Effect of anticoagulants on binding and neutralization of lipopolysaccharide by the peptide immunoglobulin conjugate CAP18(106-138)-immunoglobulin G in whole blood.

Infection and Immunity
M OgataH S Warren

Abstract

The 18-kDa cationic protein CAP18 is an antimicrobial protein isolated from rabbit granulocytes that binds lipopolysaccharide (LPS) and inhibits many of its biological activities. We covalently coupled a synthetic peptide representing amino acids 106 to 138 of CAP18 to human immunoglobulin G (IgG) by using the heterobifunctional linker N-succinimidyl-3-(2-pyridyidithio)propionate. The ability of CAP18(106-138)-IgG to bind and neutralize LPS in whole blood in the presence and absence of anticoagulants was studied. Both CAP18(106-138) and CAP18(106-138)-IgG significantly suppressed LPS-induced tumor necrosis factor (TNF) production in whole blood in the absence of anticoagulants. EDTA potentiated the ability of CAP18(106-138) and CAP18(106-138)-IgG to decrease LPS-induced TNF production in a dose-dependent manner. In contrast, heparin inhibited the ability of CAP18(106-138) and CAP18(106-138)-IgG to suppress LPS-induced TNF production. EDTA also enhanced LPS capture in a fluid-phase binding assay that utilizes magnetic anti-IgG beads to capture CAP18(106-138)-IgG (and bound [3H]LPS) in whole blood. In contrast, heparin inhibited the binding dose dependently. We conclude that CAP18(106-138)-IgG binds to and neutralizes LPS in whol...Continue Reading

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Citations

Sep 29, 2000·Critical Care Medicine·M OgataA Shigematsu
Sep 8, 2001·Canadian Journal of Anaesthesia = Journal Canadien D'anesthésie·C KawasakiA Shigematsu
Sep 4, 1999·Anesthesia and Analgesia·T KawasakiA Shigematsu
Oct 11, 2003·FEBS Letters·Herbert Bosshart, Michael Heinzelmann
Jul 15, 2003·Clinical Microbiology Reviews·Edwin S Van AmersfoortJohan Kuiper
Mar 4, 2011·Applied Microbiology and Biotechnology·Klaus BrandenburgThomas Gutsmann
Feb 9, 2010·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Ferdousi ChowdhuryAnthony Peter Williams
May 26, 1999·Infectious Disease Clinics of North America·J Hellman, H S Warren

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