Open AccessNew approach in flow‐cytometric determination of endotoxin during endotoxic shock
Author(s) -
Staubach K.H.,
Nolde J.,
Brade H.,
Woltmann A.,
Bruch H.P.
Publication year - 2000
Publication title -
british journal of surgery
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.202
H-Index - 201
eISSN - 1365-2168
pISSN - 0007-1323
DOI - 10.1046/j.1365-2168.2000.01544-63.x
Subject(s) - peripheral blood mononuclear cell , medicine , limulus , lipopolysaccharide , flow cytometry , monoclonal antibody , shock (circulatory) , septic shock , immunology , lysis , andrology , antibody , pharmacology , in vitro , sepsis , biology , biochemistry , paleontology
Background Serum endotoxin was formerly measured with the non‐specific Limulus lysate assay. The present approach was to quantitate the amount of endotoxin bound by peripheral mononuclear cells in order to develop a method for the diagnosis of early septic shock. Methods Using a murine monoclonal antibody (WN1‐222/5), which binds highly specifically to lipopolysaccharide (LPS), a new method for measuring the amount of LPS bound to peripheral mononuclear cells was developed. Ten pigs were studied under sedation and peripheral mononuclear cells were taken every 4 h to determine the concentration of endotoxin by flow cytometry. The results are shown in the Table. ResultsTime after LPS infusion (h) 0 1 4 6 8Marked cells (%) 32 61 75 72 85The percentage of marked mononuclear cells increased during shock. Only in the last hours before death did the rate of increase decline. Conclusion Preliminary data on marked mononuclear cells showed that the amount of natural incorporated endotoxin, i.e. the quantity of bound endotoxin before infusion, was 32 per cent. © 2000 British Journal of Surgery Society Ltd