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Measurement of different types of endotoxin in murine plasma by pretreatment with alkaline reagent and kinetic chromogenic Limulus test

Seikagaku Corp., Tokyo, Japan

R. Hasunuma

Department of Biosciences, School of Science, Kitasato University, Sagamihara, Japan

M. Fujihara

Department of Biochemistry, School of Allied Medical Sciences, Kitasato University, Sagamihara, Japan

S. Tanaka

Seikagaku Corp., Tokyo, Japan

C. Galanos

Max Planck Institut für Immunbiologie, Freiburg, Germany

M.A. Freudenberg

Max Planck Institut für Immunbiologie, Freiburg, Germany

D.C. Morrison

University of Kansas Medical Center, Kansas City, Kansas, USA

Y. Kumazawa

Department of Biosciences, School of Science, Kitasato University, Sagamihara, Japan, kumazawa{at}jet.sci.kitasato-u.ac.jp

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.

Journal of Endotoxin Research, Vol. 4, No. 5, 339-346 (1997)
DOI: 10.1177/096805199700400505


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