Measurement of endotoxin activity in critically ill patients using whole blood neutrophil dependent chemiluminescence

John C Marshall, Paul M Walker, Debra M Foster, David Harris, Melanie Ribeiro, Jeff Paice, Alexander D Romaschin, Anastasia N Derzko, John C Marshall, Paul M Walker, Debra M Foster, David Harris, Melanie Ribeiro, Jeff Paice, Alexander D Romaschin, Anastasia N Derzko

Abstract

Background: Lipopolysaccharide (endotoxin) from the cell wall of Gram-negative bacteria is a potent trigger for the release of host-derived inflammatory mediators. The relationship between endotoxaemia, Gram-negative infection and the clinical syndrome of sepsis has been difficult to establish, in part because of the limitations of available endotoxin assays.

Methods: We performed an observational cohort study in critically ill patients in the medical/surgical intensive care unit (ICU) of a tertiary care hospital. Whole blood endotoxin levels on the day of ICU admission were measured using a novel chemiluminescent assay--the endotoxin activity assay (EAA)--and the chromogenic modification of the limulus amoebocyte lysate (LAL) assay.

Results: We studied 74 consecutive admissions. Endotoxin levels were higher in patients with a diagnosis of sepsis (470 +/- 57 pg/ml) than in patients admitted with a diagnosis other than sepsis (157 +/- 140 pg/ml; P < 0.001). Endotoxaemia was significantly associated with Gram-negative infection (P < 0.05); no patient with a Gram-negative infection had an endotoxin level below 50 pg/ml. White blood cell counts of patients with EAA-detected endotoxaemia were significantly higher (15.7 +/- 9.1 x 10(9) cells/l for endotoxaemic patients versus 10.8 +/- 6.2 x 10(9) cells/l for patients without endotoxaemia; P < 0.05).

Conclusion: Endotoxaemia is associated with Gram-negative infection from any source, and with a diagnosis of sepsis and leukocytosis. These correlations were not apparent using the LAL method. The EAA may be a useful diagnostic tool for the investigation of invasive Gram-negative infection and incipient sepsis.

Figures

Figure 1
Figure 1
Effect of polymyxin-sulphate preincubation on endotoxin recovery in the endotoxin activity assay. Samples from 15 intensive care unit patients with endotoxaemia ranging from 50 to more than 800 pg/ml were pretreated with polymyxin-sulphate at a concentration of 300 g/ml for 20 min at 37°C. Endotoxin concentrations are expressed as means ± standard deviation.
Figure 2
Figure 2
Endotoxin levels in patients with Gram-negative infection. Endotoxin levels were significantly higher by endotoxin activity assay (P < 0.05, n = 10). Endotoxin concentrations are expressed as means ± standard deviation.

References

    1. Morrison DC, Ulevitch RJ. The effects of bacterial endotoxin on host mediation systems. Am J Pathol. 1978;93:527–601.
    1. Parillo JE. Pathogenic mechanisms of septic shock. N Engl J Med. 1993;328:1471–1477.
    1. Watson RWG, Redmond HP, Bouchier-Hayes D. Role of endotoxin in mononuclear phagocyte-mediated inflammatory responses. J Leukoc Biol. 1994;56:95–103.
    1. Santos AA, Wilmore DW. The systemic inflammatory response: perspective of human endotoxemia. Shock. 1996;6(suppl 1):S50–S56.
    1. Danner RL, Elin RJ, Hosseini JM, Wesley RA, Reilly JM, Parilllo JE. Endotoxemia in human septic shock. Chest. 1991;99:169–175.
    1. Casey LC, Balk RA, Bone RC. Plasma cytokine and endotoxin levels correlate with survival in patients with the sepsis syndrome. Ann Intern Med. 1993;119:771–778.
    1. Wortel CH, von der Mohlen MA, van Deventer SJ, Sprung CL, Jastremski M, Lubbers MJ, Smith CR, Allen IE, ten Cate JW. Effectiveness of a human monoclonal anti-endotoxin antibody (HA-1A) in Gram negative sepsis: relationship to endotoxin and cytokine levels. J Infect Dis. 1992;166:1367–1374.
    1. Bion JF, Badger I, Crosby HA, Hutchings P, Kong KL, Baker J, Hutton P, McMaster P, Buckels JA, Elliott TS. Selective decontamination of the digestive tract reduces Gram-negative pulmonary colonization but not systemic endotoxemia in patients undergoing elective liver transplantation. Crit Care Med. 1994;22:40–49.
    1. Gaeta GB, Perna P, Adinolfi LE, Utili R, Ruggiero G. Endotoxemia in a series of 104 patients with chronic liver diseases: prevalence and significance. Digestion. 1982;23:239–244.
    1. Roumen RMH, Frieling JTM, van Tits HWHJ, van der Vliet JA, Goris RJ. Endotoxemia after major vascular operations. J Vasc Surg. 1993;18:853–857.
    1. Schlag G, Redl H, Dinges HP, Davies J. Sources of endotoxin in the posttraumatic setting. In: Sturk A, van Deventer SJH, ten Cate JW, Buller HR, Thijs LG, Levin J, editor. In Bacterial Endotoxins: Cytokine Mediators and New Therapies for Sepsis. Amsterdam: Wiley-Liss Inc.; 1991. pp. 121–134.
    1. Jansen PG, Te Velthuis H, Oudemans-Van Straaten HM, Bulder ER, Van Deventer SJ, Sturk A, Eijsman L, Wildevuur CR. Perfusion-related factors of endotoxin release during cardiopulmonary bypass. Eur J Cardiothorac Surg. 1994;8:125–129.
    1. Natanson C, Hoffman WD, Suffredini AF, Eichacker PQ, Danner RL. Selected treatment strategies for septic shock based on proposed mechanisms of pathogenisis. Ann Intern Med. 1994;120:771–783.
    1. Rangel-Frausto MS, Pittet D, Costigan M, Hwang T, Davis CS, Wenzel RP. The natural history of the systemic inflammatory response syndrome (SIRS). A prospective study. JAMA. 1995;272:117–123.
    1. Ziegler EJ, Fisher CJ, Jr, Sprung CL, Straube RC, Sadoff JC, Foulke GE, Wortel CH, Fink MP, Dellinger RP, Teng NN, et al. Treatment of Gram-negative bacteremia and septic shock with HA-1A human monoclonal antibody against endotoxin. N Engl J Med. 1991;324:429–436.
    1. van Deventer SJH, Buller HR, ten Cate JW, Sturk A, Pauw W. Endotoxemia: an early predictor of septicemia in febrile patients. Lancet. 1988;1:605–608.
    1. Hurley JC. Antibiotic-induced release of endotoxin: a reappraisal. Clin Infect Dis. 1992;15:840–854.
    1. Elin RJ, Robinson RA, Levine AS, Wolff SM. Lack of clinical usefulness of the limulus test in the diagnosis of endotoxemia. N Engl J Med. 1975;293:521–524.
    1. Levin J, Bang FB. Clottable protein in limulus: its localization and kinetics of its coagulation by endotoxin. Thromb Diath Haemorrh. 1968;19:186–197.
    1. Tamura H, Tanaka S, Obayashi T, Yoshida M, Kawai T. A new sensitive method for determining endotoxin in whole blood. Clin Chim Acta. 1991;200:35–42.
    1. Bates DW, Parsonnet J, Ketchum PA, Novitsky TJ, Sands K, Hibberd PL, Graman PS, Lanken PN. Limulus amebocyte lysate assay for detection of endotoxin in patients with sepsis syndrome. AMCC Sepsis Project Working Group. Clin Infect Dis. 1998;27:582–591.
    1. Romaschin AD, Harris DM, Ribeiro MB, Paice J, Foster DM, Walker PM, Marshall JC. A rapid assay of endotoxin in whole blood using autologous neutrophil dependent chemiluminescence. J Immunol Methods. 1998;212:169–185.
    1. Marshall JC, Cook DJ, Christou NV, Bernard GR, Sprung CL, Sibbald WJ. Multiple organ dysfunction score: a reliable descriptor of a complex clinical outcome. Crit Care Med. 1995;23:1638–1652.
    1. Knaus WA, Draper EA, Wagner DP, Zimmerman JE. APACHE II: a severity of disease classification system. Crit Care Med. 1985;13:818–829.
    1. Members of the American College of Chest Physicians/Society of Critical Care Medicine Consensus Conference Definitions of sepsis and organ failure and guidelines for the use of innovative therapies in sepsis. Crit Care Med. 1992;23:376–393.
    1. Hurley JC. Re-appraisal with meta analysis of bacteremia, endotoxemia and mortality in Gram negative sepsis. J Clin Microbiol. 1995;33:1278–1282.
    1. Zhang GH, Baek L, Bertelen T, Koch C. Quantification of the endotoxin-neutralizing capacity of serum and plasma. APMIS. 1995;103:712–730.
    1. Behre G, Schedel I, Nentwig B, Wormann B, Essink M, Hiddemann W. Endotoxin concentration in neutropenic patients with suspected Gram-negative sepsis: correlation with clinical outcome and determination of anti-endotoxin core antibodies during therapy with polyclonal immunoglobulin M-enriched immunoglobulins. Antimicrob Agents Chemother. 1992;36:2139–2146.
    1. Brandtzaeg P, Kierulf P, Gaustad P, Skulberg A, Bruun JN, Halvorsen S, Sorensen E. Plasma endotoxin as a predictor of multiple organ failure and death in systemic meningococcal disease. J Infect Dis. 1989;159:195–204.
    1. Exley AR, Leese T, Holliday MP, Swann RA, Cohen J. Endotoxemia and serum tumour necrosis factor as prognostic markers in severe acute pancreatitis. Gut. 1992;33:1126–1128.
    1. Yao YM, Sheng ZY, Tian HM, Wang YP, Guo ZR, Gao WY. The association of circulating endotoxemia with the development of multiple organ failure in burned patients. Burns. 1995;21:255–258.
    1. Go LL, Healey PJ, Watkins SC, Simmons RL, ROWE MI. The effect of endotoxin on intestinal mucosal permeability to bacteria in vitro. Arch Surg. 1995;130:53–58.
    1. Shenep JL, Flynn PM, Barrett FF, Stidham GL, Westenkirchner DF. Serial quantitation of endotoxemia and bacteremia during therapy for Gram-negative bacterial sepsis. J Infect Dis. 1988;157:565–568.
    1. Nahum A, Hoyt J, Schmitz L, Moody J, Shapiro R, Marini JJ. Effect of mechanical ventilation strategy on dissemination of intra-tracheally instilled Escherichia coli in dogs. Crit Care Med. 1997;25:1733–1743.
    1. Opal SM, Scannon PJ, Vincent JL, White M, Carroll SF, Palardy JE, Parejo NA, Pribble JP, Lemke JH. Relationship between plasma levels of lipopolysaccharide (LPS) and LPS-binding protein in patients with severe sepsis and septic shock. J Infect Dis. 1999;180:1584–1589.

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