Cobra venom factor was used for the detection of factor B synthesized by mouse peritoneal macrophages. This method was shown to be specific for factor B assay by neutralization by antimouse factor B antibody. The amount of factor B in the culture supernatant, assessed by this method, was found to be dependent on the medium used for cultivation of macrophages. The addition of 25% L cell‐conditioned medium to minimal essential medium (LCM‐MEM) enhanced the production of factor B and also of lysozyme. Kinetic analysis in LCM‐MEM showed that factor B produced by 6 × 104 cells/cm2 increased up to 72 hr and reached a plateau at 96 hr. The amounts of factor B and lysozyme produced in LCM‐MEM depended upon the number of macrophages. Production of factor B was completely inhibited by 1 μg of cycloheximide per ml and was restored by its removal.
SummaryThe nature of sugar chain of factor VTII/von Willebrand factor in plasma of normal subjects and patients with von Willebrand’s disease (vWd) was examined by crossed affinoimmunoelectrophoresis using anti-human factor VIII rabbit serum, with inserted Ricinus communis agglutinin-120 (RCA-120) agarose layer (RCA – CIE). Molecular weights of factor VlU-related antigen (VIIIR: Ag) were estimated by SDS polyacrylamide gel electrophoresis — crossed affinoimmunoelectrophoresis (SDS PAGE – RCA – CIE).VIIIR :Ag, in normal plasma and in classical form of vWd, showed two precipitin peaks on RCA – CIE. The slower moving component of VIIIR :Ag with molecular weights over 3×106 daltons from normal subjects and patients with classical form of vWd showed a high affinity for RCA-120. The faster moving component of VIIIR: Ag below 3×106 daltons from the abovementioned subjects and patients with a variant form (Type IIA) showed a very weak affinity for RCA-120.These results suggested that all of VIIIR: Ag in these variant cases may have a deficiency of galactose residues reactive with RCA, in addition to an incomplete polymerization of VIIIR: Ag, similar to that of the faster moving component of normal subjects.
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