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Murine lupus susceptibility locus Sle1a requires the expression of two sub-loci to induce inflammatory T cells.

Cuda CM, Zeumer L, Sobel ES, Croker BP, Morel L - Genes Immun. (2010)

Bottom Line: Sle1a.1, and to a lesser extent Sle1a.2, significantly affected CD4(+) T-cell activation as well as Treg differentiation and function.As the Sle1a.1 and Sle1a.2 intervals contain only 1 and 15 known genes, respectively, this study considerably reduces the number of candidate genes responsible for the production of autoreactive T cells.These results also show that the Sle1 locus is an excellent model for the genetic architecture of lupus, in which a major obligate phenotype results from the coexpression of multiple genetic variants with individual weak effects.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Immunology, and Laboratory Medicine, University of Florida, Gainesville, 32610-0275, USA.

ABSTRACT
The NZM2410-derived Sle1a lupus susceptibility locus induces activated autoreactive CD4(+) T cells and reduces the number and function of Foxp3(+) regulatory T cells (Tregs). In this study, we first showed that Sle1a contributes to autoimmunity by increasing antinuclear antibody production when expressed on either NZB or NZW heterozygous genomes, and by enhancing the chronic graft versus host disease response indicating an expansion of the autoreactive B-cell pool. Screening two non-overlapping recombinants, the Sle1a.1 and Sle1a.2 intervals that cover the entire Sle1a locus, revealed that both Sle1a.1 and Sle1a.2 were necessary for the full Sle1a phenotype. Sle1a.1, and to a lesser extent Sle1a.2, significantly affected CD4(+) T-cell activation as well as Treg differentiation and function. Sle1a.2 also increased the production of autoreactive B cells. As the Sle1a.1 and Sle1a.2 intervals contain only 1 and 15 known genes, respectively, this study considerably reduces the number of candidate genes responsible for the production of autoreactive T cells. These results also show that the Sle1 locus is an excellent model for the genetic architecture of lupus, in which a major obligate phenotype results from the coexpression of multiple genetic variants with individual weak effects.

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Sle1a Tregs and Teffs induce colon and renal inflammatory infiltrates. A. Colon from a B6.Rag mouse reconstituted with B6 Teff: B6 Treg, which is indistinguishable from unmanipulated B6. Lumen is toward the right of figure and the arrow head marks muscularis mucosa. B. High grade colitis in a mouse reconstituted with Sle1a Teff alone. Arrow head points to muscularis mucosa as in A. The wall is markedly expanded with lymphocytic cryptitis (thin arrow), inflammation of the lamina propria with occasional giant cells (upper right, asterisk), inflammation of the muscularis mucosa, and interstitial submucosa on left of picture. C. Kidney cortex from a B6.Rag mouse reconstituted with B6 Teff: B6 Treg, which is indistinguishable from unmanipulated B6. D. Kidney from a mouse reconstituted with Sle1a Teff alone, with a moderate size focus of interstitial inflammation (center between glomerulus and vein in upper center). Epithelioid giant cell (asterisk) is shown in set (1000x magnification). E. Normal tubules a mouse reconstituted with B6 Teff: B6 Treg, with open round nuclei in epithelial cells and peritubular capillary (arrowhead). F. Tubules from same animal as in D, showing tubulitis with lymphocytic infiltration containing dark, dense oval nuclei (arrows and others) and capillaritis in peritubular capillaries congested with lymphocytes (between tubules with arrows). Dilated but otherwise empty capillaries are also present (e.g. arrowhead). A–D. 400x original magnification, E–F. 1000x original magnification. A–B. H&E stain; C–F. PAS stain. G. Histological assessment of colitis ranked among the 6 groups with Teffs:Tregs of mixed origin. **: p<0.01 indicate the significance of comparisons with the B6:B6 group in a Dunnett’s Multiple Comparison Test. ##: p < 0.0004, t test between Sle1a1 Tregs:B6 Teffs and B6 Tregs:Sle1a.1 Teffs. H. Number of renal foci induced by Teffs as compared to controls B6:B6. The values of Teffs alone are significantly higher than that of B6:B6 for all strains. **: p<0.01 indicate the significance of the comparison between B6 and Sle1a Teffs. I. Number of renal foci induced Teffs:Tregs of mixed origin *: p<0.05, **: p<0.01 indicate the significance of comparisons with the B6:B6 group in a Dunnett’s Multiple Comparison Test.
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Figure 6: Sle1a Tregs and Teffs induce colon and renal inflammatory infiltrates. A. Colon from a B6.Rag mouse reconstituted with B6 Teff: B6 Treg, which is indistinguishable from unmanipulated B6. Lumen is toward the right of figure and the arrow head marks muscularis mucosa. B. High grade colitis in a mouse reconstituted with Sle1a Teff alone. Arrow head points to muscularis mucosa as in A. The wall is markedly expanded with lymphocytic cryptitis (thin arrow), inflammation of the lamina propria with occasional giant cells (upper right, asterisk), inflammation of the muscularis mucosa, and interstitial submucosa on left of picture. C. Kidney cortex from a B6.Rag mouse reconstituted with B6 Teff: B6 Treg, which is indistinguishable from unmanipulated B6. D. Kidney from a mouse reconstituted with Sle1a Teff alone, with a moderate size focus of interstitial inflammation (center between glomerulus and vein in upper center). Epithelioid giant cell (asterisk) is shown in set (1000x magnification). E. Normal tubules a mouse reconstituted with B6 Teff: B6 Treg, with open round nuclei in epithelial cells and peritubular capillary (arrowhead). F. Tubules from same animal as in D, showing tubulitis with lymphocytic infiltration containing dark, dense oval nuclei (arrows and others) and capillaritis in peritubular capillaries congested with lymphocytes (between tubules with arrows). Dilated but otherwise empty capillaries are also present (e.g. arrowhead). A–D. 400x original magnification, E–F. 1000x original magnification. A–B. H&E stain; C–F. PAS stain. G. Histological assessment of colitis ranked among the 6 groups with Teffs:Tregs of mixed origin. **: p<0.01 indicate the significance of comparisons with the B6:B6 group in a Dunnett’s Multiple Comparison Test. ##: p < 0.0004, t test between Sle1a1 Tregs:B6 Teffs and B6 Tregs:Sle1a.1 Teffs. H. Number of renal foci induced by Teffs as compared to controls B6:B6. The values of Teffs alone are significantly higher than that of B6:B6 for all strains. **: p<0.01 indicate the significance of the comparison between B6 and Sle1a Teffs. I. Number of renal foci induced Teffs:Tregs of mixed origin *: p<0.05, **: p<0.01 indicate the significance of comparisons with the B6:B6 group in a Dunnett’s Multiple Comparison Test.

Mentions: Using the experimental colitis model,27 we have shown that Sle1a Tregs did not suppress tissue inflammation induced by either B6 or B6.Sle1a Teffs, and that Sle1a Teffs were resistant to suppression.20 We repeated this experiment to assess the in vivo effect of Sle1a.1 or Sle1a.2 expression on Teff and Treg functions, with B6.Ragmice receiving CD4+ CD25− Teffs from B6 or congenic (B6.Sle1a, B6.Sle1a.1 or B6.Sle1a.2) mice in the presence or absence of CD4+ CD25+ Tregs from these mice. As expected, the B6 Teff-induced colitis, defined by a 15% body weight loss and by histology, was completely abrogated by B6 Tregs (Fig. 6A), and the most severe colitis was induced by Sle1a Teff alone (Fig. 6B). A spectrum of severity was observed in mice receiving Sle1a.1 or Sle1a.2 Teffs and Tregs in combination with B6 T cells. The lesser involved colons showed inflammation limited to the crypts (cryptitis) and lamina propria. Increasing inflammation involved the muscularis mucosa and inner submucosal connective tissue stroma. The most severe inflammation involved the muscularis propria, submucosal stroma and serosa. The capillaries and venules in each layer with inflammation showed margination and congestion with mononuclear cells and swelling. Cryptitis was primarily lymphocytic but higher-grade specimens also had focal neutrophilic crypt abscesses. More epithelioid inflammation was also present in the higher-grade lesions in the lamina propria and deeper layers with nodules and occasional multinucleated epithelioid giant cells (Fig. 6B). Increasing inflammation was associated with increasing wall thickness up to three-fold normal (data not shown). Sle1a or Sle1a.1 Tregs did not suppress B6 Teff-induced colitis as well as B6 Tregs (Fig. 6G). Impaired Sle1a.2 Tregs were detected clinically with body weight loss (data not shown), but not by histology (Fig. 6G), indicating that this functional impairment was not as severe as for Sle1a.1. Sle1a.1 Teffs were resistant to suppression by B6 Tregs to a similar level as Sle1a Teffs (Fig. 6G). In contrast, mice reconstituted with B6 Tregs:Sle1a.2 Teffs or B6 Tregs:B6 Teffs had equivalent scores, indicating that Sle1a.2 does not increase Teff inflammatory functions in vivo.


Murine lupus susceptibility locus Sle1a requires the expression of two sub-loci to induce inflammatory T cells.

Cuda CM, Zeumer L, Sobel ES, Croker BP, Morel L - Genes Immun. (2010)

Sle1a Tregs and Teffs induce colon and renal inflammatory infiltrates. A. Colon from a B6.Rag mouse reconstituted with B6 Teff: B6 Treg, which is indistinguishable from unmanipulated B6. Lumen is toward the right of figure and the arrow head marks muscularis mucosa. B. High grade colitis in a mouse reconstituted with Sle1a Teff alone. Arrow head points to muscularis mucosa as in A. The wall is markedly expanded with lymphocytic cryptitis (thin arrow), inflammation of the lamina propria with occasional giant cells (upper right, asterisk), inflammation of the muscularis mucosa, and interstitial submucosa on left of picture. C. Kidney cortex from a B6.Rag mouse reconstituted with B6 Teff: B6 Treg, which is indistinguishable from unmanipulated B6. D. Kidney from a mouse reconstituted with Sle1a Teff alone, with a moderate size focus of interstitial inflammation (center between glomerulus and vein in upper center). Epithelioid giant cell (asterisk) is shown in set (1000x magnification). E. Normal tubules a mouse reconstituted with B6 Teff: B6 Treg, with open round nuclei in epithelial cells and peritubular capillary (arrowhead). F. Tubules from same animal as in D, showing tubulitis with lymphocytic infiltration containing dark, dense oval nuclei (arrows and others) and capillaritis in peritubular capillaries congested with lymphocytes (between tubules with arrows). Dilated but otherwise empty capillaries are also present (e.g. arrowhead). A–D. 400x original magnification, E–F. 1000x original magnification. A–B. H&E stain; C–F. PAS stain. G. Histological assessment of colitis ranked among the 6 groups with Teffs:Tregs of mixed origin. **: p<0.01 indicate the significance of comparisons with the B6:B6 group in a Dunnett’s Multiple Comparison Test. ##: p < 0.0004, t test between Sle1a1 Tregs:B6 Teffs and B6 Tregs:Sle1a.1 Teffs. H. Number of renal foci induced by Teffs as compared to controls B6:B6. The values of Teffs alone are significantly higher than that of B6:B6 for all strains. **: p<0.01 indicate the significance of the comparison between B6 and Sle1a Teffs. I. Number of renal foci induced Teffs:Tregs of mixed origin *: p<0.05, **: p<0.01 indicate the significance of comparisons with the B6:B6 group in a Dunnett’s Multiple Comparison Test.
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Related In: Results  -  Collection

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Figure 6: Sle1a Tregs and Teffs induce colon and renal inflammatory infiltrates. A. Colon from a B6.Rag mouse reconstituted with B6 Teff: B6 Treg, which is indistinguishable from unmanipulated B6. Lumen is toward the right of figure and the arrow head marks muscularis mucosa. B. High grade colitis in a mouse reconstituted with Sle1a Teff alone. Arrow head points to muscularis mucosa as in A. The wall is markedly expanded with lymphocytic cryptitis (thin arrow), inflammation of the lamina propria with occasional giant cells (upper right, asterisk), inflammation of the muscularis mucosa, and interstitial submucosa on left of picture. C. Kidney cortex from a B6.Rag mouse reconstituted with B6 Teff: B6 Treg, which is indistinguishable from unmanipulated B6. D. Kidney from a mouse reconstituted with Sle1a Teff alone, with a moderate size focus of interstitial inflammation (center between glomerulus and vein in upper center). Epithelioid giant cell (asterisk) is shown in set (1000x magnification). E. Normal tubules a mouse reconstituted with B6 Teff: B6 Treg, with open round nuclei in epithelial cells and peritubular capillary (arrowhead). F. Tubules from same animal as in D, showing tubulitis with lymphocytic infiltration containing dark, dense oval nuclei (arrows and others) and capillaritis in peritubular capillaries congested with lymphocytes (between tubules with arrows). Dilated but otherwise empty capillaries are also present (e.g. arrowhead). A–D. 400x original magnification, E–F. 1000x original magnification. A–B. H&E stain; C–F. PAS stain. G. Histological assessment of colitis ranked among the 6 groups with Teffs:Tregs of mixed origin. **: p<0.01 indicate the significance of comparisons with the B6:B6 group in a Dunnett’s Multiple Comparison Test. ##: p < 0.0004, t test between Sle1a1 Tregs:B6 Teffs and B6 Tregs:Sle1a.1 Teffs. H. Number of renal foci induced by Teffs as compared to controls B6:B6. The values of Teffs alone are significantly higher than that of B6:B6 for all strains. **: p<0.01 indicate the significance of the comparison between B6 and Sle1a Teffs. I. Number of renal foci induced Teffs:Tregs of mixed origin *: p<0.05, **: p<0.01 indicate the significance of comparisons with the B6:B6 group in a Dunnett’s Multiple Comparison Test.
Mentions: Using the experimental colitis model,27 we have shown that Sle1a Tregs did not suppress tissue inflammation induced by either B6 or B6.Sle1a Teffs, and that Sle1a Teffs were resistant to suppression.20 We repeated this experiment to assess the in vivo effect of Sle1a.1 or Sle1a.2 expression on Teff and Treg functions, with B6.Ragmice receiving CD4+ CD25− Teffs from B6 or congenic (B6.Sle1a, B6.Sle1a.1 or B6.Sle1a.2) mice in the presence or absence of CD4+ CD25+ Tregs from these mice. As expected, the B6 Teff-induced colitis, defined by a 15% body weight loss and by histology, was completely abrogated by B6 Tregs (Fig. 6A), and the most severe colitis was induced by Sle1a Teff alone (Fig. 6B). A spectrum of severity was observed in mice receiving Sle1a.1 or Sle1a.2 Teffs and Tregs in combination with B6 T cells. The lesser involved colons showed inflammation limited to the crypts (cryptitis) and lamina propria. Increasing inflammation involved the muscularis mucosa and inner submucosal connective tissue stroma. The most severe inflammation involved the muscularis propria, submucosal stroma and serosa. The capillaries and venules in each layer with inflammation showed margination and congestion with mononuclear cells and swelling. Cryptitis was primarily lymphocytic but higher-grade specimens also had focal neutrophilic crypt abscesses. More epithelioid inflammation was also present in the higher-grade lesions in the lamina propria and deeper layers with nodules and occasional multinucleated epithelioid giant cells (Fig. 6B). Increasing inflammation was associated with increasing wall thickness up to three-fold normal (data not shown). Sle1a or Sle1a.1 Tregs did not suppress B6 Teff-induced colitis as well as B6 Tregs (Fig. 6G). Impaired Sle1a.2 Tregs were detected clinically with body weight loss (data not shown), but not by histology (Fig. 6G), indicating that this functional impairment was not as severe as for Sle1a.1. Sle1a.1 Teffs were resistant to suppression by B6 Tregs to a similar level as Sle1a Teffs (Fig. 6G). In contrast, mice reconstituted with B6 Tregs:Sle1a.2 Teffs or B6 Tregs:B6 Teffs had equivalent scores, indicating that Sle1a.2 does not increase Teff inflammatory functions in vivo.

Bottom Line: Sle1a.1, and to a lesser extent Sle1a.2, significantly affected CD4(+) T-cell activation as well as Treg differentiation and function.As the Sle1a.1 and Sle1a.2 intervals contain only 1 and 15 known genes, respectively, this study considerably reduces the number of candidate genes responsible for the production of autoreactive T cells.These results also show that the Sle1 locus is an excellent model for the genetic architecture of lupus, in which a major obligate phenotype results from the coexpression of multiple genetic variants with individual weak effects.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Immunology, and Laboratory Medicine, University of Florida, Gainesville, 32610-0275, USA.

ABSTRACT
The NZM2410-derived Sle1a lupus susceptibility locus induces activated autoreactive CD4(+) T cells and reduces the number and function of Foxp3(+) regulatory T cells (Tregs). In this study, we first showed that Sle1a contributes to autoimmunity by increasing antinuclear antibody production when expressed on either NZB or NZW heterozygous genomes, and by enhancing the chronic graft versus host disease response indicating an expansion of the autoreactive B-cell pool. Screening two non-overlapping recombinants, the Sle1a.1 and Sle1a.2 intervals that cover the entire Sle1a locus, revealed that both Sle1a.1 and Sle1a.2 were necessary for the full Sle1a phenotype. Sle1a.1, and to a lesser extent Sle1a.2, significantly affected CD4(+) T-cell activation as well as Treg differentiation and function. Sle1a.2 also increased the production of autoreactive B cells. As the Sle1a.1 and Sle1a.2 intervals contain only 1 and 15 known genes, respectively, this study considerably reduces the number of candidate genes responsible for the production of autoreactive T cells. These results also show that the Sle1 locus is an excellent model for the genetic architecture of lupus, in which a major obligate phenotype results from the coexpression of multiple genetic variants with individual weak effects.

Show MeSH
Related in: MedlinePlus