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Germline deletion of β2 microglobulin or CD1d reduces anti-phospholipid antibody, but increases autoantibodies against non-phospholipid antigens in the NZB/W F1 model of lupus.

Singh RR, Yang JQ, Kim PJ, Halder RC - Arthritis Res. Ther. (2013)

Bottom Line: Since CD1d is known to bind and present phospholipid antigens to T cells, we asked if the deficiency of β2m or CD1d will impact the development of anti-phospholipid antibodies as compared to other aspects of lupus autoimmunity.Whereas β2m° BWF1 mice had reduced serum IgG, they had increased mortality, nephritis, serum IgG anti-DNA antibody and RF as compared to heterozygous and wild-type littermates.Such CD1d dependence of anti-CL antibody production is not mediated by CD1d/glycolipid-reactive iNKT cells, as these cells reduced the production of RF and anti-DNA antibodies but had no effect on anti-CL antibodies.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Introduction: β2-microglobulin (β2m) is required for the surface expression of MHC class I and class I-like proteins such as CD1d, Qa1 and neonatal Fc receptor (FcRn), all of which may impact the development of autoimmunity. Since CD1d is known to bind and present phospholipid antigens to T cells, we asked if the deficiency of β2m or CD1d will impact the development of anti-phospholipid antibodies as compared to other aspects of lupus autoimmunity.

Methods: We introgressed the β2m- genotype onto the NZB and NZW backgrounds for 12 to 14 generations to generate genetically lupus-susceptible (NZB/NZW)F1 (BWF1) mice that are β2m-deficient (β2m°). Circulating immunoglobulins (Ig), rheumatoid factor (RF), anti-DNA and anti-cardiolipin (anti-CL) antibodies, and renal disease were analyzed in these and CD1d-deficient (CD1d°) BWF1 mice that we had previously generated.

Results: Whereas β2m° BWF1 mice had reduced serum IgG, they had increased mortality, nephritis, serum IgG anti-DNA antibody and RF as compared to heterozygous and wild-type littermates. These effects were recapitulated in CD1d° BWF1 mice, except that they also had increased serum IgG as compared to control littermates. Intriguingly, both β2m° and CD1d° mice had lower serum anti-CL antibody levels than in control littermates. Such CD1d dependence of anti-CL antibody production is not mediated by CD1d/glycolipid-reactive iNKT cells, as these cells reduced the production of RF and anti-DNA antibodies but had no effect on anti-CL antibodies.

Conclusions: We report a novel dichotomous role of β2m and CD1d, whereby these molecules differently regulate autoimmunity against phospholipid versus non-phospholipid autoantigens.

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Activated invariant natural killer T (iNKT) cells reduce anti-DNA antibodies, but not anti-cardiolipin (anti-CL) antibodies. (a) In vitro studies. Spleen cells harvested from 5-month-old BWF1 mice were cultured with αGalCer for 5 days, and supernatants tested for immunoglobulin G (IgG) anti-DNA and anti-CL antibodies, shown as the mean ± SD optical density (OD); *P <0.05 to 0.01, n = 5. Results represent two similar experiments. (b) In vivo studies. BALB/c SCID mice (4-month-old) were reconstituted with purified B-cells from iNKT cell-deficient Jα18° BALB/c mice, and injected with lipopolysaccharide and αGalCer, as described in Methods. These mice were then implanted with T-cells from Vα14Tg BALB/c mice or with T-cells from Jα18° BALB/c mice. Four days after the reconstitution, spleen cells were harvested and cultured without any further stimulation for 6 days. Culture supernatants were tested for IgG anti-DNA and anti-CL antibodies. Results represent two independent experiments, each using three mice per group. *P <0.05.
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Figure 6: Activated invariant natural killer T (iNKT) cells reduce anti-DNA antibodies, but not anti-cardiolipin (anti-CL) antibodies. (a) In vitro studies. Spleen cells harvested from 5-month-old BWF1 mice were cultured with αGalCer for 5 days, and supernatants tested for immunoglobulin G (IgG) anti-DNA and anti-CL antibodies, shown as the mean ± SD optical density (OD); *P <0.05 to 0.01, n = 5. Results represent two similar experiments. (b) In vivo studies. BALB/c SCID mice (4-month-old) were reconstituted with purified B-cells from iNKT cell-deficient Jα18° BALB/c mice, and injected with lipopolysaccharide and αGalCer, as described in Methods. These mice were then implanted with T-cells from Vα14Tg BALB/c mice or with T-cells from Jα18° BALB/c mice. Four days after the reconstitution, spleen cells were harvested and cultured without any further stimulation for 6 days. Culture supernatants were tested for IgG anti-DNA and anti-CL antibodies. Results represent two independent experiments, each using three mice per group. *P <0.05.

Mentions: CD1d-restricted T cells comprise glycolipid-reactive iNKT cells that express the invariant TCR Vα14Jα18 and other NKT cells that do not express the invariant TCR. To determine the effect of iNKT cells on various autoantibodies, we cultured BWF1 spleen cells with glycolipid αGalCer. We found that while IgG anti-DNA antibody levels were reduced in the presence of αGalCer, IgG anti-CL antibody levels were unaffected (Figure 6a).


Germline deletion of β2 microglobulin or CD1d reduces anti-phospholipid antibody, but increases autoantibodies against non-phospholipid antigens in the NZB/W F1 model of lupus.

Singh RR, Yang JQ, Kim PJ, Halder RC - Arthritis Res. Ther. (2013)

Activated invariant natural killer T (iNKT) cells reduce anti-DNA antibodies, but not anti-cardiolipin (anti-CL) antibodies. (a) In vitro studies. Spleen cells harvested from 5-month-old BWF1 mice were cultured with αGalCer for 5 days, and supernatants tested for immunoglobulin G (IgG) anti-DNA and anti-CL antibodies, shown as the mean ± SD optical density (OD); *P <0.05 to 0.01, n = 5. Results represent two similar experiments. (b) In vivo studies. BALB/c SCID mice (4-month-old) were reconstituted with purified B-cells from iNKT cell-deficient Jα18° BALB/c mice, and injected with lipopolysaccharide and αGalCer, as described in Methods. These mice were then implanted with T-cells from Vα14Tg BALB/c mice or with T-cells from Jα18° BALB/c mice. Four days after the reconstitution, spleen cells were harvested and cultured without any further stimulation for 6 days. Culture supernatants were tested for IgG anti-DNA and anti-CL antibodies. Results represent two independent experiments, each using three mice per group. *P <0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3672782&req=5

Figure 6: Activated invariant natural killer T (iNKT) cells reduce anti-DNA antibodies, but not anti-cardiolipin (anti-CL) antibodies. (a) In vitro studies. Spleen cells harvested from 5-month-old BWF1 mice were cultured with αGalCer for 5 days, and supernatants tested for immunoglobulin G (IgG) anti-DNA and anti-CL antibodies, shown as the mean ± SD optical density (OD); *P <0.05 to 0.01, n = 5. Results represent two similar experiments. (b) In vivo studies. BALB/c SCID mice (4-month-old) were reconstituted with purified B-cells from iNKT cell-deficient Jα18° BALB/c mice, and injected with lipopolysaccharide and αGalCer, as described in Methods. These mice were then implanted with T-cells from Vα14Tg BALB/c mice or with T-cells from Jα18° BALB/c mice. Four days after the reconstitution, spleen cells were harvested and cultured without any further stimulation for 6 days. Culture supernatants were tested for IgG anti-DNA and anti-CL antibodies. Results represent two independent experiments, each using three mice per group. *P <0.05.
Mentions: CD1d-restricted T cells comprise glycolipid-reactive iNKT cells that express the invariant TCR Vα14Jα18 and other NKT cells that do not express the invariant TCR. To determine the effect of iNKT cells on various autoantibodies, we cultured BWF1 spleen cells with glycolipid αGalCer. We found that while IgG anti-DNA antibody levels were reduced in the presence of αGalCer, IgG anti-CL antibody levels were unaffected (Figure 6a).

Bottom Line: Since CD1d is known to bind and present phospholipid antigens to T cells, we asked if the deficiency of β2m or CD1d will impact the development of anti-phospholipid antibodies as compared to other aspects of lupus autoimmunity.Whereas β2m° BWF1 mice had reduced serum IgG, they had increased mortality, nephritis, serum IgG anti-DNA antibody and RF as compared to heterozygous and wild-type littermates.Such CD1d dependence of anti-CL antibody production is not mediated by CD1d/glycolipid-reactive iNKT cells, as these cells reduced the production of RF and anti-DNA antibodies but had no effect on anti-CL antibodies.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Introduction: β2-microglobulin (β2m) is required for the surface expression of MHC class I and class I-like proteins such as CD1d, Qa1 and neonatal Fc receptor (FcRn), all of which may impact the development of autoimmunity. Since CD1d is known to bind and present phospholipid antigens to T cells, we asked if the deficiency of β2m or CD1d will impact the development of anti-phospholipid antibodies as compared to other aspects of lupus autoimmunity.

Methods: We introgressed the β2m- genotype onto the NZB and NZW backgrounds for 12 to 14 generations to generate genetically lupus-susceptible (NZB/NZW)F1 (BWF1) mice that are β2m-deficient (β2m°). Circulating immunoglobulins (Ig), rheumatoid factor (RF), anti-DNA and anti-cardiolipin (anti-CL) antibodies, and renal disease were analyzed in these and CD1d-deficient (CD1d°) BWF1 mice that we had previously generated.

Results: Whereas β2m° BWF1 mice had reduced serum IgG, they had increased mortality, nephritis, serum IgG anti-DNA antibody and RF as compared to heterozygous and wild-type littermates. These effects were recapitulated in CD1d° BWF1 mice, except that they also had increased serum IgG as compared to control littermates. Intriguingly, both β2m° and CD1d° mice had lower serum anti-CL antibody levels than in control littermates. Such CD1d dependence of anti-CL antibody production is not mediated by CD1d/glycolipid-reactive iNKT cells, as these cells reduced the production of RF and anti-DNA antibodies but had no effect on anti-CL antibodies.

Conclusions: We report a novel dichotomous role of β2m and CD1d, whereby these molecules differently regulate autoimmunity against phospholipid versus non-phospholipid autoantigens.

Show MeSH
Related in: MedlinePlus