Female mice of the following strains (5C8 wk of age; Jackson Laboratory, Bar Harbor, ME) were studied: These mice are subsequently referred to as C3H, B6, and F1

Female mice of the following strains (5C8 wk of age; Jackson Laboratory, Bar Harbor, ME) were studied: These mice are subsequently referred to as C3H, B6, and F1. the susceptibility to induced TSHR antibodies hyperthyroidism. Importantly, BXH and CXB mice share genetic loci controlling the generation of TSHR antibodies (Chr 17, major histocompatibility complex region, and Chr X) and development of hyperthyroidism (Chr 1 and 3). Moreover, some chromosomal linkages are unique to either BXH or CXB strains. An interesting candidate gene linked to thyroid-stimulating Budesonide antibody generation in BXH mice is the Ig heavy chain locus, suggesting a role for particular germline region genes as precursors for these antibodies. In conclusion, our findings reinforce the importance of major histocompatibility complex region genes in controlling Budesonide the generation of TSHR antibodies measured by TSH binding inhibition. Moreover, these data emphasize the value of RI strains to dissect the genetic basis for MAP2 induced TSHR antibodies their effects on thyroid function in Graves disease. THYROID-STIMULATING antibodies (TSAb) and hyperthyroidism characteristic of Graves disease can be induced in some, but not all, mouse strains using immunization approaches involving expression of the TSH receptor (TSHR). For example, mice of the BALB/c strain are susceptible to hyperthyroidism induced by injecting TSHR-expressing B cells (1) or dendritic cells (2) and by immunization with adenovirus encoding the TSHR or its A-subunit (3,4). On the other hand, C57BL/6 (B6) strain mice immunized with TSHR- or A-subunit adenovirus Budesonide develop TSHR antibodies but not hyperthyroidism (3,5). CBA/J mice immunized with either TSHR-expressing fibroblasts (6) or TSHR-adenovirus (3) have poor TSHR antibody responses and do not become hyperthyroid. Non-major histocompatibility complex (non-MHC) genes contribute to this divergence of responses. For example, BALB/k and BALB/c strains, with different MHC but the same background genes, become hyperthyroid after TSHR-adenovirus immunization (4,7). Studies on other mouse strains immunized with TSHR-fibroblasts or TSHR-adenovirus provide support for the involvement of non-MHC genes in the development of hyperthyroidism (6,7,8) (reviewed in Ref. 9). More detailed information on the genetic susceptibility to A-subunit adenovirus immunization has recently been obtained using recombinant inbred (RI) strains of mice whose genomes have been characterized (10). The first whole genome scan in a murine model of Graves disease was performed using RI strains derived by repeated brother sister matings of the progeny of BALB/c and B6 parents (hence termed CXB mice). The outcome of TSHR A-subunit adenovirus immunization of CXB strains pointed to several loci on different chromosomes (Chr) that separately controlled TSHR antibody generation and hyperthyroidism (11). High-resolution genetic maps have been generated for four other sets of RI strains that, like CXB, share B6 as one of the parental strains (10,12). One of these RI strains is designated BXH because it is derived from crosses of B6 and C3H/He parents. The C3H/He parental strain is known to develop Graves hyperthyroidism after TSHR-fibroblast injection (6), but its response to TSHR-adenovirus immunization has not been determined. In the present study, we tested C3H/He mice and the F1 offspring of this strain crossed to B6 mice for their susceptibility to hyperthyroidism induced by immunization with TSHR A-subunit adenovirus. In addition, we investigated the responses of 13 BXH strains immunized in the same way. Our data confirm the findings in CXB mice (11) that genes on multiple Chr contribute separately to the variations in TSHR antibody generation and the induction of hyperthyroidism. Moreover, the observations Budesonide from the previous (CXB) and present (BXH) studies provide evidence for shared as well as unique genetic loci that influence the induction of TSHR antibodies and hyperthyroidism in the TSHR-adenovirus mouse model of Graves disease. Materials and Methods Immunization of mice with TSHR A-subunit adenovirus Adenovirus expressing the human TSHR A-subunit (A-subunit-Ad, amino acid residues 1C289) and Budesonide null adenovirus [control adenovirus (Con-Ad)] have been described (4,13). Adenoviruses were propagated in HEK293 cells (American Type.