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Autoimmunity-associated protein tyrosine phosphatase PEP negatively regulates IFN-α receptor signaling.

Holmes DA, Suto E, Lee WP, Ou Q, Gong Q, Smith HR, Caplazi P, Chan AC - J. Exp. Med. (2015)

Bottom Line: Pep(-/-) hematopoietic progenitors demonstrate increased IFNAR signaling, increased IFN-inducible gene expression, and enhanced proliferation and activation compared to Pep(+/+) progenitors in response to IFN-α.In addition, Pep(-/-) mice treated with IFN-α display a profound defect in hematopoiesis, resulting in anemia, thrombocytopenia, and neutropenia when compared to IFN-α-treated Pep(+/+) mice.As SLE patients carrying the PTPN22(C1858T) risk variant have higher serum IFN-α activity, these data provide a molecular basis for how type I IFNs and PTPN22 may cooperate to contribute to lupus-associated cytopenias.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Immunology, Department of Translational Immunology, and Department of Pathology, Genentech, Inc., South San Francisco, CA 94080.

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Pep−/− mice are susceptible to an IFN-α–mediated lupus-like disease. (A) Pep+/+ (red) and Pep−/− (blue) mice were injected i.v. with adenovirus expressing IFN-α5 (Ad5-IFN-α5, 109 PFU/mouse). Mortality was observed for 60 d after infection. Survival curves represent a total of 25 mice per genotype from 3 independent experiments. (B) Protein levels in the urine were measured in Ad5-IFN-α5 (109 PFU/mouse)–infected Pep+/+ (open circles) and Pep−/− (grey squares) mice 24 d after infection. Data are cumulative of 3 independent experiments representing a total of 11 Pep+/+ or 13 Pep−/− mice. (C) Serum from Ad5-IFN-α5 (109 PFU/mouse)–infected Pep+/+ (open circles) and Pep−/− (grey squares) mice was analyzed at the indicated times for antinuclear IgG antibodies by ELISA. n = 3–5 mice/group and data shown are representative of 3 independent experiment. (D) Kidney sections from Pep+/+ (top) and Pep−/−>(bottom) mice infected with Ad5-IFN-α5 (109 PFU/mouse) 24 d after infection were stained with PAS. Arrows indicate capillary basement membranes and mesangial matrix. Bar, 20 µm. Images are representative of 12–20 sections per genotype from 2 independent experiments. (E–G) Blood neutrophil (E), RBC (F), and PLT (G) numbers from Pep+/+ (open circles) and Pep−/− (grey squares) were analyzed in Ad5-LacZ (109 PFU/mouse) or Ad5-IFN-α5 (109 PFU/mouse) at 14 (E) or 28 d (F–G) after infection. (E) n = 4 mice/group and data shown are representative of 3 independent experiments; (F–G) n = 5 mice/group in LacZ-treated cohort and data shown are representative of 3 independent experiments. n = 25 Pep+/+ and 8 Pep−/− mice in the IFN-α5 cohorts. Data shown are cumulative of three independent experiments. (H and I) Splenic neutrophil (H) and monocyte (I) numbers in Pep+/+ (open circles) and Pep−/− (grey squares) mice were analyzed in control Ad5-LacZ (109 PFU/mouse) or Ad5-IFN-α5 (109 PFU/mouse) 14 d after infection. Data are representative of three independent experiments (n = 4 mice/group). (J) Blood lymphocyte numbers from Pep+/+ (open circles) and Pep−/− (grey squares) mice were analyzed in Ad5-LacZ (109 PFU/mouse) or Ad5-IFN-α5 (109 PFU/mouse) 28 d after infection. n = 4 mice/group in the LacZ-treated cohort; n = 9 Pep+/+ and 5 Pep−/− mice in the IFN-α5–treated cohort. Data shown are representative of three independent experiments. (K) Serum BAFF/BLys levels in Pep+/+ (open circles) and Pep−/− (grey squares) mice were analyzed in control Ad5-LacZ (109 PFU/mouse) or Ad5-IFN-α5 (109 PFU/mouse) 28 d after infection by ELISA. n > 13 mice/group and are cumulative of at least 3 independent experiments. Values in all graphs represent means ± SEM. Statistical analyses were done by two-tailed paired Student’s t test; *, P < 0.05; **, P < 0.01; ***, P < 0.001.
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fig1: Pep−/− mice are susceptible to an IFN-α–mediated lupus-like disease. (A) Pep+/+ (red) and Pep−/− (blue) mice were injected i.v. with adenovirus expressing IFN-α5 (Ad5-IFN-α5, 109 PFU/mouse). Mortality was observed for 60 d after infection. Survival curves represent a total of 25 mice per genotype from 3 independent experiments. (B) Protein levels in the urine were measured in Ad5-IFN-α5 (109 PFU/mouse)–infected Pep+/+ (open circles) and Pep−/− (grey squares) mice 24 d after infection. Data are cumulative of 3 independent experiments representing a total of 11 Pep+/+ or 13 Pep−/− mice. (C) Serum from Ad5-IFN-α5 (109 PFU/mouse)–infected Pep+/+ (open circles) and Pep−/− (grey squares) mice was analyzed at the indicated times for antinuclear IgG antibodies by ELISA. n = 3–5 mice/group and data shown are representative of 3 independent experiment. (D) Kidney sections from Pep+/+ (top) and Pep−/−>(bottom) mice infected with Ad5-IFN-α5 (109 PFU/mouse) 24 d after infection were stained with PAS. Arrows indicate capillary basement membranes and mesangial matrix. Bar, 20 µm. Images are representative of 12–20 sections per genotype from 2 independent experiments. (E–G) Blood neutrophil (E), RBC (F), and PLT (G) numbers from Pep+/+ (open circles) and Pep−/− (grey squares) were analyzed in Ad5-LacZ (109 PFU/mouse) or Ad5-IFN-α5 (109 PFU/mouse) at 14 (E) or 28 d (F–G) after infection. (E) n = 4 mice/group and data shown are representative of 3 independent experiments; (F–G) n = 5 mice/group in LacZ-treated cohort and data shown are representative of 3 independent experiments. n = 25 Pep+/+ and 8 Pep−/− mice in the IFN-α5 cohorts. Data shown are cumulative of three independent experiments. (H and I) Splenic neutrophil (H) and monocyte (I) numbers in Pep+/+ (open circles) and Pep−/− (grey squares) mice were analyzed in control Ad5-LacZ (109 PFU/mouse) or Ad5-IFN-α5 (109 PFU/mouse) 14 d after infection. Data are representative of three independent experiments (n = 4 mice/group). (J) Blood lymphocyte numbers from Pep+/+ (open circles) and Pep−/− (grey squares) mice were analyzed in Ad5-LacZ (109 PFU/mouse) or Ad5-IFN-α5 (109 PFU/mouse) 28 d after infection. n = 4 mice/group in the LacZ-treated cohort; n = 9 Pep+/+ and 5 Pep−/− mice in the IFN-α5–treated cohort. Data shown are representative of three independent experiments. (K) Serum BAFF/BLys levels in Pep+/+ (open circles) and Pep−/− (grey squares) mice were analyzed in control Ad5-LacZ (109 PFU/mouse) or Ad5-IFN-α5 (109 PFU/mouse) 28 d after infection by ELISA. n > 13 mice/group and are cumulative of at least 3 independent experiments. Values in all graphs represent means ± SEM. Statistical analyses were done by two-tailed paired Student’s t test; *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Mentions: Given the genetic links implicating both IFN-α and PTPN22 in human SLE, we tested whether PEP deficiency may affect the ability of IFN-α to induce SLE. Systemic administration of IFN-α5 (109 PFU/mouse) by adenoviral delivery resulted in 40% mortality in Pep+/+ mice (C57BL/6 background) 2 mo after IFN-α administration (Fig. 1 A). In contrast, IFN-α (109 PFU/mouse) treated Pep−/− mice (C57BL/6) demonstrated 40% mortality by 2 wk and ∼90% mortality by 2 mo. Corresponding with increased mortality, IFN-α5–treated Pep−/− mice developed increased proteinuria, a mild glomerulopathy, antinuclear autoantibodies. and cytopenias when compared to IFN-α5–treated Pep+/+ mice (Fig. 1, B–G). Affected glomeruli of IFN-α5–treated Pep−/− mice have segmental or global thickening of capillary basement membranes and increased mesangial matrix (Fig. 1 D, arrows), resulting in markedly narrowed capillary profiles. Using an arbitrary scoring system (described in Materials and methods), the pathology score for IFN-α5–treated Pep+/+ mice was 0.0 + 0, whereas IFN-α5–treated Pep−/− mice was 0.67 + 0.2 (P = 0.03). Cytopenias, including peripheral neutropenia, anemia, and thrombocytopenia, as well as reductions in splenic neutrophils and monocytes, were also more severe in IFN-α5–treated Pep−/− mice (Fig. 1, E–I). In contrast, whereas IFN-α5 reduced peripheral lymphocytes, there were no differences between IFN-α5–treated Pep+/+ and Pep−/− mice (Fig. 1 J). As IFN-α increased serum BAFF/BLys (Mathian et al., 2005), a key pathogenic factor in SLE, BAFF/BLys levels were elevated in IFN-α5–treated mice compared to LacZ controls, but no differences were observed between IFN-α5–treated Pep+/+ and Pep−/− mice (Fig. 1 K). Together, these data suggest that PEP deficiency accelerates IFN-α5–induced lupus-like disease with increased mortality, proteinuria, glomerulopathy, autoantibody production, and cytopenias.


Autoimmunity-associated protein tyrosine phosphatase PEP negatively regulates IFN-α receptor signaling.

Holmes DA, Suto E, Lee WP, Ou Q, Gong Q, Smith HR, Caplazi P, Chan AC - J. Exp. Med. (2015)

Pep−/− mice are susceptible to an IFN-α–mediated lupus-like disease. (A) Pep+/+ (red) and Pep−/− (blue) mice were injected i.v. with adenovirus expressing IFN-α5 (Ad5-IFN-α5, 109 PFU/mouse). Mortality was observed for 60 d after infection. Survival curves represent a total of 25 mice per genotype from 3 independent experiments. (B) Protein levels in the urine were measured in Ad5-IFN-α5 (109 PFU/mouse)–infected Pep+/+ (open circles) and Pep−/− (grey squares) mice 24 d after infection. Data are cumulative of 3 independent experiments representing a total of 11 Pep+/+ or 13 Pep−/− mice. (C) Serum from Ad5-IFN-α5 (109 PFU/mouse)–infected Pep+/+ (open circles) and Pep−/− (grey squares) mice was analyzed at the indicated times for antinuclear IgG antibodies by ELISA. n = 3–5 mice/group and data shown are representative of 3 independent experiment. (D) Kidney sections from Pep+/+ (top) and Pep−/−>(bottom) mice infected with Ad5-IFN-α5 (109 PFU/mouse) 24 d after infection were stained with PAS. Arrows indicate capillary basement membranes and mesangial matrix. Bar, 20 µm. Images are representative of 12–20 sections per genotype from 2 independent experiments. (E–G) Blood neutrophil (E), RBC (F), and PLT (G) numbers from Pep+/+ (open circles) and Pep−/− (grey squares) were analyzed in Ad5-LacZ (109 PFU/mouse) or Ad5-IFN-α5 (109 PFU/mouse) at 14 (E) or 28 d (F–G) after infection. (E) n = 4 mice/group and data shown are representative of 3 independent experiments; (F–G) n = 5 mice/group in LacZ-treated cohort and data shown are representative of 3 independent experiments. n = 25 Pep+/+ and 8 Pep−/− mice in the IFN-α5 cohorts. Data shown are cumulative of three independent experiments. (H and I) Splenic neutrophil (H) and monocyte (I) numbers in Pep+/+ (open circles) and Pep−/− (grey squares) mice were analyzed in control Ad5-LacZ (109 PFU/mouse) or Ad5-IFN-α5 (109 PFU/mouse) 14 d after infection. Data are representative of three independent experiments (n = 4 mice/group). (J) Blood lymphocyte numbers from Pep+/+ (open circles) and Pep−/− (grey squares) mice were analyzed in Ad5-LacZ (109 PFU/mouse) or Ad5-IFN-α5 (109 PFU/mouse) 28 d after infection. n = 4 mice/group in the LacZ-treated cohort; n = 9 Pep+/+ and 5 Pep−/− mice in the IFN-α5–treated cohort. Data shown are representative of three independent experiments. (K) Serum BAFF/BLys levels in Pep+/+ (open circles) and Pep−/− (grey squares) mice were analyzed in control Ad5-LacZ (109 PFU/mouse) or Ad5-IFN-α5 (109 PFU/mouse) 28 d after infection by ELISA. n > 13 mice/group and are cumulative of at least 3 independent experiments. Values in all graphs represent means ± SEM. Statistical analyses were done by two-tailed paired Student’s t test; *, P < 0.05; **, P < 0.01; ***, P < 0.001.
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fig1: Pep−/− mice are susceptible to an IFN-α–mediated lupus-like disease. (A) Pep+/+ (red) and Pep−/− (blue) mice were injected i.v. with adenovirus expressing IFN-α5 (Ad5-IFN-α5, 109 PFU/mouse). Mortality was observed for 60 d after infection. Survival curves represent a total of 25 mice per genotype from 3 independent experiments. (B) Protein levels in the urine were measured in Ad5-IFN-α5 (109 PFU/mouse)–infected Pep+/+ (open circles) and Pep−/− (grey squares) mice 24 d after infection. Data are cumulative of 3 independent experiments representing a total of 11 Pep+/+ or 13 Pep−/− mice. (C) Serum from Ad5-IFN-α5 (109 PFU/mouse)–infected Pep+/+ (open circles) and Pep−/− (grey squares) mice was analyzed at the indicated times for antinuclear IgG antibodies by ELISA. n = 3–5 mice/group and data shown are representative of 3 independent experiment. (D) Kidney sections from Pep+/+ (top) and Pep−/−>(bottom) mice infected with Ad5-IFN-α5 (109 PFU/mouse) 24 d after infection were stained with PAS. Arrows indicate capillary basement membranes and mesangial matrix. Bar, 20 µm. Images are representative of 12–20 sections per genotype from 2 independent experiments. (E–G) Blood neutrophil (E), RBC (F), and PLT (G) numbers from Pep+/+ (open circles) and Pep−/− (grey squares) were analyzed in Ad5-LacZ (109 PFU/mouse) or Ad5-IFN-α5 (109 PFU/mouse) at 14 (E) or 28 d (F–G) after infection. (E) n = 4 mice/group and data shown are representative of 3 independent experiments; (F–G) n = 5 mice/group in LacZ-treated cohort and data shown are representative of 3 independent experiments. n = 25 Pep+/+ and 8 Pep−/− mice in the IFN-α5 cohorts. Data shown are cumulative of three independent experiments. (H and I) Splenic neutrophil (H) and monocyte (I) numbers in Pep+/+ (open circles) and Pep−/− (grey squares) mice were analyzed in control Ad5-LacZ (109 PFU/mouse) or Ad5-IFN-α5 (109 PFU/mouse) 14 d after infection. Data are representative of three independent experiments (n = 4 mice/group). (J) Blood lymphocyte numbers from Pep+/+ (open circles) and Pep−/− (grey squares) mice were analyzed in Ad5-LacZ (109 PFU/mouse) or Ad5-IFN-α5 (109 PFU/mouse) 28 d after infection. n = 4 mice/group in the LacZ-treated cohort; n = 9 Pep+/+ and 5 Pep−/− mice in the IFN-α5–treated cohort. Data shown are representative of three independent experiments. (K) Serum BAFF/BLys levels in Pep+/+ (open circles) and Pep−/− (grey squares) mice were analyzed in control Ad5-LacZ (109 PFU/mouse) or Ad5-IFN-α5 (109 PFU/mouse) 28 d after infection by ELISA. n > 13 mice/group and are cumulative of at least 3 independent experiments. Values in all graphs represent means ± SEM. Statistical analyses were done by two-tailed paired Student’s t test; *, P < 0.05; **, P < 0.01; ***, P < 0.001.
Mentions: Given the genetic links implicating both IFN-α and PTPN22 in human SLE, we tested whether PEP deficiency may affect the ability of IFN-α to induce SLE. Systemic administration of IFN-α5 (109 PFU/mouse) by adenoviral delivery resulted in 40% mortality in Pep+/+ mice (C57BL/6 background) 2 mo after IFN-α administration (Fig. 1 A). In contrast, IFN-α (109 PFU/mouse) treated Pep−/− mice (C57BL/6) demonstrated 40% mortality by 2 wk and ∼90% mortality by 2 mo. Corresponding with increased mortality, IFN-α5–treated Pep−/− mice developed increased proteinuria, a mild glomerulopathy, antinuclear autoantibodies. and cytopenias when compared to IFN-α5–treated Pep+/+ mice (Fig. 1, B–G). Affected glomeruli of IFN-α5–treated Pep−/− mice have segmental or global thickening of capillary basement membranes and increased mesangial matrix (Fig. 1 D, arrows), resulting in markedly narrowed capillary profiles. Using an arbitrary scoring system (described in Materials and methods), the pathology score for IFN-α5–treated Pep+/+ mice was 0.0 + 0, whereas IFN-α5–treated Pep−/− mice was 0.67 + 0.2 (P = 0.03). Cytopenias, including peripheral neutropenia, anemia, and thrombocytopenia, as well as reductions in splenic neutrophils and monocytes, were also more severe in IFN-α5–treated Pep−/− mice (Fig. 1, E–I). In contrast, whereas IFN-α5 reduced peripheral lymphocytes, there were no differences between IFN-α5–treated Pep+/+ and Pep−/− mice (Fig. 1 J). As IFN-α increased serum BAFF/BLys (Mathian et al., 2005), a key pathogenic factor in SLE, BAFF/BLys levels were elevated in IFN-α5–treated mice compared to LacZ controls, but no differences were observed between IFN-α5–treated Pep+/+ and Pep−/− mice (Fig. 1 K). Together, these data suggest that PEP deficiency accelerates IFN-α5–induced lupus-like disease with increased mortality, proteinuria, glomerulopathy, autoantibody production, and cytopenias.

Bottom Line: Pep(-/-) hematopoietic progenitors demonstrate increased IFNAR signaling, increased IFN-inducible gene expression, and enhanced proliferation and activation compared to Pep(+/+) progenitors in response to IFN-α.In addition, Pep(-/-) mice treated with IFN-α display a profound defect in hematopoiesis, resulting in anemia, thrombocytopenia, and neutropenia when compared to IFN-α-treated Pep(+/+) mice.As SLE patients carrying the PTPN22(C1858T) risk variant have higher serum IFN-α activity, these data provide a molecular basis for how type I IFNs and PTPN22 may cooperate to contribute to lupus-associated cytopenias.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Immunology, Department of Translational Immunology, and Department of Pathology, Genentech, Inc., South San Francisco, CA 94080.

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Related in: MedlinePlus