Several investigators recently described a T lymphocyte subset abnormality (reversed helper/suppressor cell ratio) in haemophiliacs, especially following previous substitution therapy. We studied in vitro the potential effects of two commercial factor VIII concentrates on incorporation of 3H-thymidine by isolated human peripheral blood lymphocytes after stimulation with either phytohaemagglutinin or monoclonal antibody OKT(3). Dose-related inhibition of thymidine incorporation was found for both concentrates but none using the purified main protein constituents, i.e. factor VIII, fibrinogen or fibronectin. Furthermore, the ‘inhibitor’ was completely removed by dialysis.

Factor VIII-related properties (coagulant = Vili:C, ‘Willebrand’ factor = VIII R: WF, antigen = Vili R: AG) are measured in a constant proportion in normal plasma and certain preparations of highly purified factor VIII (relative ratios: 0.5-1.5). We tested these activities in some commercial, lyophilised concentrates of factor VIII and found a variable increase of the ratio Vili R: AG/VIII R: WF. The relative increase of VIII R:AG, and/or loss of Vili R: WF, was attributed to variable degradation of factor VIII-related protein(s) which was directly visualised by electrophoresis on 2.75% polyacrylamide gels in the presence of sodium dodecyl sulphate.

An immunoadsorbent was prepared by coupling rabbit and human antibodies against human factor VIII to Sepharose CL-2B. The resulting insoluble antibodies completely removed factor VIII and fibrinogen from normal human citrate plasma. Other coagulation factors were satisfactorily recovered in the eluted plasma. Following addition of fibrinogen, the factor VIII-deficient plasma was used for calibration of the one-stage factor VIII assay and compared with the hemophilic plasma. Parallel straight lines were obtained in the log-log plot against VIII:C, indicating that the artificial reagent can be used as a substitute for hemophilic plasma in determination of factor VIII procoagulant activity. The immunoadsorbent can be regenerated and repeatedly used for affinity chromatography binding of factor VIII.