Development of 125I-leukotriene B4 radio-immunoassay and its clinical applications

LTB4 was extracted from porcine blood with the help of HPLC. The antiserum was raised by immunizing rabbits with LTB4 which has been conjugated with bovine serum albumin by mixed anhydride method method. The titer of the anti-body was 1:7000 and the affinity coefficient was 2.1 x 109 L/M. Its cross reactions with LTA4, LTC4, LTD4 and other arachidonic acid metabolites were 0.4 ∼ 6.4%.The 125I-histamine-LTB4 was prepared by using chloramine T procedure. The minimum detectable limit was 25 pg/tube. The recovery rate was 84.2 ∼ 113%. The intra- and inter-assay coefficient of variation were 5.2% and 9.7% respectively. The amount of LTB4 produced by normal human PMNL which were stimulated by ionophore A23187 and arachidonic acid was 695.5 ± 329.7 pg/106 cells. PMNL of patients with various acute leukemias and CML generated an increased quantities of LTB4, TXB2 and 6-keto-PGF1α. LTB4 produced by PMNL also showed an increasing tendency in both cerebral thrombosis and acute myocardial infarction, but was not altered in cerebral hemorrhage of non-infarction coronary disease