Skip to main content
NCBI home page
Mediators of Inflammation logoLink to Mediators of Inflammation
. 1996 Feb;5(1):47–50. doi: 10.1155/S0962935196000087

Time-resolved fluoroimmunoassay for bactericidal/permeability-increasing protein

J-O Häggblom 1, A B Jokilammi-Siltanen 1, H Peuravuori 1, T J Nevalainen 1,✉
PMCID: PMC2365775  PMID: 18475697

Abstract

Bactericidal/permeability-increasing protein (BPI) is a cationic antimicrobial protein produced by polymorphonuclear leukocytes, that specifically interacts with and kills Gram-negative bacteria. BPl competes with lipopolysaccharide-binding protein (LBP) secreted by liver cells into blood plasma for binding to lipopolysaccharide (LPS) and thus reduces the proinflammatory effects of LPS. We have developed a time-resolved fluoroimmunoassay for BPI and measured the concentration of BPI in human serum and plasma samples. The assay is based on a rabbit antibody against recombinant BPI. This antibody specifically adheres to polymorphonuclear leukocytes in immunostained human tissues. The difference in the serum concentration of BPI between unselected hospitalized patients with and without an infection was statistically significant. The mean concentration of BPI in serum samples was 28.3 μg/l (range 1.64–132, S.D. 26.8, n = 83). In contrast, there was no difference between the two groups in the BPI levels in plasma samples. For all individuals tested, BPI levels were consistently higher in plasma samples compared to the matched serum samples. The mean concentration of BPI in plasma samples was 52.3 μg/l (range 0.9–403, S.D. 60.6, n = 90). There was a positive correlation between the concentration of BPI and the white blood cell count as well as between the BPI concentration and C-reactive protein (CRP) in serum samples. In conclusion, the present study demonstrates that BPI can be quantified reliably by time-resolved fluoroimmunoassay in human serum samples.

Full Text

The Full Text of this article is available as a PDF (751.4 KB).

Selected References

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

  1. Ammons W. S., Kung A. H. Recombinant amino terminal fragment of bactericidal/permeability-increasing protein prevents hemodynamic responses to endotoxin. Circ Shock. 1993 Nov;41(3):176–184. [PubMed] [Google Scholar]
  2. Corradin S. B., Heumann D., Gallay P., Smith J., Mauël J., Glauser M. P. Bactericidal/permeability-increasing protein inhibits induction of macrophage nitric oxide production by lipopolysaccharide. J Infect Dis. 1994 Jan;169(1):105–111. doi: 10.1093/infdis/169.1.105. [DOI] [PubMed] [Google Scholar]
  3. Elsbach P., Weiss J. Phagocytosis of bacteria and phospholipid degradation. Biochim Biophys Acta. 1988 Feb 24;947(1):29–52. doi: 10.1016/0304-4157(88)90018-4. [DOI] [PubMed] [Google Scholar]
  4. Fisher C. J., Jr, Marra M. N., Palardy J. E., Marchbanks C. R., Scott R. W., Opal S. M. Human neutrophil bactericidal/permeability-increasing protein reduces mortality rate from endotoxin challenge: a placebo-controlled study. Crit Care Med. 1994 Apr;22(4):553–558. doi: 10.1097/00003246-199404000-00008. [DOI] [PubMed] [Google Scholar]
  5. Forst S., Weiss J., Maraganore J. M., Heinrikson R. L., Elsbach P. Relation between binding and the action of phospholipases A2 on Escherichia coli exposed to the bactericidal/permeability-increasing protein of neutrophils. Biochim Biophys Acta. 1987 Aug 15;920(3):221–225. doi: 10.1016/0005-2760(87)90098-1. [DOI] [PubMed] [Google Scholar]
  6. Gazzano-Santoro H., Mészáros K., Birr C., Carroll S. F., Theofan G., Horwitz A. H., Lim E., Aberle S., Kasler H., Parent J. B. Competition between rBPI23, a recombinant fragment of bactericidal/permeability-increasing protein, and lipopolysaccharide (LPS)-binding protein for binding to LPS and gram-negative bacteria. Infect Immun. 1994 Apr;62(4):1185–1191. doi: 10.1128/iai.62.4.1185-1191.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gazzano-Santoro H., Parent J. B., Grinna L., Horwitz A., Parsons T., Theofan G., Elsbach P., Weiss J., Conlon P. J. High-affinity binding of the bactericidal/permeability-increasing protein and a recombinant amino-terminal fragment to the lipid A region of lipopolysaccharide. Infect Immun. 1992 Nov;60(11):4754–4761. doi: 10.1128/iai.60.11.4754-4761.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gray P. W., Flaggs G., Leong S. R., Gumina R. J., Weiss J., Ooi C. E., Elsbach P. Cloning of the cDNA of a human neutrophil bactericidal protein. Structural and functional correlations. J Biol Chem. 1989 Jun 5;264(16):9505–9509. [PubMed] [Google Scholar]
  9. Heumann D., Gallay P., Betz-Corradin S., Barras C., Baumgartner J. D., Glauser M. P. Competition between bactericidal/permeability-increasing protein and lipopolysaccharide-binding protein for lipopolysaccharide binding to monocytes. J Infect Dis. 1993 Jun;167(6):1351–1357. doi: 10.1093/infdis/167.6.1351. [DOI] [PubMed] [Google Scholar]
  10. Hsu S. M., Raine L., Fanger H. Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem. 1981 Apr;29(4):577–580. doi: 10.1177/29.4.6166661. [DOI] [PubMed] [Google Scholar]
  11. Ishii Y., Wang Y., Haziot A., del Vecchio P. J., Goyert S. M., Malik A. B. Lipopolysaccharide binding protein and CD14 interaction induces tumor necrosis factor-alpha generation and neutrophil sequestration in lungs after intratracheal endotoxin. Circ Res. 1993 Jul;73(1):15–23. doi: 10.1161/01.res.73.1.15. [DOI] [PubMed] [Google Scholar]
  12. Kohn F. R., Ammons W. S., Horwitz A., Grinna L., Theofan G., Weickmann J., Kung A. H. Protective effect of a recombinant amino-terminal fragment of bactericidal/permeability-increasing protein in experimental endotoxemia. J Infect Dis. 1993 Nov;168(5):1307–1310. doi: 10.1093/infdis/168.5.1307. [DOI] [PubMed] [Google Scholar]
  13. Marra M. N., Thornton M. B., Snable J. L., Wilde C. G., Scott R. W. Endotoxin-binding and -neutralizing properties of recombinant bactericidal/permeability-increasing protein and monoclonal antibodies HA-1A and E5. Crit Care Med. 1994 Apr;22(4):559–565. doi: 10.1097/00003246-199404000-00009. [DOI] [PubMed] [Google Scholar]
  14. Mathison J., Tobias P., Wolfson E., Ulevitch R. Regulatory mechanisms of host responsiveness to endotoxin (lipopolysaccharide). Pathobiology. 1991;59(3):185–188. doi: 10.1159/000163641. [DOI] [PubMed] [Google Scholar]
  15. Mészáros K., Parent J. B., Gazzano-Santoro H., Little R., Horwitz A., Parsons T., Theofan G., Grinna L., Weickmann J., Elsbach P. A recombinant amino terminal fragment of bactericidal/permeability-increasing protein inhibits the induction of leukocyte responses by LPS. J Leukoc Biol. 1993 Dec;54(6):558–563. doi: 10.1002/jlb.54.6.558. [DOI] [PubMed] [Google Scholar]
  16. Ooi C. E., Weiss J., Doerfler M. E., Elsbach P. Endotoxin-neutralizing properties of the 25 kD N-terminal fragment and a newly isolated 30 kD C-terminal fragment of the 55-60 kD bactericidal/permeability-increasing protein of human neutrophils. J Exp Med. 1991 Sep 1;174(3):649–655. doi: 10.1084/jem.174.3.649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ooi C. E., Weiss J., Elsbach P., Frangione B., Mannion B. A 25-kDa NH2-terminal fragment carries all the antibacterial activities of the human neutrophil 60-kDa bactericidal/permeability-increasing protein. J Biol Chem. 1987 Nov 5;262(31):14891–14894. [PubMed] [Google Scholar]
  18. Tobias P. S., Soldau K., Ulevitch R. J. Identification of a lipid A binding site in the acute phase reactant lipopolysaccharide binding protein. J Biol Chem. 1989 Jun 25;264(18):10867–10871. [PubMed] [Google Scholar]
  19. Tobias P. S., Ulevitch R. J. Lipopolysaccharide binding protein and CD14 in LPS dependent macrophage activation. Immunobiology. 1993 Apr;187(3-5):227–232. doi: 10.1016/S0171-2985(11)80341-4. [DOI] [PubMed] [Google Scholar]
  20. Weiss J., Beckerdite-Quagliata S., Elsbach P. Determinants of the action of phospholipases A on the envelope phospholipids of Escherichia coli. J Biol Chem. 1979 Nov 10;254(21):11010–11014. [PubMed] [Google Scholar]
  21. Weiss J., Elsbach P., Shu C., Castillo J., Grinna L., Horwitz A., Theofan G. 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. 1992 Sep;90(3):1122–1130. doi: 10.1172/JCI115930. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. White M. L., Ma J. K., Birr C. A., Trown P. W., Carroll S. F. Measurement of bactericidal/permeability-increasing protein in human body fluids by sandwich ELISA. J Immunol Methods. 1994 Jan 3;167(1-2):227–235. doi: 10.1016/0022-1759(94)90091-4. [DOI] [PubMed] [Google Scholar]
  23. Wilde C. G., Seilhamer J. J., McGrogan M., Ashton N., Snable J. L., Lane J. C., Leong S. R., Thornton M. B., Miller K. L., Scott R. W. Bactericidal/permeability-increasing protein and lipopolysaccharide (LPS)-binding protein. LPS binding properties and effects on LPS-mediated cell activation. J Biol Chem. 1994 Jul 1;269(26):17411–17416. [PubMed] [Google Scholar]
  24. Wilde C. G., Seilhamer J. J., McGrogan M., Ashton N., Snable J. L., Lane J. C., Leong S. R., Thornton M. B., Miller K. L., Scott R. W. Bactericidal/permeability-increasing protein and lipopolysaccharide (LPS)-binding protein. LPS binding properties and effects on LPS-mediated cell activation. J Biol Chem. 1994 Jul 1;269(26):17411–17416. [PubMed] [Google Scholar]

Articles from Mediators of Inflammation are provided here courtesy of Wiley

RESOURCES