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Two elderly patients with a bleeding tendency due to spontaneous Factor VIII inhibition were investigated. An in vitro study was designed to investigate lymphocyte transformation and the production of Factor VIII inhibitor. Blood samples from patients were separated on a ficoll-hypaque gradient and the mononuclear cells were cultured with haemophiliac plasma. Factor VIII concentrate and normal plasma as sources of Factor VIII antigen. The results showed that the patients lymphocytes were stimulated by some samples of haemophiliac plasma, but not by the Factor VIII concentrates or normal plasma. However, normal control lymphocytes were similarly stimulated by these samples of haemophiliac plasma suggesting that immune complexes were present in the haemophiliac plasma and probably caused the stimulation. Furthermore, the patients lymphocytes were stimulated with phytohaemagglutinin candida and tuberculin and normal responses were obtained with AB serum. However, the sera of both patients inhibited candida and tuberculin stimulated lymphocyte cultures and the inhibition correlated with the presence of Factor VIII inhibitor. In this study a cellular immune mechanism for the production of spontaneous Factor VIII inhibitor could not be demonstrated, however a serum inhibitor of candida and tuberculin stimulated lymphocyte cultures was found.

A Study of Lymphocyte Transformation as a Mechanism for Spontaneous Factor VIII Inhibitor Formation