title: Mechanisms of thrombin generation during surgery and cardiopulmonary bypass.
abstract: Although in vitro studies have been invaluable in revealing the complex biochemistry of the blood coagulation system, the mechanisms involved during the in vivo response to hypercoagulable stimuli are still unclear. We have used plasma-based enzyme-linked immunosorbent assays (ELISAs) to study the mechanisms by which the coagulation system is activated in vivo during human cardiopulmonary bypass (CPB) surgery (n = 8). A novel immunoassay for factor XIIa was used to detect activation of the contact system, factor IX activation peptide (FIXAP) was used as a marker for activation of factor IX, and prothrombin fragment F1 + 2 (F1 + 2) was used as a marker for thrombin generation. The ELISA for FIXAP is described for the first time herein. F1 + 2 levels increased early in response to surgical intervention: from a baseline of 38.7 +/- 9.7 ng/mL (mean +/-SE), levels increased rapidly during surgery and bypass to a maximum of 448.5 +/- 92.0 ng/mL. A modest yet significant increase in factor XIIa levels from 3.47 +/- 0.54 ng/mL to 4.33 +/- 0.85 ng/mL was evident during surgery before bypass, but no further significant increase was detected on establishing extracorporeal circulation. FIXAP levels demonstrated a small and late increase during surgery from 4.98 +/- 0.55 ng/mL to a maximum of 10.20 +/- 1.23 ng/mL, the increase beginning at the time of near maximal F1 + 2 levels. There was no association between activation of the contact system (factor XIIa levels) and the generation of thrombin (F1 + 2 levels). However, a strong association (r = .705) was apparent between the generation of thrombin (F1 + 2 levels) and activation of factor IX (FIXAP levels), despite the delay between the activation of prothrombin and factor IX. The data do not support the established view that contact activation resulting from exposure of blood to foreign surfaces is the major procoagulant stimulus in CPB. Instead, the results suggest that the main trigger to coagulation during CPB surgery was provided via the tissue factor-factor VIIa mechanism in response to the cutting of blood vessels, which directly activated factor X and then prothrombin. The late activation of factor IX, which presumably also contributed to maximal prothrombin activation, could have arisen due to direct tissue factor-factor VIIa action, or by secondary feedback action of thrombin on the intrinsic system.
authors.affiliation: Department of Haematology, Charing Cross and Westminster Medical School, London, UK.
authors.forename: M D
authors.initials: MD
authors.lastname: Boisclair
authors.forename: D A
authors.initials: DA
authors.lastname: Lane
authors.forename: H
authors.initials: H
authors.lastname: Philippou
authors.forename: M P
authors.initials: MP
authors.lastname: Esnouf
authors.forename: S
authors.initials: S
authors.lastname: Sheikh
authors.forename: B
authors.initials: B
authors.lastname: Hunt
authors.forename: K J
authors.initials: KJ
authors.lastname: Smith
mesh_terms: D000368:Aged; D000595:Amino Acid Sequence; D002315:Cardiopulmonary Bypass; D004797:Enzyme-Linked Immunosorbent Assay; D005172:Factor XI / Q000378:metabolism; D015956:Factor XIIa / Q000032:analysis; D006801:Humans; D008875:Middle Aged; D008969:Molecular Sequence Data; D011516:Prothrombin / Q000378:metabolism; D013917:Thrombin / Q000096:biosynthesis*
chemical_list: D011516:Prothrombin; D005172:Factor XI; D015956:Factor XIIa; D013917:Thrombin