HLA-antibody-induced 14C-serotonin release from platelets. A methodological analysis.
The prerequisites and kinetics of HLA-antibody-induced 14C-serotonin release from platelets were investigated with five HLA-specific antisera. By studying the influence of incubation time, temperature, pH, platelet count, stirring and extent of platelet labeling, optimal results were obtained after 30 (to 60) min of incubation at 37 degrees C, pH 7.5-8.0, under constant stirring (400 rpm). Optimal platelet counts varied between 160,000 and 400,000/microliter depending on the antiserum applied. A comparison of the amount of 14C-serotonin released by different antisera from washed platelets and from platelets in platelet-rich plasma suggests that plasma components--most likely complement--may additionally influence the results. Based on these experiments, a standardized microtechnique of HLA-antibody-induced 14C-serotonin release is recommended.
HLA-antibody-induced 14C-serotonin release from platelets. A methodological analysis. The prerequisites and kinetics of HLA-antibody-induced 14C-serotonin release from platelets were investigated with five HLA-specific antisera. By studying the influence of incubation time, temperature, pH, platelet count, stirring and extent of platelet labeling, optimal results were obtained after 30 (to 60) min of incubation at 37 degrees C, pH 7.5-8.0, under constant stirring (400 rpm). Optimal platelet counts varied between 160,000 and 400,000/microliter depending on the antiserum applied. A comparison of the amount of 14C-serotonin released by different antisera from washed platelets and from platelets in platelet-rich plasma suggests that plasma components--most likely complement--may additionally influence the results. Based on these experiments, a standardized microtechnique of HLA-antibody-induced 14C-serotonin release is recommended.