Dose-response relationships in a cytochemical section bioassay for thyroid stimulators

The thyroid stimulators TSH and thyroid stimulating immunoglobulins (TSI) have previously been distinguished in a cytochemical section bioassay (CBA) by the different times at which they caused maximum increase of lysosomal membrane permeability within guinea-pig thyroid follicular cells. The present study demonstrates that the times at which maximum stimulation occurs in the assay depends not only on the type (TSH or TSI) but also on the concentration of the stimulator. At higher concentrations of stimulator earlier times of maximum stimulation were observed for both TSH and TSI. When both stimulators were simultaneously present in plasma at higher concentrations a single peak of stimulatory activity occurred. Neutralization studies using specific antisera revealed that the single peak represented a merging of TSH and TSI peaks. These findings explain the lack of parallelism to the standard curve seen at higher concentrations of TSH or TSI in the CBA. They emphasize the necessity of measuring plasma samples at a variety of dilutions in order to achieve parallelism and to avoid intra-assay interference between TSH and TSI. When these conditions are observed CBA can be used to detect TSH and TSI with extreme sensitivity even when both are simultaneously present in plasma.