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THE INDIRECT LEUCOCYTE MIGRATION INHIBITION ASSAY — AN ENDOTOXIN‐SENSITIVE CHEMOKINETIC ASSAY. PART II
Author(s) -
Remvig L.,
Baek L.,
Kristensen E.
Publication year - 1986
Publication title -
acta pathologica microbiologica scandinavica series c: immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.909
H-Index - 88
eISSN - 1600-0463
pISSN - 0108-0202
DOI - 10.1111/j.1699-0463.1986.tb02090.x
Subject(s) - lipopolysaccharide , chemokinesis , agarose , chemistry , microbiology and biotechnology , microgram , immunology , pharmacology , andrology , in vitro , chemotaxis , medicine , chromatography , biochemistry , biology , receptor
Depletion of agarose for endotoxins resulted in a low spontaneous migration of polymorphnuclear cells (PMNC). Re‐addition of endotoxin, in casu lipopolysaccharide from E. coli 026:B6 (LPS), enhanced the spontaneous PMNC migration in a two‐phased dose‐response pattern, reaching maximum migration with LPS 1 times 10 −7 g/ml. Thus, the migration of PMNC under agarose seems to be a chemokinesis. Leucocyte migration inhibition factor (LIF), induced by PPD 50 μg/ml at endotoxin‐free conditions, significantly reduced the PMNC migration compared to supernatants from control cultures, however not compared to the conventional limit of significance, MI = 0.80. With increasing PMNC migration there was an insignificant decrease in the MI. Addition of LPS, 1 times 10 −9 g/ml, during LIF induction caused a significant increase in LIF production, an effect which overshadowed the effect of PPD. Thus, the application of the conventional limit of significance, MI = 0.80, may result in false‐negative or false‐positive conclusions, depending upon the endotoxin contamination. A standardization of the endotoxin content in both steps of the indirect leucocyte migration inhibition assay seems mandatory in order to obtain a reliable and reproducible bioassay.