Figure 1.
A, SDS-PAGE comparison of recombinant TSHR A-subunit protein generated in CHO cells and bacteria as a fusion protein with Trx. The bacterial fusion partner (Trx) is included. B, Schedule for pretreatment before immunization with A-subunit adenovirus. BALB/c mice were injected with A-sub-CHO or controls 1 wk before the first of two A-sub-Ad immunizations. Controls include saline (0) or other proteins. A separate group of mice was immunized with Con-Ad. One week after the second immunization, sera were obtained for analysis. C, T4 levels (micrograms per deciliter) for individual mice (five per group). The shaded area represents the mean ± 2 sd levels in Con-Ad immunized mice. *, Significantly reduced vs. pretreatment without A-sub-CHO (0 μg); t test, P = 0.009. D, TSAb activity for the mice shown in C. The data were obtained using mouse TSHR-expressing CHO cells; similar observations were made with human TSHR-expressing CHO cells (not shown). TSAb values are presented as the mean + sem for cAMP release expressed as a percentage of the response from Con-Ad immunized mice. *, Significantly reduced vs. pretreatment with 0 μg A-sub-CHO (rank sum test, P = 0.032).