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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1993 Dec 15;90(24):12040–12044. doi: 10.1073/pnas.90.24.12040

In vivo protection against endotoxin by plasma high density lipoprotein.

D M Levine 1, T S Parker 1, T M Donnelly 1, A Walsh 1, A L Rubin 1
PMCID: PMC48121  PMID: 8265667

Abstract

Overwhelming bacterial infection is accompanied by fever, hypotension, disseminated intravascular coagulation, and multiple organ failure leading to death in 30-80% of cases. These classical symptoms of septic shock are caused by potent cytokines that are produced in response to endotoxin released from Gram-negative bacteria. Treatments with antibodies and receptor antagonists to block endotoxin or cytokine mediators have given mixed results in clinical trials. High density lipoprotein (HDL) is a natural component of plasma that is known to neutralize endotoxin in vitro. We report here that raising the plasma HDL concentration protects mice against endotoxin in vivo. Transgenic mice with 2-fold-elevated plasma HDL levels had more endotoxin bound to HDL, lower plasma cytokine levels, and improved survival rates compared with low-HDL mice. Intravenous infusion of HDL also protected mice, but only when given as reconstituted HDL prepared from phospholipid and either HDL apoprotein or an 18-amino acid peptide synthesized to mimic the structure of apolipoprotein A-I of HDL. Intact plasma HDL was mildly toxic, and HDL apoprotein was ineffective. The effectiveness of the reconstituted peptide renders very unlikely any significant contribution to protection by trace proteins in apo-HDL. These data suggest a simple leaflet insertion model for binding and neutralization of lipopolysaccharide by phospholipid on the surface of HDL. Plasma HDL may normally act to protect against endotoxin; this protection may be augmented by administration of reconstituted HDL or reconstituted peptides.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anantharamaiah G. M. Synthetic peptide analogs of apolipoproteins. Methods Enzymol. 1986;128:627–647. doi: 10.1016/0076-6879(86)28096-9. [DOI] [PubMed] [Google Scholar]
  2. Baumberger C., Ulevitch R. J., Dayer J. M. Modulation of endotoxic activity of lipopolysaccharide by high-density lipoprotein. Pathobiology. 1991;59(6):378–383. doi: 10.1159/000163681. [DOI] [PubMed] [Google Scholar]
  3. Beutler B., Milsark I. W., Cerami A. C. Passive immunization against cachectin/tumor necrosis factor protects mice from lethal effect of endotoxin. Science. 1985 Aug 30;229(4716):869–871. doi: 10.1126/science.3895437. [DOI] [PubMed] [Google Scholar]
  4. Bone R. C. The pathogenesis of sepsis. Ann Intern Med. 1991 Sep 15;115(6):457–469. doi: 10.7326/0003-4819-115-6-457. [DOI] [PubMed] [Google Scholar]
  5. Bonomo E. A., Swaney J. B. A rapid method for the synthesis of protein-lipid complexes using adsorption chromatography. J Lipid Res. 1988 Mar;29(3):380–384. [PubMed] [Google Scholar]
  6. Brasseur R., De Meutter J., Vanloo B., Goormaghtigh E., Ruysschaert J. M., Rosseneu M. Mode of assembly of amphipathic helical segments in model high-density lipoproteins. Biochim Biophys Acta. 1990 Apr 17;1043(3):245–252. doi: 10.1016/0005-2760(90)90023-q. [DOI] [PubMed] [Google Scholar]
  7. Cavaillon J. M., Fitting C., Haeffner-Cavaillon N., Kirsch S. J., Warren H. S. Cytokine response by monocytes and macrophages to free and lipoprotein-bound lipopolysaccharide. Infect Immun. 1990 Jul;58(7):2375–2382. doi: 10.1128/iai.58.7.2375-2382.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chajek-Shaul T., Hayek T., Walsh A., Breslow J. L. Expression of the human apolipoprotein A-I gene in transgenic mice alters high density lipoprotein (HDL) particle size distribution and diminishes selective uptake of HDL cholesteryl esters. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6731–6735. doi: 10.1073/pnas.88.15.6731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Donnelly T. M., Kelsey S. F., Levine D. M., Parker T. S. Control of variance in experimental studies of hyperlipidemia using the WHHL rabbit. J Lipid Res. 1991 Jul;32(7):1089–1098. [PubMed] [Google Scholar]
  10. Espevik T., Nissen-Meyer J. A highly sensitive cell line, WEHI 164 clone 13, for measuring cytotoxic factor/tumor necrosis factor from human monocytes. J Immunol Methods. 1986 Dec 4;95(1):99–105. doi: 10.1016/0022-1759(86)90322-4. [DOI] [PubMed] [Google Scholar]
  11. Flegel W. A., Wölpl A., Männel D. N., Northoff H. Inhibition of endotoxin-induced activation of human monocytes by human lipoproteins. Infect Immun. 1989 Jul;57(7):2237–2245. doi: 10.1128/iai.57.7.2237-2245.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Francone O. L., Gurakar A., Fielding C. Distribution and functions of lecithin:cholesterol acyltransferase and cholesteryl ester transfer protein in plasma lipoproteins. Evidence for a functional unit containing these activities together with apolipoproteins A-I and D that catalyzes the esterification and transfer of cell-derived cholesterol. J Biol Chem. 1989 Apr 25;264(12):7066–7072. [PubMed] [Google Scholar]
  13. Glomset J. A. High-density lipoproteins in human health and disease. Adv Intern Med. 1980;25:91–116. [PubMed] [Google Scholar]
  14. Gordon D. J., Rifkind B. M. High-density lipoprotein--the clinical implications of recent studies. N Engl J Med. 1989 Nov 9;321(19):1311–1316. doi: 10.1056/NEJM198911093211907. [DOI] [PubMed] [Google Scholar]
  15. Harris H. W., Grunfeld C., Feingold K. R., Rapp J. H. Human very low density lipoproteins and chylomicrons can protect against endotoxin-induced death in mice. J Clin Invest. 1990 Sep;86(3):696–702. doi: 10.1172/JCI114765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Harris H. W., Grunfeld C., Feingold K. R., Read T. E., Kane J. P., Jones A. L., Eichbaum E. B., Bland G. F., Rapp J. H. Chylomicrons alter the fate of endotoxin, decreasing tumor necrosis factor release and preventing death. J Clin Invest. 1993 Mar;91(3):1028–1034. doi: 10.1172/JCI116259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Harris R. I., Stone P. C., Stuart J. An improved chromogenic substrate endotoxin assay for clinical use. J Clin Pathol. 1983 Oct;36(10):1145–1149. doi: 10.1136/jcp.36.10.1145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  19. Mathison J. C., Ulevitch R. J. The clearance, tissue distribution, and cellular localization of intravenously injected lipopolysaccharide in rabbits. J Immunol. 1979 Nov;123(5):2133–2143. [PubMed] [Google Scholar]
  20. Mathison J. C., Wolfson E., Ulevitch R. J. Participation of tumor necrosis factor in the mediation of gram negative bacterial lipopolysaccharide-induced injury in rabbits. J Clin Invest. 1988 Jun;81(6):1925–1937. doi: 10.1172/JCI113540. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Matz C. E., Jonas A. Micellar complexes of human apolipoprotein A-I with phosphatidylcholines and cholesterol prepared from cholate-lipid dispersions. J Biol Chem. 1982 Apr 25;257(8):4535–4540. [PubMed] [Google Scholar]
  22. Miller G. J., Miller N. E. Plasma-high-density-lipoprotein concentration and development of ischaemic heart-disease. Lancet. 1975 Jan 4;1(7897):16–19. doi: 10.1016/s0140-6736(75)92376-4. [DOI] [PubMed] [Google Scholar]
  23. Munford R. S., Andersen J. M., Dietschy J. M. Sites of tissue binding and uptake in vivo of bacterial lipopolysaccharide-high density lipoprotein complexes: studies in the rat and squirrel monkey. J Clin Invest. 1981 Dec;68(6):1503–1513. doi: 10.1172/JCI110404. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Munford R. S., Dietschy J. M. Effects of specific antibodies, hormones, and lipoproteins on bacterial lipopolysaccharides injected into the rat. J Infect Dis. 1985 Jul;152(1):177–184. doi: 10.1093/infdis/152.1.177. [DOI] [PubMed] [Google Scholar]
  25. Munford R. S., Hall C. L., Dietschy J. M. Binding of Salmonella typhimurium lipopolysaccharides to rat high-density lipoproteins. Infect Immun. 1981 Dec;34(3):835–843. doi: 10.1128/iai.34.3.835-843.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Munford R. S., Hall C. L., Dietschy J. M. Binding of Salmonella typhimurium lipopolysaccharides to rat high-density lipoproteins. Infect Immun. 1981 Dec;34(3):835–843. doi: 10.1128/iai.34.3.835-843.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Pugin J., Schürer-Maly C. C., Leturcq D., Moriarty A., Ulevitch R. J., Tobias P. S. Lipopolysaccharide activation of human endothelial and epithelial cells is mediated by lipopolysaccharide-binding protein and soluble CD14. Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):2744–2748. doi: 10.1073/pnas.90.7.2744. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Raetz C. R., Ulevitch R. J., Wright S. D., Sibley C. H., Ding A., Nathan C. F. Gram-negative endotoxin: an extraordinary lipid with profound effects on eukaryotic signal transduction. FASEB J. 1991 Sep;5(12):2652–2660. doi: 10.1096/fasebj.5.12.1916089. [DOI] [PubMed] [Google Scholar]
  29. Romeo D., Hinckley A., Rothfield L. Reconstitution of a functional membrane enzyme system in a monomolecular film. II. Formation of a functional ternary film of lipopolysaccharide, phospholipid and transferase enzyme. J Mol Biol. 1970 Nov 14;53(3):491–501. doi: 10.1016/0022-2836(70)90079-3. [DOI] [PubMed] [Google Scholar]
  30. Rubin E. M., Ishida B. Y., Clift S. M., Krauss R. M. Expression of human apolipoprotein A-I in transgenic mice results in reduced plasma levels of murine apolipoprotein A-I and the appearance of two new high density lipoprotein size subclasses. Proc Natl Acad Sci U S A. 1991 Jan 15;88(2):434–438. doi: 10.1073/pnas.88.2.434. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Schumaker V. N., Puppione D. L. Sequential flotation ultracentrifugation. Methods Enzymol. 1986;128:155–170. doi: 10.1016/0076-6879(86)28066-0. [DOI] [PubMed] [Google Scholar]
  32. Schumann R. R., Leong S. R., Flaggs G. W., Gray P. W., Wright S. D., Mathison J. C., Tobias P. S., Ulevitch R. J. Structure and function of lipopolysaccharide binding protein. Science. 1990 Sep 21;249(4975):1429–1431. doi: 10.1126/science.2402637. [DOI] [PubMed] [Google Scholar]
  33. Shemer R., Walsh A., Eisenberg S., Breslow J. L., Razin A. Tissue-specific methylation patterns and expression of the human apolipoprotein AI gene. J Biol Chem. 1990 Jan 15;265(2):1010–1015. [PubMed] [Google Scholar]
  34. Tobias P. S., McAdam K. P., Soldau K., Ulevitch R. J. Control of lipopolysaccharide-high-density lipoprotein interactions by an acute-phase reactant in human serum. Infect Immun. 1985 Oct;50(1):73–76. doi: 10.1128/iai.50.1.73-76.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Tobias P. S., Soldau K., Kline L., Lee J. D., Kato K., Martin T. P., Ulevitch R. J. Cross-linking of lipopolysaccharide (LPS) to CD14 on THP-1 cells mediated by LPS-binding protein. J Immunol. 1993 Apr 1;150(7):3011–3021. [PubMed] [Google Scholar]
  36. Ulevitch R. J., Johnston A. R. The modification of biophysical and endotoxic properties of bacterial lipopolysaccharides by serum. J Clin Invest. 1978 Dec;62(6):1313–1324. doi: 10.1172/JCI109252. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Ulevitch R. J., Johnston A. R., Weinstein D. B. New function for high density lipoproteins. Isolation and characterization of a bacterial lipopolysaccharide-high density lipoprotein complex formed in rabbit plasma. J Clin Invest. 1981 Mar;67(3):827–837. doi: 10.1172/JCI110100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Van Lenten B. J., Fogelman A. M., Haberland M. E., Edwards P. A. The role of lipoproteins and receptor-mediated endocytosis in the transport of bacterial lipopolysaccharide. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2704–2708. doi: 10.1073/pnas.83.8.2704. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Vanloo B., Morrison J., Fidge N., Lorent G., Lins L., Brasseur R., Ruysschaert J. M., Baert J., Rosseneu M. Characterization of the discoidal complexes formed between apoA-I-CNBr fragments and phosphatidylcholine. J Lipid Res. 1991 Aug;32(8):1253–1264. [PubMed] [Google Scholar]
  40. Victorov A. V., Medvedeva N. V., Gladkaya E. M., Morozkin A. D., Podrez E. A., Kosykh V. A., Yurkiv V. A. Composition and structure of lipopolysaccharide-human plasma low density lipoprotein complex. Analytical ultracentrifugation, 31P-NMR, ESR and fluorescence spectroscopy studies. Biochim Biophys Acta. 1989 Aug 21;984(1):119–127. doi: 10.1016/0005-2736(89)90351-9. [DOI] [PubMed] [Google Scholar]
  41. Walsh A., Ito Y., Breslow J. L. High levels of human apolipoprotein A-I in transgenic mice result in increased plasma levels of small high density lipoprotein (HDL) particles comparable to human HDL3. J Biol Chem. 1989 Apr 15;264(11):6488–6494. [PubMed] [Google Scholar]
  42. Weiser M. M., Rothfield L. The reassociation of lipopolysaccharide, phospholipid, and transferase enzymes of the bacterial cell envelope. Isolation of binary and ternary complexes. J Biol Chem. 1968 Mar 25;243(6):1320–1328. [PubMed] [Google Scholar]
  43. Wollenweber H. W., Morrison D. C. Synthesis and biochemical characterization of a photoactivatable, iodinatable, cleavable bacterial lipopolysaccharide derivative. J Biol Chem. 1985 Dec 5;260(28):15068–15074. [PubMed] [Google Scholar]
  44. Wright S. D., Ramos R. A., Tobias P. S., Ulevitch R. J., Mathison J. C. CD14, a receptor for complexes of lipopolysaccharide (LPS) and LPS binding protein. Science. 1990 Sep 21;249(4975):1431–1433. doi: 10.1126/science.1698311. [DOI] [PubMed] [Google Scholar]

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