infect. Immun. 62, 259ā265.
Weiss, J., Victor, M., and Elsbach, P. (1983) Role of charge and hydrophobic interactions in the action of the bactericidal/permeability-increasing protein of neutrophils on gram-negative bacteria. J. Clin. Invest. 71, 540ā549.
Weiss, J., Victor, M., Cross, A., and Elsbach, P. (1982) Sensitivity of K1-encapsulated Escherichia coli to killing by the bactericidal/permeability-increasing protein of rabbit and human neutrophils. Infect. Immun. 38, 1149ā1153.
Elsbach, P., Weiss, J., and Kao, L. (1985) The role of intramembrane Ca2+ in the hydrolysis of the phospholipids of Escherichia coli by Ca2+-dependent phospholi-pases. J. Biol. Chem. 260, 1618ā1622.
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Weiss, J. (2000). Isolation of the Bactericidal/Permeability-Increasing Protein from Polymorphonuclear Leukocytes by Reversible Binding to Target Bacteria. In: Evans, T.J. (eds) Septic Shock Methods and Protocols. Methods in Molecular Medicineā¢, vol 36. Humana Press. https://doi.org/10.1385/1-59259-216-3:37
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DOI: https://doi.org/10.1385/1-59259-216-3:37
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