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  • Reduced effectiveness of CD4+Foxp3+ regulatory T cells in CD28-deficient NOD.H-2h4 mice leads to increased severity of spontaneous autoimmune thyroiditis.

Reduced effectiveness of CD4+Foxp3+ regulatory T cells in CD28-deficient NOD.H-2h4 mice leads to increased severity of spontaneous autoimmune thyroiditis.

Journal of immunology (Baltimore, Md. : 1950) (2013-10-08)
Jason S Ellis, So-Hee Hong, Habib Zaghouani, Helen Braley-Mullen
ABSTRACT

NOD.H-2h4 mice given NaI in their drinking water develop iodine-accelerated spontaneous autoimmune thyroiditis (ISAT) with chronic inflammation of the thyroid by T and B cells and production of anti-mouse thyroglobulin (MTg) autoantibody. CD28(-/-) NOD.H-2h4 mice, which have reduced numbers of CD4(+)Foxp3(+) regulatory T cells (Tregs), were developed to examine the role of Tregs in ISAT development. CD28(-/-) NOD.H2-h4 mice develop more severe ISAT than do wild-type (WT) mice, with collagen deposition (fibrosis) and low serum T4. CD28(-/-) mice have increased expression of proinflammatory cytokines IFN-γ and IL-6, consistent with increased mononuclear cell infiltration and tissue destruction in thyroids. Importantly, transferring purified CD4(+)Foxp3(+) Tregs from WT mice reduces ISAT severity in CD28(-/-) mice without increasing the total number of Tregs, suggesting that endogenous Tregs in CD28(-/-) mice are functionally ineffective. Endogenous CD28(-/-) Tregs have reduced surface expression of CD27, TNFR2 p75, and glucocorticoid-induced TNFR-related protein compared with transferred CD28(+/+) Tregs. Although anti-MTg autoantibody levels generally correlate with ISAT severity scores in WT mice, CD28(-/-) mice have lower anti-MTg autoantibody responses than do WT mice. The percentages of follicular B cells are decreased and those of marginal zone B cells are increased in spleens of CD28(-/-) mice, and they have fewer thyroid-infiltrating B cells than do WT mice. This suggests that CD28 deficiency has direct and indirect effects on the B cell compartment. B cell-deficient (B(-/-)) NOD.H-2h4 mice are resistant to ISAT, but CD28(-/-)B(-/-) mice develop ISAT comparable to WT mice and have reduced numbers of Tregs compared with WT B(-/-) mice.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
L-Thyroxine, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
L-Thyroxine, ≥98% (HPLC)
Sigma-Aldrich
Thyroglobulin from porcine thyroid gland, powder
Sigma-Aldrich
Sodium iodide, AnhydroBeads, −10 mesh, 99.999% trace metals basis
Sigma-Aldrich
Sodium iodide, ≥99.99% trace metals basis
Sigma-Aldrich
Sodium iodide, ACS reagent, ≥99.5%
Sigma-Aldrich
Sodium iodide, ReagentPlus®, ≥99%
Sigma-Aldrich
Thyroglobulin human
Sigma-Aldrich
Sodium iodide, 99.999% trace metals basis
Supelco
L-Thyroxine (T4) solution, 100 μg/mL in methanol with 0.1N NH3, ampule of 1 mL, certified reference material, Cerilliant®
Sigma-Aldrich
Sodium iodide, puriss., meets analytical specification of Ph. Eur., BP, USP, 99-100.5% (calc. to the dried substance)