Using a combined pretreatment methodology employing an alkaline reagent and a kinetic chromogenic Limulus test using Endospecy®, different types of endotoxins in murine plasma were able to be measured quantitatively. The detection of smooth (S)-form Salmonella abortus equi LPS (S-LPS) was approximately twice as sensitive as Helicobacter pylori LPS in aqueous solution. S-LPS added to whole mouse blood was recovered quantitatively from plasma, but the rough (R)-form S. minnesota LPS was not. The rates of clearance of S-LPS from the circulation following intravenous injection were shown to be different among BALB/c, C3H/HeN and B10Sn mice. Infection of mice with Salmonella typhimurium 3 days earlier resulted in accelerated clearance of subsequently injected LPS from the circulation. Significantly higher amounts of free non-microbe-associated LPS were detected in the blood of mice 1 h after infection with Escherichia coli O111:B4 followed by immediate treatment with ceftazidime, in comparison with non-antibiotic-treated but infected mice.
Get full access to this article
View all access options for this article.
References
1.
Levin J., Bang F.B.Clotable protein in Limulus: its localization and kinetics of its coagulation by endotoxin. Thromb Diath Haemorrh1968; 19: 186-197.
2.
Iwanaga S., Morita T., Harada T. et al. Chromogenic substrates for horseshoe crab clotting enzyme: its application for the assay of bacterial endotoxins. Haemostasis1978; 7: 183-188.
3.
Tanaka S., Iwanaga S.Limulus test for detecting bacterial endotoxins. Methods Enzymol1993; 223: 358-364.
4.
Hurley J.C.Endotoxemia: methods of detection and clinical correlates. Clin Microbiol Rev1995; 8: 268-292.
5.
Tamura H., Arimoto Y., Tanaka S., Yoshida M., Obayashi T., Kawai T.Automated kinetic assay for endotoxin and (1→3)-β-D-glucan in human blood. Clin Chim Acta1994; 226: 109-112.
6.
Tamura H., Tanaka S., Obayashi T., Yoshida M., Kawai T.A new sensitive microplate assay of plasma endotoidn. J Clin Lab Anal1992; 6: 232-238.
7.
Galanos C., Luderitz O., Westphal O.A new method for the extraction of R lipopolysaccharides. Eur J Biochem1969; 9: 245-249.
8.
Galanos C., Lüderitz O., Westphal O.Preparation and properties of a standardized lipopolysaccharide from Salmonella abortus equi (Novo-Pyrexal). Zbl Bakt Hyg I Abt Orig A1979; 243: 226-244.
9.
Takakuwa T., Knopf H.P., Sing A., Carsetti R., Galanos C., Freudenberg M.A.Induction of CD14 expression in Lpsn, Lpsd and tumor necrosis factor receptor-deficient mice. Eur J Immunol1996 ; 26: 2686-2692.
10.
Galanos C., Freudenberg M.A., Reutter W.Galactosamine-induced sensitization to the lethal effects of endotoxin . Proc Natl Acad Sci USA1979; 76: 5939-5943.
11.
Kumazawa Y. , Freudenberg M.A., Hausmann C., Meding-Slade S., Langhorne J., Galanos C.Formation of interferon-gamma and tumor necrosis factor in mice during Salmonella typhimurium infection. Pathobiology1991; 59: 194-196.
12.
Freudenberg M.A., Kumazawa Y., Meding S., Langhorne J., Galanos C.Gamma interferon production in endotoxin-responder and -nonresponder mice during infection . Infect Immun1991 ; 59: 3484-3491.
13.
Bucklin S.E. , Morrison D.C.Differences in therapeutic efficacy among cell wall-active antibiotics in a mouse model of Gram-negative sepsis . J Infect Dis1995; 172: 1519-1527.
14.
Morita H.,Urashima Y., Tanaka S., Aketagawa J., Tamura H., Kumazawa Y.Simultaneous estimation of endotoxin and (1→3)-β-D-glucan with an automated kinetic-chromogenic assay using Wellreader. J Antibact Antifung Agents1996; 24: 467-475.
15.
Obayashi T.Addition of perchloric acid to blood samples for colorimetric Limulus test using chromogenic substrate: comparison with conventional procedures and clinical applications. J Lab Clin Med1984; 104: 321-330.
16.
Obayashi T. , Tamura H., Tanaka S., Ohki M., Takahashi S., Kawai T.Endotoxin-inactivating activity in normal and pathological human blood samples. Infect Immun1986; 53: 294-292
17.
Fomsgaard A. , Freudenberg M.A., Galanos C.Modification of the silver staining technique to detect lipopolysaccharide in polyacrylamide gels . J Clin Microbiol1990; 28: 2627-2631.
18.
Tsai C-M., Frasch C.A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem1982; 119: 115-119.
19.
Musson R.A. , Morrison D.C., Ulevitch R.J.Distribution of endotoxin (lipopolysaccharide) in the tissues of lipopolysaccharide-responsive and -unresponsive mice. Infect Immun1978; 21: 448-457.
20.
Jackson J.J. , Kropp H.β-Lactam antibiotic-induced release of free endotoxin: in vitro comparison of penicillin-binding protein (PBP) 2-specific imipenem and PBP 3-specific ceftazidime. J Infect Dis1992; 165: 1033-1041.
21.
Neely A.N., Holder I.A.A murine model with aspects of clinical relevance for the study of antibiotic-induced endotoxin release in septic hosts. j Endotoxin Res1996; 3: 229-235.
22.
Morrison D.C. , Bucklin S.E., Leeson M.C., Norimatsu M.Contribution of soluble endotoxin released from Gram-negative bacteria by antibiotics to the pathogenesis of experimental sepsis in mice. J Endotoxin Res1996; 3: 237-243.
23.
Opal S.M., Horn D.L., Palardy J.E. et al. The in vivo significance of antibiotic-induced endotoxin release in experimental Gram-negative sepsis. J Endotoxin Res1996; 3: 245-252.
24.
Galanos C., Lüderitz O., Rietschel E.T. et al. Synthetic and natural Escherichia coli free lipid A express identical endotoxic activities. Eur J Biochem1985; 148: 1-5.
25.
Kotani S., Takada H., Tsujimoto M. et al. Synthetic lipid A with endotoxic and related biological activities comparable to those of natural lipid A from an Escherichia coli Re-mutant . Infect Immun1985; 49: 225-237.
26.
Homma J.Y., Matsuura M., Kanegasaki S. et al. Structural requirements of lipid A responsible for the functions: a study with chemically synthesized lipid A and its analogues. J Biochem (Tokyo) 1985; 98: 395-406.
27.
Carr Jr C., Morrison D.C.Lipopolysaccharide interaction with rabbit erythrocyte membrane. Infect Immun1984; 43: 600-606.
28.
Carr Jr C., Morrison D.C.Mechanism of polymyxin B-mediated lysis of lipopolysaccharide-treated erythrocytes . Infect Immun1985; 49: 84-89.
29.
Kirikae T., Inada K., Hirata M., Yoshida M., Kondo S., Hisatsune K.Identification of Re lipopolysaccharide-binding protein on murine erythrocyte membrane. Microbiol Immunol1988 ; 32: 33-44.
30.
Preston A., Mandrell R.E., Gibson B.W., Apicella M.A.The lipooligosaccharides of pathogenic gram-negative bacteria. Crit. Rev. Microbiol.1996; 22: 139-130.
31.
Freudenberg M.A. , Kleine B., Freudenberg N., Galanos C.Distribution, degradation and elimination of intravenously applied lipopolysaccharide in rats. In: Homma J.Y., Kanegasaki S. , Lüderitz O., Shiba T., Westphal O. (eds) Bacterial Endotoxin- Chemical, Biological Clinical Aspects. Weinheim: Verlag Chemie, 1984; 295-304.
32.
Freudenberg M.A. , Galanos C.Metabolism of LPS in vivo. In: Morrison D.C., Ryan J.L. (eds) Bacterial Endotoxic Lipopolysaccharides , Vol. 2. Boca Raton: CRC Press, 1992; 275-294.
