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. 2003 Apr;62(4):341–346. doi: 10.1136/ard.62.4.341

Beneficial effects of the anti-oestrogen tamoxifen on systemic lupus erythematosus of (NZBxNZW)F1 female mice are associated with specific reduction of IgG3 autoantibodies

Z Sthoeger 1, H Zinger 1, E Mozes 1
PMCID: PMC1754513  PMID: 12634234

Abstract

Background: Sex hormones have been shown to influence the immune system and to modify the course of autoimmune disorders.

Objective: To examine the effects of the oestrogen antagonist tamoxifen on the course of systemic lupus erythematosus (SLE) in (NZBxNZW)F1 mice.

Methods: Groups of 8 week old (NZBxNZW)F1 female mice were treated with tamoxifen (800 µg/mouse; twice a week) or with double distilled water for four months. Mice were evaluated monthly for the presence of autoantibodies directed against DNA and nuclear extract (NE) by enzyme linked immunosorbent assay (ELISA). White blood cells and thrombocytes were quantified by a cell counter and proteinuria by combistix kit. At 6 months of age, all mice that did not die spontaneously were killed and evaluated for the presence of glomerular immune deposits by indirect immunofluorescence assay. IgG isotypes of autoantibodies in the mouse sera and glomeruli were determined by γ chain specific antibodies.

Results: Tamoxifen treatment significantly reduced autoantibody production directed against either NE or DNA. The latter reduction was mainly in autoantibodies of the IgG3 isotype. Furthermore, tamoxifen had significant beneficial effects on the course of SLE in (NZBxNZW)F1 mice. At 6 months of age, 40% of the untreated mice died spontaneously, whereas all the tamoxifen treated mice were still alive. All untreated mice showed severe thrombocytopenia and persistent proteinuria, with diffuse glomerular immune deposits of IgG2a and IgG3 isotypes in their kidneys. In contrast, the tamoxifen treated mice had a normal number of thrombocytes and only minimal proteinuria. Moreover, glomerular immune deposits were detected in <40% of the tamoxifen treated mice. The latter were mainly of the IgG2a but not of the IgG3 isotype.

Conclusion: The results clearly show the remarkable therapeutic effects of tamoxifen on SLE of (NZBxNZW)F1 female mice and suggest that these beneficial effects are related to the specific reduction of IgG3 autoantibodies.

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Figure 1 .

Figure 1

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Autoantibodies of 6 month old (NZBxNZW)F1 female mice. Tamoxifen treated and untreated mice were bled, at 6 months of age, and the reactivity of their sera (total IgG) with dsDNA (A) and NE (B) was determined. Results represent mean (SEM) OD values (at 414 nm) of each individual mouse obtained from one representative of three consecutive experiments. Note the differences in sera dilution between the two assays.

Figure 2 .

Figure 2

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IgG isotypes of anti-dsDNA autoantibodies of 6 month old (NZBxNZW)F1 female mice. Tamoxifen treated and untreated mice were bled at 6 months of age and the isotypes of the anti-dsDNA autoantibodies in the mouse sera (mean (SEM) of each individual mouse) were determined by using γ1, γ2a, γ2b, and γ3 chain specific goat antimouse antibodies. Results of one representative out of three consecutive experiments are presented.

Figure 3 .

Figure 3

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Immunohistology of kidney sections obtained from (NZBxNZW)F1 female mice. Kidney sections were obtained from 2 month old control mice (1A, 1B, 1C), 6 month old untreated mice (2A, 2B, 2C) and 6 month old tamoxifen treated (3A, 3B, 3C, 4A, 4B, 4C) (NZBxNZW)F1 mice. Mice were killed and their kidney sections (5 µm) were stained with FITC labelled goat antimouse IgG (total IgG; panel A), IgG2a (panel B), or IgG3 (panel C) using γ, γ2a, or γ3 heavy chain specific antibodies, respectively. Representative figures (x200) are presented. Sections 1 and 4 were defined as negative (score 0), section 2 as strongly positive (+++) and section 3 as positive (+/++). Note the relatively low intensity of IgG3 deposits in kidney sections obtained from tamoxifen treated mice (3C).

Selected References

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