Assay of leukotriene B4 production in stimulated whole blood

We evaluated the optimal conditions for stimulating leukotriene B4 (LTB4) production in whole blood by zymosan. And the production of LTB4, LTB4 isomers and LTB4 metabolite in 19 healthy donors was compared between the stimulated whole blood method by zymosan or Ca ionophore A 23187 and the separated polymorphonuclear (PMN) method by Ca ionophore A 23187.(1) For an optimal stimulation in whole blood, EC 50 value for zymosan was 200μg/ml (whole blood), reaction time was about 20 minutes.(2) Whole blood method was characterized by lower amount of LTB4 isomers production (about 10% of the amount of LTB4 production), compared with separated PMN method.(3) The maximal amount of LTB4 produced in whole blood method by zymosan was much higher than that in separated PMN method by zymosan (from literature) .(4) There was a poor correlation between the LTB4 production (ng/106 PMN) in whole blood by zymosan or Ca ionophore A 23187 and that in separated PMN method by Ca ionophore A 23187, although there was a moderate correlation between the LTB4 production (ng/106 PMN) in whole blood method by zymosan and that in whole blood method by Ca ionophore A 23187.(5) There was almost no correlation between the LTB4 production (ng/ml) in whole blood by zymosan or Ca ionophore A 23187 and the PMN numbers in whole blood.