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Antinuclear autoantibodies and lupus nephritis in transgenic mice expressing interferon gamma in the epidermis.

Seery JP, Carroll JM, Cattell V, Watt FM - J. Exp. Med. (1997)

Bottom Line: The mechanism and site of production of these autoantibodies is unknown, but there is evidence that interferon (IFN) gamma plays a key role.There was no evidence of organ-specific autoimmunity, but transgenic animals produced autoantibodies against dsDNA and histones.Our findings are consistent with a central role for the skin immune system, acting under the influence of IFN-gamma, in the pathogenesis of SLE.

View Article: PubMed Central - PubMed

Affiliation: Keratinocyte Laboratory, Imperial Cancer Research Fund, London WC2A 3PX, United Kingdom.

ABSTRACT
Systemic lupus erythematosus (SLE) is a potentially fatal non-organ-specific autoimmune disease that predominantly affects women. Features of the disease include inflammatory skin lesions and widespread organ damage caused by deposition of anti-dsDNA autoantibodies. The mechanism and site of production of these autoantibodies is unknown, but there is evidence that interferon (IFN) gamma plays a key role. We have used the involucrin promoter to overexpress IFN-gamma in the suprabasal layers of transgenic mouse epidermis. There was no evidence of organ-specific autoimmunity, but transgenic animals produced autoantibodies against dsDNA and histones. Autoantibody levels in female mice were significantly higher than in male transgenic mice. Furthermore, there was IgG deposition in the glomeruli of all female mice and histological evidence of severe proliferative glomerulonephritis in a proportion of these animals. Our findings are consistent with a central role for the skin immune system, acting under the influence of IFN-gamma, in the pathogenesis of SLE.

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Screening serum for anti-dsDNA and antihistone autoantibodies. (A) C. luciliae assay. Staining with serum from a transgenic mouse results in  fluorescence of the kinetoplast (arrow) of each organism. A series of 1-μm optical sections through the specimens was obtained with a confocal microscope and a composite image (Z series) was constructed. Scale bar: 10 μm. (B) Anti-dsDNA and (C) antihistone autoantibody levels in transgenic mouse  serum. Sera from 17 transgenic mice and 6 negative littermate controls were tested individually against dsDNA at 1:250 and histones at 1:1,000 dilution.  The OD value for each sample represents the mean of three measurements. LC, Littermate controls; TM, transgenic males; TF, transgenic females. Four  MRL/lpr mice known to produce high levels of antinuclear antibodies are included as positive controls. A serum sample known to be positive for anti-ssDNA showed no significant binding in the dsDNA ELISA (OD 0.00 ± 0.05). All sera were tested on uncoated ELISA plate plastic. Nonspecific binding to plastic was low (OD on uncoated wells ranged from 0 to 5% of values on antigen-coated wells).
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Figure 2: Screening serum for anti-dsDNA and antihistone autoantibodies. (A) C. luciliae assay. Staining with serum from a transgenic mouse results in fluorescence of the kinetoplast (arrow) of each organism. A series of 1-μm optical sections through the specimens was obtained with a confocal microscope and a composite image (Z series) was constructed. Scale bar: 10 μm. (B) Anti-dsDNA and (C) antihistone autoantibody levels in transgenic mouse serum. Sera from 17 transgenic mice and 6 negative littermate controls were tested individually against dsDNA at 1:250 and histones at 1:1,000 dilution. The OD value for each sample represents the mean of three measurements. LC, Littermate controls; TM, transgenic males; TF, transgenic females. Four MRL/lpr mice known to produce high levels of antinuclear antibodies are included as positive controls. A serum sample known to be positive for anti-ssDNA showed no significant binding in the dsDNA ELISA (OD 0.00 ± 0.05). All sera were tested on uncoated ELISA plate plastic. Nonspecific binding to plastic was low (OD on uncoated wells ranged from 0 to 5% of values on antigen-coated wells).

Mentions: To examine whether antibodies to dsDNA were present, we screened serum for reactivity to the kineoplast of the flagellate organism C. luciliae. Indirect immunofluorescence testing of serum on C. luciliae has been used as a specific test for the presence of anti-dsDNA autoantibodies (22). Serum samples from 21 transgenic mice and 12 negative littermate controls were tested. 18 samples from transgenic animals produced definite staining of the kinetoplast (see Fig. 2 A and Table 1). All littermate controls were negative on this test (Table 2).


Antinuclear autoantibodies and lupus nephritis in transgenic mice expressing interferon gamma in the epidermis.

Seery JP, Carroll JM, Cattell V, Watt FM - J. Exp. Med. (1997)

Screening serum for anti-dsDNA and antihistone autoantibodies. (A) C. luciliae assay. Staining with serum from a transgenic mouse results in  fluorescence of the kinetoplast (arrow) of each organism. A series of 1-μm optical sections through the specimens was obtained with a confocal microscope and a composite image (Z series) was constructed. Scale bar: 10 μm. (B) Anti-dsDNA and (C) antihistone autoantibody levels in transgenic mouse  serum. Sera from 17 transgenic mice and 6 negative littermate controls were tested individually against dsDNA at 1:250 and histones at 1:1,000 dilution.  The OD value for each sample represents the mean of three measurements. LC, Littermate controls; TM, transgenic males; TF, transgenic females. Four  MRL/lpr mice known to produce high levels of antinuclear antibodies are included as positive controls. A serum sample known to be positive for anti-ssDNA showed no significant binding in the dsDNA ELISA (OD 0.00 ± 0.05). All sera were tested on uncoated ELISA plate plastic. Nonspecific binding to plastic was low (OD on uncoated wells ranged from 0 to 5% of values on antigen-coated wells).
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Related In: Results  -  Collection

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Figure 2: Screening serum for anti-dsDNA and antihistone autoantibodies. (A) C. luciliae assay. Staining with serum from a transgenic mouse results in fluorescence of the kinetoplast (arrow) of each organism. A series of 1-μm optical sections through the specimens was obtained with a confocal microscope and a composite image (Z series) was constructed. Scale bar: 10 μm. (B) Anti-dsDNA and (C) antihistone autoantibody levels in transgenic mouse serum. Sera from 17 transgenic mice and 6 negative littermate controls were tested individually against dsDNA at 1:250 and histones at 1:1,000 dilution. The OD value for each sample represents the mean of three measurements. LC, Littermate controls; TM, transgenic males; TF, transgenic females. Four MRL/lpr mice known to produce high levels of antinuclear antibodies are included as positive controls. A serum sample known to be positive for anti-ssDNA showed no significant binding in the dsDNA ELISA (OD 0.00 ± 0.05). All sera were tested on uncoated ELISA plate plastic. Nonspecific binding to plastic was low (OD on uncoated wells ranged from 0 to 5% of values on antigen-coated wells).
Mentions: To examine whether antibodies to dsDNA were present, we screened serum for reactivity to the kineoplast of the flagellate organism C. luciliae. Indirect immunofluorescence testing of serum on C. luciliae has been used as a specific test for the presence of anti-dsDNA autoantibodies (22). Serum samples from 21 transgenic mice and 12 negative littermate controls were tested. 18 samples from transgenic animals produced definite staining of the kinetoplast (see Fig. 2 A and Table 1). All littermate controls were negative on this test (Table 2).

Bottom Line: The mechanism and site of production of these autoantibodies is unknown, but there is evidence that interferon (IFN) gamma plays a key role.There was no evidence of organ-specific autoimmunity, but transgenic animals produced autoantibodies against dsDNA and histones.Our findings are consistent with a central role for the skin immune system, acting under the influence of IFN-gamma, in the pathogenesis of SLE.

View Article: PubMed Central - PubMed

Affiliation: Keratinocyte Laboratory, Imperial Cancer Research Fund, London WC2A 3PX, United Kingdom.

ABSTRACT
Systemic lupus erythematosus (SLE) is a potentially fatal non-organ-specific autoimmune disease that predominantly affects women. Features of the disease include inflammatory skin lesions and widespread organ damage caused by deposition of anti-dsDNA autoantibodies. The mechanism and site of production of these autoantibodies is unknown, but there is evidence that interferon (IFN) gamma plays a key role. We have used the involucrin promoter to overexpress IFN-gamma in the suprabasal layers of transgenic mouse epidermis. There was no evidence of organ-specific autoimmunity, but transgenic animals produced autoantibodies against dsDNA and histones. Autoantibody levels in female mice were significantly higher than in male transgenic mice. Furthermore, there was IgG deposition in the glomeruli of all female mice and histological evidence of severe proliferative glomerulonephritis in a proportion of these animals. Our findings are consistent with a central role for the skin immune system, acting under the influence of IFN-gamma, in the pathogenesis of SLE.

Show MeSH
Related in: MedlinePlus