Thyroid antigens, not central tolerance, control responses to immunization in BALB/c versus C57BL/6 mice.
BACKGROUND
Vaccination with cDNA for the human thyrotropin receptor (TSHR) in a plasmid, without adjuvant, induces TSHR antibodies in C57BL/6 but rarely in BALB/c mice. This outcome could be due to a difference between "high" versus "low" antibody responder mouse strains. However, unlike their poor response to TSHR-DNA vaccination, BALB/c mice vaccinated with thyroid peroxidase (TPO)-cDNA readily develop antibodies to TPO. We hypothesized that insight into these conundrums would be provided by the following differences in central tolerance: (i) between two mouse strains (C57BL/6 versus BALB/c) for the TSHR; and (ii) between two thyroid autoantigens (TPO and the TSHR) in one mouse strain (BALB/c).


METHODS
We studied autoantigen expression using real-time polymerase chain reaction to quantify mRNA transcripts for the TSHR, TPO, and thyroglobulin (Tg) in thymic tissue (as well as in thyroid) of young mice.


RESULTS
Our hypothesis was not confirmed. Intrathymic TSHR transcript expression was similar in BALB/c and C57BL/6 mice. Moreover, thymic mRNA transcripts for TSHR and TPO were comparable. Unlike the 10-fold differences for the autoantigens in thyroid tissue (Tg greater than TPO which, in turn was greater than the TSHR), intrathymic transcripts for TPO and the TSHR were similar, both being slightly lower than the level for Tg.


CONCLUSIONS
Central tolerance, assessed by measuring intrathymic transcripts of thyroid autoantigens, does not explain the different outcome of TSHR-DNA vaccination in BALB/c and C57BL/6 mice, or even susceptibility versus resistance to hyperthyroidism induced by TSHR-adenovirus. Instead, differences in MHC and TSHR T-cell epitopes likely contribute to TSHR antibody development (or not) following DNA plasmid immunization. The greater immunogenicity of TPO versus TSHR probably relates to the greater number of nonhomologous amino acids in the human and mouse TPO ectodomains (78 amino acids) than in the human and mouse TSHR ectodomains (58 amino acids). Overall, the autoantigens themselves, not central tolerance, control DNA plasmid-induced immunity to TPO and the TSHR.
Misharin
Alexander V
AV
Autoimmune Disease Unit, Cedars-Sinai Medical Center, Los Angeles, CA, USA.
Rapoport
Basil
B
McLachlan
Sandra M
SM
D000818:Animals; D000906:Antibodies / Q000097:blood*; D001324:Autoantigens / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007108:Immune Tolerance; D007114:Immunization; D007453:Iodide Peroxidase / Q000235:genetics / Q000276:immunology*; D033862:Iron-Binding Proteins / Q000235:genetics / Q000276:immunology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D011989:Receptors, Thyrotropin / Q000235:genetics / Q000276:immunology*; D013045:Species Specificity; D013950:Thymus Gland / Q000276:immunology*; D013961:Thyroid Gland / Q000276:immunology*; D019444:Vaccines, DNA
D000906:Antibodies; D001324:Autoantigens; D033862:Iron-Binding Proteins; D012333:RNA, Messenger; D011989:Receptors, Thyrotropin; D019444:Vaccines, DNA; C056658:TPO protein, human; D007453:Iodide Peroxidase