CXCR4 antagonist AMD3100 ameliorates thyroid damage in autoimmune thyroiditis in NOD.H­2h4 mice.

CXC chemokine ligand 12 (CXCL12) and its receptor, CXC chemokine receptor 4 (CXCR4), are upregulated in mice with autoimmune thyroid diseases. However, whether this interaction is involved in the pathophysiology of autoimmune thyroiditis (AIT) remains to be elucidated. In the present study, the effects of the CXCR4 antagonist, AMD3100, in an iodine­induced autoimmune thyroiditis model were investigated. NOD.H­2h4 mice were randomly separated into a control, AIT and AIT+AMD3100 groups. The mice were fed with 0.05% sodium iodide water for 8 weeks to induce AIT. The AMD3100­treated mice were administered with the CXCR4 antagonist at a dose of 10 mg/kg intraperitoneally three times a week during the experimental period. The percentages of CD19+interleukin (IL)10+ B cells and CD4+IL10+ T cells, and the mRNA expression levels of IL10 in the splenocytes were reduced in the AIT group, compared with the control group, however, they increased following AMD3100 treatment, compared with the untreated AIT group. The percentages of CD4+ T cells, CD8+ T cells, CD19+ B cells and CD8+ interferon (IFN)γ+ T cells, and the mRNA expression levels of IFNγ increased in the AIT group, compared with the control group, however, these were reduced in the AMD3100 group, compared with the AIT group. The AMD3100­treated mice also had lower serum thyroglobulin antibody titers and reduced lymphocytic infiltration in the thyroid, compared with the untreated AIT mice. These results suggested that inhibition of this chemokine axis may offer potential as a therapeutic target for the treatment of AIT.

The thyroxine-containing thyroglobulin peptide (aa 2549-2560) is a target epitope in iodide-accelerated spontaneous autoimmune thyroiditis.

Enhanced iodide ingestion is known to accelerate the incidence and severity of spontaneous autoimmune thyroiditis [iodide-accelerated spontaneous autoimmune thyroiditis (ISAT)] in NOD.H2(h4) mice. CD4+ cells are required for the development and maintenance of ISAT, but their target epitopes remain unknown. In this study, we show that the previously identified thyroglobulin (Tg) T cell epitope p2549-2560 containing thyroxine at position 2553 (T4p2553) induces thyroiditis as well as strong specific T and B cell responses in NOD.H2(h4) mice. In ISAT, activated CD4+ T cells specific for T4p2553 are detected before the disease onset in thyroid-draining cervical lymph nodes only in mice placed on an iodide-rich diet and not in age-matched controls. In addition, selective enrichment of CD4+ IFN-γ+ T4p2553-specific cells is observed among cervical lymph node cells and intrathyroidal lymphocytes. T4p2553 was equally detectable on dendritic cells obtained ex vivo from cervical lymph node cells of NaI-fed or control mice, suggesting that the iodide-rich diet contributes to the activation of autoreactive cells rather than the generation of the autoantigenic epitope. Furthermore, spontaneous T4p2553-specific IgG are not detectable within the strong Tg-specific autoantibody response. To our knowledge, these data identify for the first time a Tg T cell epitope as a spontaneous target in ISAT.

Identification of nuclear spliceosomal antigens targeted by NOD mouse antibodies following sodium iodide intake.

The non-obese diabetic (NOD) mouse spontaneously develops a range of autoreactive responses including an autoantibody response to nuclear antigens. As elevated dietary iodine has been shown to increase thyroid autoimmune pathology in NOD mice, the effect of sodium iodide (NaI) on the development of anti-nuclear antibodies (ANA) was assessed. Interestingly, the NaI symporter is expressed in both thyroid and salivary glands. Elevated dietary iodine was found to increase the percentage of male NOD mice developing autoantibodies. Specifically, the nuclear autoantibodies that develop in NOD mice were shown to target specific spliceosomal components. The target specificity of the autoantibodies was determined using recombinant spliceosomal proteins and shown to include U1A, U170K, U2B'', U2A', as well as the Sm proteins D1, D2, and B. The autoantibody isotypes most consistently represented were IgG2a and IgG2b.

Both CD4(+) T cells and CD8(+) T cells are required for iodine accelerated thyroiditis in NOD mice.

The nonobese diabetic (NOD) mouse, a spontaneous animal model for insulin-dependent diabetes mellitus, displays a tendency in common with human diabetic populations to develop autoimmune thyroiditis although incidence and severity of thyroid lesions vary widely among different colonies around the world. A congenic strain of NOD mice bearing I-Ak on a NOD background (NOD-H2(h4)) has recently been derived and displays a much greater tendency to develop thyroiditis and autoantibodies to mouse thyroglobulin (MTg) although it is free of diabetes. Both thyroid infiltrates and autoantibody formation are accelerated and enhanced in NOD-H2(h4) mice by increased iodine intake. The effect of increased iodine intake on NOD mice themselves has not been directly investigated although a recent study of these animals given high or low doses of iodine showed no follicular destruction unless the mice were first rendered goitrous by iodine deprivation. We found that dietary iodine increased both the incidence and the severity of thyroid lesions in our NOD mice although autoantibodies to MTg were absent. NOD background genes appear to be essential for the development of these lesions, which were maximal after 4 weeks of iodine administration and showed no significant regression when the iodine was stopped. Furthermore, our studies show for the first time that both CD4(+) and CD8(+) T cells are necessary for the development of this accelerated but essentially spontaneous murine thyroid disease.

Comparison of serum cortisol concentrations in clinically normal dogs after administration of freshly reconstituted versus reconstituted and stored frozen cosyntropin.

OBJECTIVE: To evaluate the biological stability of reconstituted cosyntropin after storage at -20 C for 2, 4, and 6 months. DESIGN: Prospective study. ANIMALS: 10 clinically normal dogs. PROCEDURE: Serum cortisol concentrations in dogs were determined before and 1 hour after administration of freshly reconstituted cosyntropin (synthetic ACTH) or cosyntropin that had been reconstituted and stored frozen at -20 C in plastic syringes for 2, 4, and 6 months. Cosyntropin was administered at 5 micrograms/kg (2.3 micrograms/lb) of body weight, i.v. RESULTS: Baseline serum cortisol concentrations were similar at each sampling time. Compared with the effects of freshly reconstituted cosyntropin, administration of cosyntropin that had been frozen did not have a significantly different effect on serum cortisol concentrations. CLINICAL IMPLICATIONS: Cosyntropin can be reconstituted and stored frozen at -20 C in plastic syringes for 6 months with no adverse effects on bioactivity of the polypeptide.

Involvement of T cell immunity in the transient thyroid inflammation induced by iodide in goitrous BALB/C and nude mice.

To evaluate the involvement of T cell immunity in the thyroiditis induced in goitrous mice by iodide administration, we analyzed the immunological changes happening in the thyroid glands and lymph nodes during goiter involution in balb/c and athymic nude mice of similar background. In both balb/c and nude mice, goiter involution was characterized by thyroid cell necrosis and inflammation. In balb/c mice, the inflammatory infiltrate was made of numerous Ia+ cells. Their number was unchanged during goiter development, but was significantly increased after 2 days of involution and remained high after 8 days. CD4+ and CD8+ T cells were also observed, some of which were clearly activated since they expressed the receptor for Interleukin-2 (IL-2R). The numbers of CD4+, CD8+ and IL-2R+ T cells were increased during goiter as compared to control mice, and they reached a maximum at day 1 of involution. In nude mice, unexpectedly, CD4+ and CD8+ T cells were also found in the thyroid. Their numbers, as well as the number of Ia+ cells, were significantly increased at the onset of involution, but they remained systematically lower than in the corresponding groups of balb/c mice. Iodide treatment of goitrous mice also induced modifications of the lymph nodes draining the thyroid: enlargement of the paracortical T zone, presence of germinal centers in cortical follicles, and increase of the density of IL-2R+ cells. Mesenteric lymph nodes taken as controls were unchanged. Thus, three observations suggest the involvement of T cell immunity in iodine-induced thyroid inflammation: 1. Infiltration of Ia+, CD4+, CD8+ and IL-2R+ cells. 2. Signs of stimulation in thyroid lymph nodes, 3. Significant differences between balb/c and nude mice, in which the inflammatory reaction is weaker.

In vitro study of acute toxic effects of high iodide doses in human thyroid follicles.

The acute effects of increasing doses of sodium iodide were studied on human thyroid follicles isolated from normal paranodular tissue. After 24 h incubation in culture medium, follicles isolated from most thyroids maintained their capacity for 125I accumulation and organification and a normal cellular ultrastructure. 125I accumulation was significantly increased after addition of TSH, whereas 125I organification was not affected. In presence of TSH, numerous follicles had large empty-looking follicular lumina unlabeled on autoradiographies. Follicles incubated for 24 h in the presence of a low concentration (10(-7) M) of iodide retained their function and morphology. However, incubation with a high dose of iodide (10(-3) M) caused marked inhibition of 125I accumulation and organification reaching values similar to those obtained in presence of inhibitors of iodide trapping and organification. At high doses, iodide induced necrosis of thyroid epithelial cells: the percentage of necrotic cells was significantly increased with 10(-5) M and doubled with 10(-3) M as compared to values measured at 10(-7) M. Ultrastructural lesions such as apical blebbing, cytoplasmic fragments desquamation, endoplasmic reticulum vesiculation, and accumulation of lipofuscin in secondary lysosomes were also present. The necrotic effect and the ultrastructural alterations also occurred in the presence of TSH but were prevented by the addition of inhibitors of iodide trapping or organification. These results demonstrate a direct acute toxic effect of iodide in human thyroid cells. The nature of the ultrastructural alterations is in agreement with a mechanism of toxicity involving a free radical attack and lipid peroxidation as observed in other tissues.

[Comparative biological effectiveness of 125I and 131I]. / Sravnitel'naia biologicheskaia éffektivnost' 125I i 131I.

An experimental study of comparative biological efficacy of 125I and 131I at equal absorbed doses (2.3 and 13 Gy) in the thyroid of rats showed that the blastomogenic effect of both radionuclides was almost the same. Sex differences were found in the development of long-term consequences (20 mos. after drug administration): breast tumors developed in female rats and thyroid tumors in male rats.